The zinc finger protein DHHC09 S-acylates the kinase STRK1 to regulate H2O2 homeostasis and promote salt tolerance in rice.

Soil salinity results in oxidative stress and heavy losses to crop production. The S-acylated protein SALT TOLERANCE RECEPTOR-LIKE CYTOPLASMIC KINASE 1 (STRK1) phosphorylates and activates CATALASE C (CatC) to improve rice (Oryza sativa L.) salt tolerance, but the molecular mechanism underlying its S-acylation involved in salt signal transduction awaits elucidation. Here, we show that the DHHC-type zinc finger protein DHHC09 S-acylates STRK1 at Cys5, Cys10, and Cys14 and promotes salt and oxidative stress tolerance by enhancing rice H2O2-scavenging capacity. This modification determines STRK1 targeting to the plasma membrane or lipid nanodomains and is required for its function. DHHC09 promotes salt signaling from STRK1 to CatC via transphosphorylation, and its deficiency impairs salt signal transduction. Our findings demonstrate that DHHC09 S-acylates and anchors STRK1 to the plasma membrane to promote salt signaling from STRK1 to CatC, thereby regulating H2O2 homeostasis and improving salt stress tolerance in rice. Moreover, overexpression of DHHC09 in rice mitigates grain yield loss under salt stress. Together, these results shed light on the mechanism underlying the role of S-acylation in RLK/RLCK-mediated salt signal transduction and provide a strategy for breeding highly salt-tolerant rice.

The protein phosphatase PC1 dephosphorylates and deactivates CatC to negatively regulate H2O2 homeostasis and salt tolerance in rice.

Catalase (CAT) is often phosphorylated and activated by protein kinases to maintain hydrogen peroxide (H2O2) homeostasis and protect cells against stresses, but whether and how CAT is switched off by protein phosphatases remains inconclusive. Here, we identified a manganese (Mn2+)-dependent protein phosphatase, which we named PHOSPHATASE OF CATALASE 1 (PC1), from rice (Oryza sativa L.) that negatively regulates salt and oxidative stress tolerance. PC1 specifically dephosphorylates CatC at Ser-9 to inhibit its tetramerization and thus activity in the peroxisome. PC1 overexpressing lines exhibited hypersensitivity to salt and oxidative stresses with a lower phospho-serine level of CATs. Phosphatase activity and seminal root growth assays indicated that PC1 promotes growth and plays a vital role during the transition from salt stress to normal growth conditions. Our findings demonstrate that PC1 acts as a molecular switch to dephosphorylate and deactivate CatC and negatively regulate H2O2 homeostasis and salt tolerance in rice. Moreover, knockout of PC1 not only improved H2O2-scavenging capacity and salt tolerance but also limited rice grain yield loss under salt stress conditions. Together, these results shed light on the mechanisms that switch off CAT and provide a strategy for breeding highly salt-tolerant rice.

Effects of different order spiking on bioavailability and ecological risk of phenanthrene in mangrove sediment-biochar system.

Biochar shows unique advantage in decreasing the bioavailability of phenanthrene and has huge potential into the in-situ remediation of contaminated sediment. The different order spiking influences the bioavailability and ecological risk of phenanthrene, this study provides a comprehensive investigation of biochar (derived from mangrove Kandelia obovata -sediment system under three conditions: I) co-addition of biochar and sediment; II) biochar and subsequently sediment addition (after biochar adsorption reached equilibrium); III) sediment and subsequently biochar addition (after sediment adsorption reached equilibrium). It was observed that the adsorption capability under model I and III was much smaller than that under model II (p < 0.05). Regardless of time, K. obovate - biochar significantly (p < 0.05) increase the sorption of phenanthrene in sediment -water system. The results provide valuable studies for further in-situ remediation of phenanthrene and engineering applications.

Electrochemical aptamer-based determination of protein tyrosine kinase-7 using toehold-mediated strand displacement amplification on gold nanoparticles and graphene oxide.

