Evaluation of wastewater nitrogen transformation in a natural wetland (Ulaanbaatar, Mongolia) using dual-isotope analysis of nitrate

The Tuul River, which provides water for the daily needs of many residents of Ulaanbaatar, Mongolia, has been increasingly polluted by wastewater from the city's sewage treatment plant. Information on water movement and the transformation of water-borne materials is required to alleviate the deterioration of water quality. We conducted a synoptic survey of general water movement, water quality including inorganic nitrogen concentrations, and isotopic composition of nitrogen (δ¹⁵N-NO₃⁻, δ¹⁸O-NO₃⁻, and δ¹⁵N-NH₄⁺) and water (δ¹⁸O-H₂O) in a wetland area that receives wastewater before it enters the Tuul River. We sampled surface water, groundwater, and spring water along the two major water routes in the wetland that flow from the drain of the sewage treatment plant to the Tuul River: a continuous tributary and a discontinuous tributary. The continuous tributary had high ammonium (NH₄⁺) concentrations and nearly stable δ¹⁵N-NH₄⁺, δ¹⁵N-NO₃⁻, and δ¹⁸O-NO₃⁻ concentrations throughout its length, indicating that nitrogen transformation (i.e., nitrification and denitrification) during transit was small. In contrast, NH₄⁺ concentrations decreased along the discontinuous tributary and nitrate (NO₃⁻) concentrations were low at many points. Values of δ¹⁵N-NH₄⁺, δ¹⁵N-NO₃⁻, and δ¹⁸O-NO₃⁻ increased with flow along the discontinuous route. Our results indicate that nitrification and denitrification contribute to nitrogen removal in the wetland area along the discontinuous tributary with slow water transport. Differences in hydrological pathways and the velocity of wastewater transport through the wetland area greatly affect the extent of nitrogen removal.     

Acid tolerance and gad mRNA levels of Escherichia coli O157:H7 grown in foods

We examined the acid tolerance and gad mRNA levels of Escherichia coli O157:H7 (three strains) and nonpathogenic E. coli (strains K12, W1485, and B) grown in foods. The E. coli cells (approximately 30,000 cells) were inoculated on the surface of 10 g of solid food samples (asparagus, broccoli, carrot, celery, cucumber, eggplant, ginger, green pepper, onion, potato, radish, tomato and beef) and in 10 ml of cow's milk, cultured statically at 10-25 degrees C for 1-14 days, and subjected to an acid challenge at 37 degrees C for 1 h in LB medium (pH 3.0). When grown at 20 and 25 degrees C in all foods, except for tomato and ginger, the strains showed a stationary-phase specific acid tolerance. The acid tolerance of the O157 strains changed depending on the types of foods (3-10% survival), but was clearly lower than that of the cells grown in EC medium (more than 90% survival). Tomato and ginger induced relatively high acid tolerances (10-30% survival) in the O157 strains irrespective of the growth phase, probably because of their acidity. No remarkable difference was observed in the acid tolerance between the O157 and nonpathogenic strains grown in all foods. When grown at 10 and 15 degrees C in the foods and EC medium, none of the strains showed the stationary-phase specific acid tolerance. In beef, broccoli, celery, potato and radish, the acid tolerance showed a tendency to decrease with the prolonged cultivation time. In other foods, the acid tolerance was almost constant (about 0.1% survival) irrespective of the growth stage. The mRNA level of glutamate decarboxylase genes (gadA and gadB) correlated to the acid tolerance level when the E. coli cells were grown at 25 degrees C, but was very low even in the stationary phase when the E. coli cells were grown at 15 degrees C or below.     

A ROCK Inhibitor Promotes Graft Survival during Transplantation of iPS-Cell-Derived Retinal Cells

Currently, retinal pigment epithelium (RPE) transplantation includes sheet and single-cell transplantation, the latter of which includes cell death and may be highly immunogenic, and there are some issues to be improved in single-cell transplantation. Y-27632 is an inhibitor of Rho-associated protein kinase (ROCK), the downstream kinase of Rho. We herein investigated the effect of Y-27632 in vitro on retinal pigment epithelium derived from induced pluripotent stem cells (iPS-RPE cells), and also its effects in vivo on the transplantation of iPS-RPE cell suspensions. As a result, the addition of Y-27632 in vitro showed suppression of apoptosis, promotion of cell adhesion, and higher proliferation and pigmentation of iPS-RPE cells. Y-27632 also increased the viability of the transplant without showing obvious retinal toxicity in human iPS-RPE transplantation into monkey subretinal space in vivo. Therefore, it is possible that ROCK inhibitors can improve the engraftment of iPS-RPE cell suspensions after transplantation.     

Drusenoid Pigment Epithelial Detachment: Genetic and Clinical Characteristics

Few studies report drusenoid pigment epithelial detachment (DPED) in Asians. In this multicenter study, we report the clinical and genetic characteristics of 76 patients with DPED, and, for comparison, 861 patients with exudative age-related macular degeneration (AMD) were included. On the initial presentation, the mean best-corrected visual acuity was 0.087 ± 0.17 (logMAR unit), and mean DPED height and width were 210 ± 132 and 1633 ± 1114 µm, respectively. Fifty-one (67%) patients showed macular neovascularization in the contralateral eye. The risk allele frequency of both ARMS2 A69S and CFH I62V was significantly higher in DPED than in typical AMD and polypoidal choroidal vasculopathy (PCV) (ARMS2 A69S risk allele frequency: DPED 77% vs. typical AMD 66% vs. PCV 57%, CFH I62V risk allele frequency: DPED 87% vs. typical AMD 73% vs. PCV 73%), although the risk allele frequency of both genes was similar between the DPED group and retinal angiomatous proliferation (RAP) group (ARMS2 A69S: p = 0.32, CFH I62V, p = 0.11). The prevalence of reticular pseudodrusen (RPD) was highest in RAP (60%), followed by DPED (22%), typical AMD (20%), and PCV (2%). Although the prevalence of RPD differs between DPED and RAP, these entities share a similar genetic background in terms of ARMS2 and CFH genes.     

