Identification of WRKY gene family members in amaranth based on a transcriptome database and functional analysis of AtrWRKY42-2 in betalain metabolism

IntroductionWRKY TFs (WRKY transcription factors) contribute to the synthesis of secondary metabolites in plants. Betalains are natural pigments that do not coexist with anthocyanins within the same plant. Amaranthus tricolor (‘Suxian No.1’) is an important leaf vegetable rich in betalains. However, the WRKY family members in amaranth and their roles in betalain synthesis and metabolism are still unclear.MethodsTo elucidate the molecular characteristics of the amaranth WRKY gene family and its role in betalain synthesis, WRKY gene family members were screened and identified using amaranth transcriptome data, and their physicochemical properties, conserved domains, phylogenetic relationships, and conserved motifs were analyzed using bioinformatics methods.ResultsIn total, 72 WRKY family members were identified from the amaranth transcriptome. Three WRKY genes involved in betalain synthesis were screened in the phylogenetic analysis of WRKY TFs. RT-qPCR showed that the expression levels of these three genes in red amaranth ‘Suxian No.1’ were higher than those in green amaranth ‘Suxian No.2’ and also showed that the expression level of AtrWRKY42 gene short-spliced transcript AtrWRKY42-2 in Amaranth ‘Suxian No.1’ was higher than that of the complete sequence AtrWRKY42-1, so the short-spliced transcript AtrWRKY42-2 was mainly expressed in ‘Suxian No.2’ amaranth. Moreover, the total expression levels of AtrWRKY42-1 and AtrWRKY42-2 were down-regulated after GA3 treatment, so AtrWRKY42-2 was identified as a candidate gene. Therefore, the short splice variant AtrWRKY42-2 cDNA sequence, gDNA sequence, and promoter sequence of AtrWRKY42 were cloned, and the PRI 101-AN-AtrWRKY42-2-EGFP vector was constructed to evaluate subcellular localization, revealing that AtrWRKY42-2 is located in the nucleus. The overexpression vector pRI 101-AN-AtrWRKY42-2-EGFP and VIGS (virus-induced gene silencing) vector pTRV2-AtrWRKY42-2 were transferred into leaves of ‘Suxian No.1’ by an Agrobacterium-mediated method. The results showed that AtrWRKY42-2 overexpression could promote the expression of AtrCYP76AD1 and increase betalain synthesis. A yeast one-hybrid assay demonstrated that AtrWRKY42-2 could bind to the AtrCYP76AD1 promoter to regulate betalain synthesis.DiscussionThis study lays a foundation for further exploring the function of AtrWRKY42-2 in betalain metabolism

Interfacial rheology of coextruded elastomeric and amorphous glass thermoplastic polyurethanes

Abstract In this work, we report on the sensitivity of rheometrical techniques to the nature and size of the interface/interphase in coextruded thermoplastic urethanes (TPUs). In particular, the interphases developed during coextrusion of an amorphous glass (hard) TPU (Isoplast ® ETPU 301) with one of two elastomeric (soft) TPUs (Estane ® TPU 58277 and Estane ® TPU X1175) were studied. Differences in the thickness and nature of the interphase of the two coextruded bilayer films were observed by atomic force microscopy. In one case, the interphase is thicker and rough, and in the other case, it is thinner and flat. Rheology was used in order to probe the type and characteristics of the interphases, with coextruded films having been tested in steady shear, small-amplitude oscillatory shear (SAOS), uniaxial extension, and stress relaxation after a step strain in shear. The results were compared with theoretical predictions assuming zero-thickness interfaces and no interfacial slip. For SAOS and stress relaxation experiments, expressions were deduced in order to enable such a prediction to be made. Of all four rheometrical tests, only stress relaxation after a step shear did not follow the theoretical predictions and, thus, was sensitive enough to detect the presence of the interphase

Knockdown of Midgut Genes by dsRNA-Transgenic Plant-Mediated RNA Interference in the Hemipteran Insect Nilaparvata lugens