An electrochemical method is described for ultrasensitive determination of protein tyrosine kinase-7 (PTK7). It is based on (a) the use of positively charged gold nanoparticles (AuNPs) and negatively charged graphene oxide (GO), and (b) of toehold-mediated strand displacement amplification. A hairpin probe 2 (HP2) containing the sgc8 aptamer was used to modify a glassy carbon electrode (GCE). Its hairpin structure is opened in the presence of PTK7 to form the PTK7-HP2 complex. The exposed part of HP2 partly hybridizes with hairpin probe 1 (HP1) that was immobilizing on the AuNPs and GO modified GCE. On addition of the hairpin probe 3 that was labeled with the redox probe Methylene Blue (MB-HP3), toehold-mediated strand displacement occurs due to complementary hybridization of HP1 with MB-HP3. This causes the release of PTK7-HP2 into the solution and makes it available for the next reaction. Under optimal conditions, PTK7 can be quantified by voltammetry (typically performed at -0.18 V) with a detection limit of 1.8 fM. The assay possesses high selectivity for PTK7 due to the employment of the aptamer. It was successfully applied to the determination of PTK7 in the debris of malignant melanoma A375 cells. Graphical abstract Schematic representation of the enzyme-free electrochemical sensor for ultrasensitive determination of protein tyrosine kinase-7 (PTK7) based on the toehold-mediated strand displacement reaction amplification on gold nanoparticles and graphene oxide.

Simultaneous decolorization of sulfonated azo dyes and reduction of hexavalent chromium under high salt condition by a newly isolated salt-tolerant strain Bacillus circulans BWL1061.

The co-existence of dyes, Cr(VI) and high concentration of salt in dyeing wastewater causes serious and complex environmental problems. In this study, a salt-tolerant strain Bacillus circulans BWL1061 was reported to simultaneously remove 50mg/L methyl orange and 50mg/L Cr(VI) under the anaerobic condition with 60g/L NaCl. During the decolorization process, the Cr(VI) reduction occurred preferentially over the dye decolorization due to the dominate utilization of electron by Cr(VI). The analysis of enzyme activities suggested that azoreductase, NADH-DCIP reductase, and laccase were associated with decolorization of methyl orange. A possible degradation pathway was proposed based on the metabolites analysis. The decolorization of methyl orange is involved in the symmetric cleavage of azo bond, which formed N,N-dimethyl p-phenylenediamine and 4-amino sulfonic acid, or the asymmetric cleavage of azo bond, which formed 4-(dimethylamino) phenol and 4-diazenylbenzene sulfonic acid. Phytotoxicity assays showed that strain BWL1061 could decrease the toxicity of methyl orange to Triticum aestivum, Pogostemon cablin and Isatis indigotica Fort during the decolorization process. In this study, Bacillus circulans is reported for the first time that could simultaneously remove azo dyes and Cr (VI) under high salt condition.

The weighted multi-scale connections networks for macrodispersivity estimation.

Macrodispersivity is critical for predicting solute behaviors with dispersive transport models. Conventional methods of estimating macrodispersivity usually need to solve flow equations and are time-consuming. Convolutional neural networks (CNN) have recently been proven capable of efficiently mapping the hydraulic conductivity field and macrodispersivity. However, the mapping accuracy still needs further improvement. In this paper, we present a new network shortcut connection style called weighted multi-scale connections (WMC) for convolutional neural networks to improve mapping accuracy. We provide empirical evidence showing that the WMC can improve the performance of CNN in macrodispersivity estimation by implementing the WMC in CNNs (CNN without short-cut connections, ResNet, and DenseNet), and evaluating them on datasets of macrodispersivity estimation. For the CNN without short-cut connections, the WMC can improve the estimating R2 by at least 3% on three datasets of conductivity fields. For ResNet18, the WMC improved the estimated R2 by an average of 2.5% on all three datasets. For ResNet34, the WMC improved the estimated R2 by an average of 5.6%. For ResNet50, the WMC improved the estimated R2 by an average of 16%. For ResNet101, the WMC improved the estimating R2 by an average of 30%. For DenseNets, the improved estimated R2 ranges from 0.5% to 5%. The WMC can strengthen feature propagation of different sizes and alleviate the vanishing-gradient issue. Moreover, it can be implemented to any CNN with down-sampling layers or blocks.