Antiatherogenic effect of guava leaf extracts inhibiting leucocyte-type 12-lipoxygenase activity

Oxidative modification of low density lipoprotein (LDL) is one of the critical steps for the development of atherosclerosis and leucocyte-type 12-lipoxygenase, highly expressed in macrophages, has been suggested to play an essential role in this process. In the present study, we show that guava leaf extracts inhibited, not only the leucocyte-type 12-lipoxygenase activity, but also LDL oxidation, mediated by the enzyme-overexpressing macrophage-like J774A.1 cells. Oral administration of guava leaf extracts to apoE-knockout mice at 100 mg of dry extracts/kg of body weight, once a day for 16 weeks, significantly reduced the area of atherogenic lesions developed in the aorta and aortic sinus. The major components inhibiting leucocyte-type 12-lipoxygenase contained in guava leaf extracts were identified as ethyl gallate and quercetin. The inhibitory effects of guava leaf extracts on the leucocyte-type 12-lipoxygenase activity, as well as on cell-mediated LDL oxidation, might be involved in the antiatherogenic effect of the extracts.     

Long Non-Coding RNA CRNDE Is Involved in Resistance to EGFR Tyrosine Kinase Inhibitor in EGFR-Mutant Lung Cancer via eIF4A3/MUC1/EGFR Signaling

(1) Background: Acquired resistance to epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) is an intractable problem for many clinical oncologists. The mechanisms of resistance to EGFR-TKIs are complex. Long non-coding RNAs (lncRNAs) may play an important role in cancer development and metastasis. However, the biological process between lncRNAs and drug resistance to EGFR-mutated lung cancer remains largely unknown. (2) Methods: Osimertinib- and afatinib-resistant EGFR-mutated lung cancer cells were established using a stepwise method. A microarray analysis of non-coding and coding RNAs was performed using parental and resistant EGFR-mutant non-small cell lung cancer (NSCLC) cells and evaluated by bioinformatics analysis through medical-industrial collaboration. (3) Results: Colorectal neoplasia differentially expressed (CRNDE) and DiGeorge syndrome critical region gene 5 (DGCR5) lncRNAs were highly expressed in EGFR-TKI-resistant cells by microarray analysis. RNA-protein binding analysis revealed eukaryotic translation initiation factor 4A3 (eIF4A3) bound in an overlapping manner to CRNDE and DGCR5. The CRNDE downregulates the expression of eIF4A3, mucin 1 (MUC1), and phospho-EGFR. Inhibition of CRNDE activated the eIF4A3/MUC1/EGFR signaling pathway and apoptotic activity, and restored sensitivity to EGFR-TKIs. (4) Conclusions: The results showed that CRNDE is associated with the development of resistance to EGFR-TKIs. CRNDE may be a novel therapeutic target to conquer EGFR-mutant NSCLC.     

Effect of surface oxide layer on deuterium permeation behaviors through a type 316 stainless steel

Effect of surface oxide layer on the hydrogen isotope permeation was studied. Iron oxide was uniformly formed in the oxide layer, although chromium was limited at the interface between the oxide layer and bulk SS-316. The permeation behavior of deuterium for oxidized SS-316 was compared with that for unoxidized SS-316 at temperature range of 333–673 K. The deuterium permeability for the oxidized SS-316 was reduced 1/10–1/20 times as high as that for unoxidized one. However, the activation energy of deuterium permeation as gas form for oxidized SS-316 was almost the same as that for unoxidized SS-316 and was 0.64 eV, which was almost consistent with the sum of activation energies for diffusion and solubility. This fact indicates that the deuterium permeation is diffusion limited. The permeability of deuterium as water form was almost constant even if heating temperature is high, showing that the deuterium was permeated through bulk SS-316 and react with oxygen at the oxide layer as water desorption, which is controlled by the permeation flux of deuterium and oxygen concentration on the surface of oxide layer in downstream side.     

Heteropolyacid-conjugated chitosan matrix for triphase catalyst

H₃PW₁₂O₄₀ (12-phosphotungstic acid; PWA) is efficient catalysts for the selective oxidation of alkanes, aldehydes, and perfluorinated acids. However, PWA molecules are high solubility in water and this property has been making it difficult to utilize as an industrial catalyst. In this study, the water-insoluble PWA-conjugated chitosan matrix was prepared by mixing the aqueous PWA and chitosan solutions. This conjugate matrix was constructed by the electrostatic interaction between amino group of chitosan and negative charge of PWA and showed the thermal stability at <100 °C. In addition, the PWA-conjugated chitosan matrix indicated the property for triphase catalyst as epoxidation of allylic alcohols, such as cinnamyl alcohol, farnesol, and so on. The isolated yield of epoxidation with the conjugate matrix was as same as that of pure PWA. Furthermore, since this conjugate matrix did not dissolve in water, matrices could easily separate from the reaction solvent and recycle more than 3 times. Therefore, the PWA-conjugated chitosan matrices may have a potential to utilize the novel catalyst for the epoxidation of various allylic alcohols.