Optimizing plant transporter expression in Xenopus oocytes

BACKGROUND: Rapid improvements in DNA synthesis technology are revolutionizing gene cloning and the characterization of their encoded proteins. Xenopus laevis oocytes are a commonly used heterologous system for the expression and functional characterization of membrane proteins. For many plant proteins, particularly transporters, low levels of expression can limit functional activity in these cells making it difficult to characterize the protein. Improvements in synthetic DNA technology now make it quick, easy and relatively cheap to optimize the codon usage of plant cDNAs for Xenopus. We have tested if this optimization process can improve the functional activity of a two-component plant nitrate transporter assayed in oocytes. RESULTS: We used the generally available software (http://www.kazusa.or.jp/codon/; http://genomes.urv.es/OPTIMIZER/) to predict a DNA sequence for the plant gene that is better suited for Xenopus laevis. Rice OsNAR2.1 and OsNRT2.3a DNA optimized sequences were commercially synthesized for Xenopus expression. The template DNA was used to synthesize cRNA using a commercially available kit. Oocytes were injected with cRNA mixture of optimized and original OsNAR2.1 and OsNRT2.3a. Oocytes injected with cRNA obtained from using the optimized DNA template could accumulate significantly more NO(3)(-) than the original genes after 16 h incubation in 0.5 mM Na(15)NO(3). Two-electrode voltage clamp analysis of the oocytes confirmed that the codon optimized template resulted in significantly larger currents when compared with the original rice cDNA. CONCLUSION: The functional activity of a rice high affinity nitrate transporter in oocytes was improved by DNA codon optimization of the genes. This methodology offers the prospect for improved expression and better subsequent functional characterization of plant proteins in the Xenopus oocyte system

Degradation of Toxic Organic Contaminants by Graphene Cathode in an Electro‐Fenton System

A novel composite electrode was constructed by pressing graphene and CuO, using a cathode in an electro‐Fenton (EF) system. Cyclic voltammetry, charge/discharge curve and electrochemical impedance spectroscopy (EIS) were used to characterize the composite electrode. The degradation of a toxic organic contaminant, Terramycin, by EF system was studied in an undivided electrolysis cell. The possible degradation products of Terramycin were studied by a Fourier transform‐infrared spectrum, and the findings showed that the structure of Terramycin was damaged. The variations of hydrogen peroxide and the relative content of hydroxyl radical (.OH) during the degradation process were traced by enzyme catalysis method and fluorescence spectrometry. The results showed that the electro‐catalytic degradation of Terramycin occurred by an ·OH radical mechanism. More importantly, this as‐prepared cathode was very stable and could be reused without any catalytic activity decrease, suggesting its potential application in the wastewater treatment

Prevention of post-surgical abdominal adhesions by a novel biodegradable thermosensitive PECE hydrogel.

<p>Abstract</p> <p>Background</p> <p>Post-operative peritoneal adhesions are common and serious complications for modern medicine. We aim to prevent post-surgical adhesions using biodegradable and thermosensitive poly(ethylene glycol)-poly(ε-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) hydrogel. In this work, we investigated the effect of PECE hydrogel on preventing post-surgical abdominal adhesions in mouse and rat models.</p> <p>Results</p> <p>The PECE hydrogel in sol state could be transformed into gel in less than 20 s at 37°C. In addition, the PECE hydrogel could be easily adhered to the damaged peritoneal surfaces, and be gradually degraded and absorbed by the body within 14 days along with the healing of peritoneal wounds. A notable efficacy of the PECE hydrogel in preventing peritoneal adhesions was demonstrated in the animal models. In contrast, all untreated animals developed adhesions requiring sharp dissection. Furthermore, no significant histopathological changes were observed in main organs of the hydrogel-treated animals.</p> <p>Conclusion</p> <p>Our results suggested that the thermosensitive PECE hydrogel was an effective, safe, and convenient agent on preventing post-surgical intro-abdominal adhesions.</p

THE EFFECTS OF MAGGOT SECRETIONS ON THE INFLAMMATORY CYTOKINES IN SERUM OF TRAUMATIC RATS

The objective of this study was to analyse the changes of inflammatory cytokines level in traumatic rat serum after the maggot secretions intervention. Acute traumatic rat were randomly divided into three groups that included maggot secretions group, negative group and the control group. TNF-α, IL-6, SOD, LPO levels were measured by radioimmunoassay. The experimental results showed that TNF-α, IL-6, and SOD levels in the model group were significantly increased; LPO level was decreased and showed significant differences. Thus, the content of inflammatory cytokines in acute skin wounds could be reduced by maggot secretions, which play a role in enhancing wound healing