Efficacy and underlying mechanisms of three-circle post standing qigong on insomnia in college students: a four-arm, double-blind, randomized controlled trial protocol.

BACKGROUND: Insomnia is common in college students, but its impact on health and wellbeing is often neglected. Enhancing sleep quality through targeted interventions could improve overall health and reduce the risk of consequent co-morbidities and mental health problems. Qigong exercises have been shown to significantly improve sleep quality and relieve insomnia. Three-circle Post Standing (TCPS) can help integrate body, breath, and mind, a fundamental principle of Qigong that promotes holistic wellbeing. In this clinical trial, we aim to (1) evaluate the feasibility, safety, and therapeutic efficacy of administering TCPS to improve sleep quality and quality of life in college students with insomnia; (2) explore the neurophysiological mechanisms underlying the mind adjustments mediated by TCPS in insomnia; (3) investigate body and breath pathophysiology mediated by TCPS in insomnia; and (4) assess the long-term efficacy of TCPS in terms of sleep quality and quality of life. METHODS: This will be a prospective, parallel, four-arm, double-blind randomized controlled trial to investigate the effects and underlying mechanisms of TCPS on college students with insomnia. One hundred college students meeting diagnostic criteria for insomnia will be randomly assigned to receive either 14 weeks of standardized TCPS training (two weeks of centralized training followed by 12 weeks of supervised training) or sham-control Post Standing training. Efficacy outcomes including sleep quality, quality of life, neurophysiological assessments, plantar pressure, biomechanical balance, and physical measures will be collected at baseline, eight weeks (mid-point of supervised training), and 14 weeks (end of supervised training). Sleep quality and quality of life will also be evaluated during the four- and eight-week follow-up. DISCUSSION: This trial will be an important milestone in the development of new therapeutic approaches for insomnia and should be easily implementable by college students with insomnia. The neuro- and pathophysiological assessments will provide new insights into the mechanisms underlying TCPS. CLINICAL TRIAL REGISTRATION: This trial has been registered in the China Clinical Trials Registry (registration number: ChiCTR2400080763).

Dual Phytochemical/Activity-Guided Optimal Preparation and Bioactive Material Basis of Orthosiphon Stamineus Benth. (Shen Tea) against Nonalcoholic Fatty Liver Disease.

Orthosiphon stamineus Benth. (OSB) is a popular plant used for making "Shen tea" or "Java tea". It has been demonstrated with antioxidant, anti-inflammatory, and hepatoprotective activities. However, its potential beneficial effects and bioactive material basis for nonalcoholic fatty liver disease (NAFLD) has not been convincingly studied. In the present work, we conducted dual phytochemical/activity-guided extraction optimization and component fractionation of OSB, and evaluated its beneficial effects on NAFLD. Flavonoids and polyphenols (caffeic acid/protocatechuic acid derivatives) were determined as the dominant phytochemicals in OSB. The extraction process for these phytochemicals was optimized by using response surface methodology. Noticeably, flavonoids showed a stronger correlation with the antioxidant activities of OSB than polyphenols. Likewise, the flavonoid-rich fraction of OSB exerted antioxidant activities stronger than those of other fractions. As expected, in vitro and in vivo studies demonstrated that the flavonoid-rich fraction effectively attenuated weight increase, improved lipid metabolism, alleviated hepatic steatosis, and reversed hepatic inflammation. Importantly, this fraction showed equivalent beneficial effects to the total extract of OSB, suggesting that flavonoids were the main bioactive constituents of OSB. The action mechanism was indicated as direct antioxidant effect through chemical interaction with free radicals and indirect mitochondria-mediated antioxidant defense. Our research offers bioactive substances for further exploitation and expands the potential application of OSB.

