AtHKT1;1 Mediates Nernstian Sodium Channel Transport Properties in Arabidopsis Root Stelar Cells

The Arabidopsis AtHKT1;1 protein was identified as a sodium (Na+) transporter by heterologous expression in Xenopus laevis oocytes and Saccharomyces cerevisiae. However, direct comparative in vivo electrophysiological analyses of a plant HKT transporter in wild-type and hkt loss-of-function mutants has not yet been reported and it has been recently argued that heterologous expression systems may alter properties of plant transporters, including HKT transporters. In this report, we analyze several key functions of AtHKT1;1-mediated ion currents in their native root stelar cells, including Na+ and K+ conductances, AtHKT1;1-mediated outward currents, and shifts in reversal potentials in the presence of defined intracellular and extracellular salt concentrations. Enhancer trap Arabidopsis plants with GFP-labeled root stelar cells were used to investigate AtHKT1;1-dependent ion transport properties using patch clamp electrophysiology in wild-type and athkt1;1 mutant plants. AtHKT1;1-dependent currents were carried by sodium ions and these currents were not observed in athkt1;1 mutant stelar cells. However, K+ currents in wild-type and athkt1;1 root stelar cell protoplasts were indistinguishable correlating with the Na+ over K+ selectivity of AtHKT1;1-mediated transport. Moreover, AtHKT1;1-mediated currents did not show a strong voltage dependence in vivo. Unexpectedly, removal of extracellular Na+ caused a reduction in AtHKT1;1-mediated outward currents in Columbia root stelar cells and Xenopus oocytes, indicating a role for external Na+ in regulation of AtHKT1;1 activity. Shifting the NaCl gradient in root stelar cells showed a Nernstian shift in the reversal potential providing biophysical evidence for the model that AtHKT1;1 mediates passive Na+ channel transport properties

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Prominent Crystallization Promotion Effect of Montmorillonite on PTT/PC Blends with PTT as the Continuous Phase

To regulate the crystallization of poly(trimethylene terephthalate) (PTT) retarded by melt blending with polycarbonate (PC), the crystallization of the PTT/PC blend was investigated employing nano-montmorillonite (MMT) as a crystallization promoter with PTT as the continuous phase. The results showed that MMT exhibits a significant promoting effect on PTT crystallization; the presence of 1 wt. % MMT shifts the initial and peak crystallization temperatures of the 70/30 PTT/PC blend to ~17 &deg;C and ~32 &deg;C, respectively. Additionally, the full width at half maximum (FWHM) narrows by ~45%, and the &Delta;Hc increases by 3.7 J.g&minus;1. The accelerating effect of MMT is determined by its distribution and dispersion which depends on the shear intensity, mixing mode, and loading. MMT is easier to exfoliate via the two-step method than by the one-step method. The distribution in the PTT phase is enriched along the phase interface forming an MMT layer. This endows sections of the PTT with abundant nuclei and thus crystallization is promoted markedly compared with the one-step method. Moreover, the finer MMT migrates more readily to the interface to cause a much smoother phase interface. However, a secondary crystallization peak appears when the shear force is not sufficient enough to make MMT finely dispersed, in case of the two-step method and the MMT content is increased to 3 wt. %. The mixing temperature shows little effect on the acceleration of MMT on the crystallization of PTT/PC compared with the shear force. Only when MMT did not exfoliate or uncomplete did the presence of epoxy resin help to promote crystallization because of the improved MMT dispersion

Environmental fate of the anti-parasitic ivermectin in an aquatic micro-ecological system after a single oral administration