BACKGROUND: RNA interference (RNAi) is a powerful technique for functional genomics research in insects. Transgenic plants producing double-stranded RNA (dsRNA) directed against insect genes have been reported for lepidopteran and coleopteran insects, showing potential for field-level control of insect pests, but this has not been reported for other insect orders. METHODOLOGY/PRINCIPAL FINDINGS: The Hemipteran insect brown planthopper (Nilaparvata lugens Stål) is a typical phloem sap feeder specific to rice (Oryza sativa L.). To analyze the potential of exploiting RNAi-mediated effects in this insect, we identified genes (Nlsid-1 and Nlaub) encoding proteins that might be involved in the RNAi pathway in N. lugens. Both genes are expressed ubiquitously in nymphs and adult insects. Three genes (the hexose transporter gene NlHT1, the carboxypeptidase gene Nlcar and the trypsin-like serine protease gene Nltry) that are highly expressed in the N. lugens midgut were isolated and used to develop dsRNA constructs for transforming rice. RNA blot analysis showed that the dsRNAs were transcribed and some of them were processed to siRNAs in the transgenic lines. When nymphs were fed on rice plants expressing dsRNA, levels of transcripts of the targeted genes in the midgut were reduced; however, lethal phenotypic effects after dsRNA feeding were not observed. CONCLUSIONS: Our study shows that genes for the RNAi pathway (Nlsid-1 and Nlaub) are present in N. lugens. When insects were fed on rice plant materials expressing dsRNAs, RNA interference was triggered and the target genes transcript levels were suppressed. The gene knockdown technique described here may prove to be a valuable tool for further investigations in N. lugens. The results demonstrate the potential of dsRNA-mediated RNAi for field-level control of planthoppers, but appropriate target genes must be selected when designing the dsRNA-transgenic plants

Effects of bisphosphonates in preventing periprosthetic bone loss following total hip arthroplasty: a systematic review and meta-analysis

Abstract Background Periprosthetic bone loss following total hip arthroplasty (THA) was a well-known phenomenon. This systematic review was to assess the effectiveness of bisphosphonates (BPs) for decreasing periprosthetic bone resorption. Methods The MEDLINE, EMBASE, and Cochrane Library databases were searched up to March 2018. Randomized controlled trials compared the effects between administrating BPs and placebo or no medication were eligible; the target participants were patients who underwent THA. Mean differences (MD) and 95% confidence interval (95% CI) were calculated by using the random-effects models. Statistical analyses were performed by RevMan 5.3 software. Results Fourteen trials involving 620 patients underwent THA were retrieved. BPs significantly prevented the loss of periprosthetic bone mineral density at 1 year (MD, 0.06 [95% CI, 0.03 to 0.08], p < 0.001), between 2 and 4 years (MD, 0.04 [95% CI, 0.01 to 0.07], p = 0.02), and more than 5 years after THA (MD, 0.08 [95% CI, 0.06 to 0.11], p < 0.001). Both serum bone alkaline phosphatase (MD, − 7.28 [95% CI, − 9.81 to − 4.75], p < 0.001) and urinary N-telopeptide of type I collagen (MD, − 24.37 [95% CI, − 36.37 to − 12.37], p < 0.001) in BP group were significantly lower. Subgroup analyses showed that the third-generation BPs were more effective in decreasing periprosthetic bone loss than the first and second generation within 1 year after THA (p = 0.001). Conclusion BPs were beneficial to decreasing periprosthetic bone loss. The third-generation BPs showed significantly efficacy for patients in short-term observation

Industrial cluster, government agency and entrepreneurial development: a case study of Wenzhou City, Zhejiang Province

Purpose - The purpose of the paper is to explore why the entrepreneurial activities are agglomerative in a cluster and to investigate the government agencies' functions in the industry cluster that can construct favourable environment for entrepreneurial development.Design/methodology/approach - The case study method is chosen to collect in-depth data to investigate the research domain, and the detailed case of the industrial cluster in Wenzhou, China is selected. The empirical data for the analysis are derived from in-depth interviews with entrepreneurs in private and public organizations. Statistical data and historical documents are also collected to increase the understanding of the regional conditions, as well as for comparison with and triangulation of the research theme.Findings - It was found that in Wenzhou, four major groups of government agencies which perform the functions of investment, research & innovation, industrial information and supporting service, were generated along with the development of the industrial clusters; and it was found that initial capital, technology support and human capital are the critical resources those institutes had tried to provide to facilitate the entrepreneurial activities in the local clusters.Originality/value - The paper enriches our understanding about how entrepreneurship is promoted and cultivated in industrial clusters under the social-economic environment in China and sheds light on the Chinese entrepreneurial process in the local industrial clusters