Effects of local transverse dispersion on macro-scale coefficients of oxygen-limited biodegradation in a stratified formation.

The correct characterization of macro-scale contaminant transport and transformation rates is an important issue for modeling reactive transport in heterogeneous aquifers. While previous studies have investigated field-scale heterogeneity of transport and biochemical properties, the effects of local transverse dispersion on macro-scale transport and transformation rates have not been well understood. In this paper, the process of oxygen-limited biodegradation in a stratified aquifer is analysed by spectral perturbation approach, and longitudinal macrodispersivity, effective biodegradation rate, effective retardation factor and effective velocity are derived for the coupled transport equations of a system consisting of a contaminant and an oxidizing agent (oxygen). The effects of local transverse dispersion on these macro-scale coefficients are studied. It is shown that local transverse dispersion can smooth the heterogeneity in biodegradation and sorption processes and enlarge effective biodegradation rate and retardation factor. The local transverse dispersion can also limit the effects of heterogeneity in biodegradation process on longitudinal macrodispersivities and effective velocities for the contaminant and dissolved oxygen. But the effects of heterogeneity in sorption process on the contaminant macrodispersivity is likely to be magnified by local transverse dispersion.

Characterisation and functional analysis of canine TLR5.

In this study, we characterised the single exon TLR5 gene of the Chinese rural dog. Sequence analysis revealed a 2577 nucleotide-long open reading frame of canine TLR5, encoding an 858 amino acid-long protein. The putative amino acid sequence of canine TLR5 consisted of a signal peptide sequence, 15 LRR domains, a LRR C-terminal domain, a transmembrane domain and an intracellular Toll-IL-1 receptor domain. The amino acid sequence of the canine TLR5 protein shared 95.4% identity with vulpine, 72.2% with feline and 64.7% with human TLR5. Plasmids expressing canine TLR5 and NF-κB-luciferase were constructed and transfected into HEK293T cells. Expression was confirmed by indirect immunofluorescence assay. These HEK293T cells transfected with the canine TLR5- and NF-κB-luciferase plasmids significantly responded to flagellin from Salmonella enteritidis serovar Typhimurium, indicating that it is a functional TLR5 homolog. In response to stimulation with Salmonella enteritidis, the level of TLR5 mRNA significantly increased over the control in PBMCs at 4 h. The levels of IL-8, IL-6 and IL-1ß also increased after exposure. The highest levels of TLR5, IL-8 and IL-1ß expression were detected at 8, 4 and 12 h after stimulation, respectively. These results imply that the expression of canine TLR5 may participate in the immune response against bacterial pathogens.

Identification of candidate miRNAs related in storage root development of sweet potato by high throughput sequencing.

Sweet potato (Ipomoea batatas L.) is a food consumed worldwide, an industrial raw material and new energy crop. The storage root is the most economical part of the crop. However, the mechanism of storage root initiation and development is still unclear. In this study, conserved and novel miRNAs during storage root development were identified by high-throughput sequencing technology by constructing small RNA libraries from sweet potato fibrous roots (F) and storage roots at four different developmental stages (storage roots with different diameters: 1 cm, D1; 3 cm, D3; 5 cm, D5 and 10 cm, D10). A total of 61 known miRNAs and 471 novel miRNAs were identified. In addition, 145 differentially expressed miRNAs were identified in the F library compared with the four storage root libraries, with 30 known miRNAs and 115 novel miRNAs. Moreover, the targets of the differentially expressed miRNAs were predicted and their network was further investigated by GO analysis using our previous transcriptome data. The GO analysis revealed that antioxidant activity and binding process were the most enriched terms of the target genes. The secondary structure and expression of six candidate miRNAs including three conserved miRNAs and three novel miRNAs were investigated and their predicted targets were validated by qRT-PCR. The results showed that the expression levels of the miRNAs were all consistent with the sequencing data. Most of the miRNAs and their corresponding targets had obvious negative correlations. This study contributed to elucidating the potential miRNA mediated regulatory mechanism of storage root development in sweet potato. The specific differentially expressed miRNAs in sweet potato storage roots can be used to breed high-yield sweet potatoes and other tuberous root crops.