Observation of the Anomalous Hall Effect in a Collinear Antiferromagnet

Time-reversal symmetry breaking is the basic physics concept underpinning many magnetic topological phenomena such as the anomalous Hall effect (AHE) and its quantized variant. The AHE has been primarily accompanied by a ferromagnetic dipole moment, which hinders the topological quantum states and limits data density in memory devices, or by a delicate noncollinear magnetic order with strong spin decoherence, both limiting their applicability. A potential breakthrough is the recent theoretical prediction of the AHE arising from collinear antiferromagnetism in an anisotropic crystal environment. This new mechanism does not require magnetic dipolar or noncollinear fields. However, it has not been experimentally observed to date. Here we demonstrate this unconventional mechanism by measuring the AHE in an epilayer of a rutile collinear antiferromagnet RuO$_2$. The observed anomalous Hall conductivity is large, exceeding 300 S/cm, and is in agreement with the Berry phase topological transport contribution. Our results open a new unexplored chapter of time-reversal symmetry breaking phenomena in the abundant class of collinear antiferromagnetic materials.Comment: 33 pages, 14 figures, 2 table

Metabolomics analysis of herb-partitioned moxibustion treatment on rats with diarrhea-predominant irritable bowel syndrome

Background: Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder, which is commonly treated with antidiarrhoeal, antispasmodics, serotonergic agents or laxative agents. These treatments provide relief for IBS symptoms but may also lead to undesired side effects. Previously, herb-partitioned moxibustion (HPM) treatment has been demonstrated to be effective in ameliorating symptoms of IBS. However, the underlying mechanism of this beneficial treatment is yet to be established. The aim of the current study was to systematically assess the metabolic alterations in response to diarrhea-predominant IBS (IBS-D) and therapeutic effect of HPM. Methods: Proton nuclear magnetic resonance spectroscopy (1H NMR)-based metabolomics approach was used to investigate fecal and serum metabolome of rat model of IBS-D with and without HPM treatment. Results: The current results showed that IBS-induced metabolic alterations in fecal and serum sample include higher level of threonine and UDP-glucose together with lower levels of aspartate, ornithine, leucine, isoleucine, proline, 2-hydroxy butyrate, valine, lactate, ethanol, arginine, 2-oxoisovalerate and bile acids. These altered metabolites potentially involve in impaired gut secretory immune system and intestinal inflammation, malabsorption of nutrients, and disordered metabolism of bile acids. Notably, the HPM treatment was found able to normalize the Bristol stool forms scale scores, fecal water content, plasma endotoxin level, and a number of IBS-induced metabolic changes. Conclusions: These findings may provide useful insight into the molecular basis of IBS and mechanism of the HPM intervention

Biochar-based fertilizer: Supercharging root membrane potential and biomass yield of rice

Biochar-based compound fertilizers (BCF) and amendments have proven to enhance crop yields and modify soil properties (pH, nutrients, organic matter, structure etc.) and are now in commercial production in China. While there is a good understanding of the changes in soil properties following biochar addition, the interactions within the rhizosphere remain largely unstudied, with benefits to yield observed beyond the changes in soil properties alone. We investigated the rhizosphere interactions following the addition of an activated wheat straw BCF at an application rates of 0.25% (g·g−1 soil), which could potentially explain the increase of plant biomass (by 67%), herbage N (by 40%) and P (by 46%) uptake in the rice plants grown in the BCF-treated soil, compared to the rice plants grown in the soil with conventional fertilizer alone. Examination of the roots revealed that micron and submicron-sized biochar were embedded in the plaque layer. BCF increased soil Eh by 85 mV and increased the potential difference between the rhizosphere soil and the root membrane by 65 mV. This increased potential difference lowered the free energy required for root nutrient accumulation, potentially explaining greater plant nutrient content and biomass. We also demonstrate an increased abundance of plant-growth promoting bacteria and fungi in the rhizosphere. We suggest that the redox properties of the biochar cause major changes in electron status of rhizosphere soils that drive the observed agronomic benefits

Transcriptome Analysis on Maternal Separation Rats With Depression-Related Manifestations Ameliorated by Electroacupuncture

Maternal separation (MS), a stressful event in early life, has been linked to neuropsychiatric disorders later in life, especially depression. In this study we investigated whether treatment with electroacupuncture (EA) could ameliorate depression-related manifestations in adult animals that had adverse early life experiences. We demonstrated depression-like behavior deficiencies in a sucrose preference test and a forced swimming test in a rat model with neonatal MS. Repeated EA treatment at the acupoints Baihui (GV20) and Yintang (GV29) during adulthood was shown to be remarkably attenuated above behavioral deficits. Using unbiased genome-wide RNA sequencing to investigate alterations in the transcriptome of the prefrontal cortex (PFC), we explored the altered gene sets involved in circadian rhythm and neurotransmitter transporter activity in MS rats, and their expression tended to be reversed after EA treatment. In addition, we analyzed the interaction network of differentiated lncRNA– or circRNA–miRNA–mRNA by using the principle of competitive endogenous RNA (ceRNA). These results suggest that EA at GV20 and GV29 ameliorates depression-related manifestations by regulating the expression of multiple genes

Publisher Correction: An anomalous Hall effect in altermagnetic ruthenium dioxide

In the version of this article initially published, square brackets and parentheses were incorrect in Fig. 1g and throughout Fig. 2 (excepting lower labels in Fig. 2d–f). Further, in the second paragraph of the “Consistency with theoretical prediction” subsection of the main article, in the text now reading “the reorientation-field scale, namely, HC = (H2 AE − H2 d) /Hd,” the term “H2 AE” wasn’t shown as squared. The changes have been made in the HTML and PDF versions of the article