Background Ivermectin (IVM) has been widely used in the aquaculture industry since its efficacy against parasites. However, the degradation of IVM was very slow in aquatic environments and the environmental fate of IVM in a complete aquatic system was still not clear. Therefore, comparable studies in a complete aquatic system were merited and helped to elucidate the environmental fate and effects of IVM. Methods An aquatic micro-ecological system containing an aquatic environment (water and sediment) and aquatic organisms (invertebrates, aquatic plants and fish) was built to simulate the natural rearing conditions. A single dose of 0.3 mg kg−1 body weight of IVM was given to the fish by oral gavage. Water, sediment, the roots and leaves of the aquatic plants, the soft tissue of the invertebrates and the visceral mass and muscle of fish samples were collected at 0.5 hours, 1 day, 7 days, 15 days, 30 days, 45 days, 60 days and 70 days after the treatment. IVM concentration in each sample was determined using ELISA method. Results IVM was quickly and widely distributed into the whole aquatic system in one day, and then was highly accumulated in organisms resulting in long-term residues. IVM was exchanged multiple times between the different media, which caused continuous fluctuations in the concentration of IVM in the water and sediment. It was worth noting that there was a second peak value of IVM in the fish and invertebrates after 30 days. The environmental fate of the IVM in the aquatic micro-ecological system showed that the drug was transferred from the fish to aquatic plants in the first seven days, and then gathered in the water and sediment, finally accumulating in the invertebrates. Our results indicated that an effective aquatic micro-ecological system was successfully established, and it could be applied to the study the environmental fate of IVM, which will aid the scientific use of this anti-parasitic agent during aquaculture

Heat Transfer Characteristics of High-Temperature Dusty Flue Gas from Industrial Furnaces in a Granular Bed with Buried Tubes

Experimental heat transfer equipment with a buried tube granular bed was set up for waste heat recovery of flue gas. The effects of flue gas inlet temperature (1096.65&ndash;1286.45 K) and cooling water flow rate (2.6&ndash;5.1 m3/h) were studied through experiment and computational fluid dynamics&rsquo; (CFD) method. On the basis of logarithmic mean temperature difference method, the total heat transfer coefficient of the granular bed was used to characterize its heat transfer performance. Experimental results showed that the waste heat recovery rate of the equipment exceeded 72%. An increase in the cooling water flow rate and inlet gas temperature was beneficial to recovering waste heat. The cooling water flow rate increases from 2.6 m3/h to 5.1 m3/h and the recovery rate of waste heat increases by 1.9%. Moreover, the heat transfer coefficient of the granular bed increased by 4.4% and the inlet gas temperature increased from 1096.65 K to 1286.45 K. The recovery rate of waste heat increased by 1.7% and the heat transfer coefficient of the granular bed rose by 26.6%. Therefore, experimental correlations between the total heat transfer coefficient of a granular bed and the cooling water flow rate and inlet temperature of dusty gas were proposed. The CFD method was used to simulate the heat transfer in the granular bed, and the effect of gas temperature on the heat transfer coefficient of granular bed was studied. Results showed that the relative error was less than 2%

Differential Sodium and Potassium Transport Selectivities of the Rice OsHKT2;1 and OsHKT2;2 Transporters in Plant Cells1[C][OA]

Na+ and K+ homeostasis are crucial for plant growth and development. Two HKT transporter/channel classes have been characterized that mediate either Na+ transport or Na+ and K+ transport when expressed in Xenopus laevis oocytes and yeast. However, the Na+/K+ selectivities of the K+-permeable HKT transporters have not yet been studied in plant cells. One study expressing 5′ untranslated region-modified HKT constructs in yeast has questioned the relevance of cation selectivities found in heterologous systems for selectivity predictions in plant cells. Therefore, here we analyze two highly homologous rice (Oryza sativa) HKT transporters in plant cells, OsHKT2;1 and OsHKT2;2, that show differential K+ permeabilities in heterologous systems. Upon stable expression in cultured tobacco (Nicotiana tabacum) Bright-Yellow 2 cells, OsHKT2;1 mediated Na+ uptake, but little Rb+ uptake, consistent with earlier studies and new findings presented here in oocytes. In contrast, OsHKT2;2 mediated Na+-K+ cotransport in plant cells such that extracellular K+ stimulated OsHKT2;2-mediated Na+ influx and vice versa. Furthermore, at millimolar Na+ concentrations, OsHKT2;2 mediated Na+ influx into plant cells without adding extracellular K+. This study shows that the Na+/K+ selectivities of these HKT transporters in plant cells coincide closely with the selectivities in oocytes and yeast. In addition, the presence of external K+ and Ca2+ down-regulated OsHKT2;1-mediated Na+ influx in two plant systems, Bright-Yellow 2 cells and intact rice roots, and also in Xenopus oocytes. Moreover, OsHKT transporter selectivities in plant cells are shown to depend on the imposed cationic conditions, supporting the model that HKT transporters are multi-ion pores