Prevalence and Drug Resistance of Salmonella in Dogs and Cats in Xuzhou, China.

INTRODUCTION: Salmonellosis is a zoonotic disease, and Salmonella spp. can sometimes be found in dogs and cats, posing a risk to human health. In this study, the prevalence and antimicrobial susceptibility of faecal Salmonella were investigated in pet dogs and cats in Xuzhou, Jiangsu Province, China. MATERIAL AND METHODS: Faecal samples from 243 dogs and 113 cats, at seven pet clinics, were tested between March 2018 and May 2019. Each Salmonella isolate was characterised using serotyping and antimicrobial susceptibility tests. RESULTS: The prevalence of Salmonella was 9.47% in dogs and 1.77% in cats. Among the 25 isolates, eight serotypes of Salmonella enterica subsp. enterica were detected, S. Kentucky (n = 11), S. Indiana (n = 5), and S. Typhimurium (n = 4) predominating. S. Derby, S. Toucra, S. Sandiego, S. Newport, and S. Saintpaul all occurred singly. The 23 Salmonella strains found in dogs were from seven different serovars, while the two strains in cats were from two. The highest resistance rates were found for tetracycline (92%), azithromycin (88%), cefazolin (84%), nalidixic acid (80%), ampicillin (80%), ceftriaxone (80%), and streptomycin (76%). Resistance to three or more antimicrobial agents was detected in 24 (96%) isolates. Most of the S. Kentucky and S. Indiana isolates were multi-drug resistant to more than 11 agents. CONCLUSION: The carriage rate was far higher in dogs than in cats from Xuzhou. Some isolated strains were highly resistant to antimicrobials used to treat infections in humans and pets, which may raise the risk of humans being infected with multi-drug resistant Salmonella via close contact with pets.

Genome-wide identification and expression analysis of glycine-rich RNA-binding protein family in sweet potato wild relative Ipomoea trifida.

Glycine-rich RNA-binding proteins (GRPs) contain RNA recognition motif (RRM) and glycine-rich domains at the N- or C-terminus, respectively, and they participate in varied physiological and biochemical processes, as well as environmental stresses. Sweet potato from the genus Ipomoea is one of the most important crops. However, the role of the GRP gene family in Ipomoea plant species has not been reported yet. At the same time, the genome of sweet potato remains to be elucidated, but the genome of I. trifida which is most probably the progenitor of the sweet potato was released recently. In this regard, we carried out genome-wide analysis of GRP family members in I. trifida. Here, we identified nine GRP genes in I. trifida and investigated their motif distribution, promoters and gene structure. Subsequently, we performed phylogenetic analysis with the GRP genes from I. trifida, Arabidopsis thaliana, Zea mays L. and Oryza sativa to investigate their phylogenetic relationship. Moreover, we studied the expression patterns of ItGRPs in the roots, stems, young and mature leaves and flowers and found that ItGRP genes were tissue-specific. Meanwhile, the expression profiles under four abiotic stress conditions, including heat, cold, salt and drought stress treatments, revealed that some genes were markedly up-regulated or down-regulated. Taken together, our findings will provide reference to studies on the function of GRP genes in the development and stress response of I. trifida.

Comprehensive RNA-Seq Data Analysis Identifies Key mRNAs and lncRNAs in Atrial Fibrillation.