A Novel Hypoxia Related Marker in Blood Link to Aid Diagnosis and Therapy in Osteoarthritis

Osteoarthritis (OA) is a common chronic degenerative arthritis. Its treatment options are very limited. At present, hypoxia is a prominent factor in OA. This study aimed to re-explore the mechanism between hypoxia and OA, which provides new insights into the diagnosis and therapy of OA. We acquired the OA-related expression profiles of GSE48556, GSE55235, and GSE55457 for our analysis. Using gene set variation analysis (GSVA), we found significant differences in hypoxia. These differences result from multiple pathways, such as the p53 signaling pathway, cell senescence, the NF-kappa B signaling pathway, Ubiquitin-mediated proteolysis, and apoptosis. Meanwhile, the single-sample gene set enrichment analysis (ssGSEA) showed that hypoxia was significantly associated with the level of immune cell infiltration in the immune microenvironment. Thus, we believe that hypoxia is useful for the diagnosis and treatment of OA. We successfully constructed a novel hypoxia-related index (HRI) based on seven hypoxia-related genes (ADM, CDKN3, ENO1, NDRG1, PGAM1, SLC2A1, VEGFA) by least absolute shrinkage and binary logistic regression of the generalized linear regression. HRI showed potential for improving OA diagnosis through receiver operation characteristic (ROC) analysis (AUC training cohort = 0.919, AUC testing cohort = 0.985). Moreover, we found that celastrol, droxinostat, torin-2, and narciclasine may be potential therapeutic compounds for OA based on the Connectivity Map (CMap). In conclusion, hypoxia is involved in the development and progression of OA. HRI can improve diagnosis and show great potential in clinical application. Celastrol, droxinostat, torin-2, and narciclasine may be potential compounds for the treatment of OA patients

Arsenic (V) adsorption on Fe3O4 nanoparticle-coated boron nitride nanotubes

Multiwalled boron nitride nanotubes (BNNTs) functionalized with Fe(3)O(4) nanoparticles (NPs) were used for arsenic removal from water solutions. Sonication followed by a heating process was developed to in situ functionalize Fe(3)O(4) NPs onto a tube surface. A batch of adsorption experiments conducted at neutral pH (6.9) and room temperature (25 degrees C) and using the developed nanocomposites revealed effective arsenic (V) removal. The Langmuir, Freundlich, and Dubinin-Radushkevich adsorption isotherms were measured for a range of As(V) initial concentrations from 1 to 40 mg/L under the same conditions. The equilibrium data well fitted all isotherms, indicating that the mechanism for As(V) adsorption was a combination of chemical complexation and physical electrostatic attraction with a slight preference for chemisorption. The magnetite NPs functionalized on BNNTs led to a simple and rapid separation of magnetic metal-loaded adsorbents from the treated water under an external magnetic field

Sensitive and rapid detection of tetrodotoxin based on gold nanoflower-and latex microsphere-labeled monoclonal antibodies

Tetrodotoxin (TTX) could result in serious diseases due to its extremely high neurotoxicity. Thus, it is of great importance to measure TTX for food safety. In this study, an anti-TTX monoclonal antibody with good specificity and high affinity was used to develop the immunochromatographic test strips (ICTS). Gold nanoflower (AuNF) with multiple branches and latex microsphere (LM) with large particle size as signal reporters were employed for improving the sensitivity of test strips. Both AuNF and LM probes are stable, and the developed ICTS were specific to TTX, demonstrating no cross-reactivity with other marine toxins. The linear range of AuNF- and LM-based strips for TTX was 9.49–330.98 ng/mL and 5.40–443.19 ng/mL, respectively. The limit of detection (LOD) of AuNF- and LM-based strips was determined to be 9.49 ng/mL and 5.40 ng/mL, respectively. In summary, the developed ICTS based on AuNF and LM signal probes displayed enhancement of sensitivity and provided rapid and specific detection of TTX