Long non-coding RNAs (lncRNAs) are an emerging class of RNA species that may play a critical regulatory role in gene expression. However, the association between lncRNAs and atrial fibrillation (AF) is still not fully understood. In this study, we used RNA sequencing data to identify and quantify the both protein coding genes (PCGs) and lncRNAs. The high enrichment of these up-regulated genes in biological functions concerning response to virus and inflammatory response suggested that chronic viral infection may lead to activated inflammatory pathways, thereby alter the electrophysiology, structure, and autonomic remodeling of the atria. In contrast, the downregulated GO terms were related to the response to saccharides. To identify key lncRNAs involved in AF, we predicted lncRNAs regulating expression of the adjacent PCGs, and characterized biological function of the dysregulated lncRNAs. We found that two lncRNAs, ETF1P2, and AP001053.11, could interact with protein-coding genes (PCGs), which were implicated in AF. In conclusion, we identified key PCGs and lncRNAs, which may be implicated in AF, which not only improves our understanding of the roles of lncRNAs in AF, but also provides potentially functional lncRNAs for AF researchers.

The effect of Three-Circle Post Standing (Zhanzhuang) Qigong on the physical and psychological well-being of college students: Study protocol for a randomized controlled trial.

BACKGROUND: The physical and mental health of college students tends to continuously decline around the world due to lifestyle or behavior habits changes, and pervasive presence of the Internet. Thus it is urgent to improve their health in college life. As a traditional Qigong form is practiced mainly in a standing posture. Three-Circle Post Standing Qigong (TCPSQ) is suitable for regular practicing and has beneficial effects on improving the physiological function and psychological emotion by adjusting body, breathing, and mind. The aim of the 3 adjustments is to achieve a state of harmonious unity-integrating these adjustments into "one." In this study protocol article, we will systematically explore the effectiveness and safety, feasibility of TCPSQ on physical and psychological outcomes of the college students and deeply understand the state of harmonious unity-integrating adjustments of body, breath, and mind into "one." METHOD/DESIGN: We will conduct a randomized, assessor, and statistician blinded, parallel-controlled trial comparing the beneficial effect of TCPSQ in college students. A total of 80 eligible college students from the Beijing University of Chinese Medicine (BUCM) will be recruited and randomly allocated into TCPSQ training or unaltered lifestyle control group according 1:1 allocation ratio with allocation concealment. TCPSQ intervention will last 10 weeks. The study period is 18 weeks including a 10-week supervised intervention and a 8-week follow-up. The relevant physical and psychological outcomes, adverse events, and safety will be evaluated at baseline, 6 weeks (at the mid-point of intervention), 11 weeks (at the end of intervention), and 19 weeks (after the 8-week follow-up period) by blinded independent outcome assessors. DISCUSSION: This is the first randomized controlled trial protocol from the perspective of Qigong connotation to systematically investigate the effect of TCPSQ for the physical and mental health of a college student population. If the results in our study prove a significant intervention effect, this would provide preliminary higher-quality evidence and establish an optimal guidance for the application of TCPSQ exercise program among a college student population. ETHICS AND DISSEMINATION: The study was approved by the ethics committee of the Beijing University of Chinese Medicine (approval number: 2018BZHYLL0109). A SPIRIT checklist is available for this protocol. The trial was registered in Chinese Clinical Trial Registry (WHO ICTRP member). Registration number: ChiCTR-BON-17010840.

Synthesis and characterization of nontoxic chitosan-coated Fe3O4 particles for patulin adsorption in a juice-pH simulation aqueous.

Chitosan-coated Fe3O4 particles were prepared as a magnetic adsorbent by reverse oil-in-water micro-emulsion system using Triton X-100 as the emulsifier. Coating chitosan onto the magnetic particles was confirmed by transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectra and magnetic measurements. Chitosan-coated Fe3O4 adsorbent was shown to be effective for patulin adsorption with a maximum adsorption capacity of 6.67mg/g within 5h by adding 300µg adsorbents into 10mL 200µg/L patulin aqueous. In addition, the recovery rate of chitosan-coated Fe3O4 adsorbent reached to 99.95% within 60min, showed its excellent recoverable performance. Moreover, in vitro cytotoxicity and acute toxicity evaluation were also conducted, the results suggested that the chitosan-coated Fe3O4 adsorbent was non-cytotoxic, and had no toxic response or histopathological changes on mice. The results of this study demonstrated that chitosan-coated Fe3O4 particles are promising adsorbents for patulin removal in fruit juice industry.