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Postharvest deterioration is among the major challenges for the fruit industry. Regulation of the fruit softening rate is an effective strategy for extending shelf-life and reducing the economic losses due postharvest deterioration. The tomato myoinositol monophosphatase 3 gene SlIMP3, which showed highest expression level in fruit, was expressed and purified. SlIMP3 demonstrated high affinity with the L-Gal 1-P and D-Ins 3-P, and acted as a bifunctional enzyme in the biosynthesis of AsA and myoinositol. Overexpression of SlIMP3 not only improved AsA and myoinositol content, but also increased cell wall thickness, improved fruit firmness, delayed fruit softening, decreased water loss, and extended shelf-life. Overexpression of SlIMP3 also increased uronic acid, rhamnose, xylose, mannose, and galactose content in cell wall of fruit. Treating fruit with myoinositol obtained similar fruit phenotypes of SlIMP3-overexpressed fruit, with increased cell wall thickness and delayed fruit softening. Meanwhile, overexpression of SlIMP3 conferred tomato fruit tolerance to Botrytis cinerea. The function of SlIMP3 in cell wall biogenesis and fruit softening were also verified using another tomato species, Ailsa Craig (AC). Overexpression of SlDHAR in fruit increased AsA content, but did not affect the cell wall thickness or fruit firmness and softening. The results support a critical role for SlIMP3 in AsA biosynthesis and cell wall biogenesis, and provide a new method of delaying tomato fruit softening, and insight into the link between AsA and cell wall metabolism.

The purpose of this study was to investigate the effects of a procedure involving systematic withdrawal of stimulus prompts and tangible reinforcers on the acquisition, maintenance, and generalization of eye contact and gaze following for two children with ASD in China. Two boys with ASD (5–6 years of age) participated. A concurrent multiple probe design across behaviors and participants was used. Results indicate that the procedure effectively established eye contact and gaze following for both children. Generalization to new instructors occurred in the free play setting, and the acquired behaviors were maintained for 1 month following training. Eye contact was maintained with social consequences for one child; the other child required tokens along with social consequences to maintain eye contact. Social consequences were sufficient to maintain gaze following for both children.

The wide application of optical spectroscopy makes miniaturized spectrometers with fundamental importance. The scalability, high-performance, low-cost, and small footprint are still contradicting each other and limiting the applicability of miniaturized spectrometer for practical application. Here we propose a compact spectrometer that satisfies the four advantages. The device uses a fiber taper tip to generate complex leaky mode patterns within 1 mm length. The unique correspondence between the pattern and wavelength operates effectively for hundreds of nanometers spectral range while providing a spectral resolution around ~ 1 pm. The integration of multiple taper tips enables hyperspectral imaging applications. The working range of our device can be further extended using different materials and detectors while keeping the similar architecture.

Vanadium dioxide (VO2) glass is a promising thermochromic smart energy-saving window. Many studies have been carried out for improving its optical performance, but for practical applications, the w...

Summary Unicellular and multicellular tomato trichomes function as mechanical and chemical barriers against herbivores. Auxin treatment increased the formation of II, V and VI type trichomes in tomato leaves. The auxin response factor gene SlARF4, which was highly expressed in II, V and VI type trichomes, positively regulated the auxin‐induced formation of II, V and VI type trichomes in the tomato leaves. SlARF4 overexpression plants with high densities of these trichomes exhibited tolerance to spider mites. Two R2R3 MYB genes, SlTHM1 and SlMYB52, were directly targeted and inhibited by SlARF4. SlTHM1 was specifically expressed in II and VI type trichomes and negatively regulated the auxin‐induced formation of II and VI type trichomes in the tomato leaves. SlTHM1 down‐regulation plants with high densities of II and VI type trichomes also showed tolerance to spider mites. SlMYB52 was specifically expressed in V type trichomes and negatively regulated the auxin‐induced formation of V type trichome in the tomato leaves. The regulation of SlARF4 on the formation of II, V and VI type trichomes depended on SlTHM1 and SlMYB52, which directly targeted cyclin gene SlCycB2 and increased its expression. In conclusion, our data indicates that the R2R3 MYB‐dependent auxin signalling pathway regulates the formation of II, V and VI type trichomes in tomato leaves. Our study provides an effective method for improving the tolerance of tomato to spider mites.

Burkholderia pyrrocinia JK-SH007 can effectively control poplar canker caused by pathogenic fungi. Its antifungal mechanism remains to be explored. Here, we characterized the functional role of CysB in B. pyrrocinia JK-SH007. This protein was shown to be responsible for the synthesis of cysteine and the siderophore ornibactin, as well as the antifungal activity of B. pyrrocinia JK-SH007. We found that deletion of the cysB gene reduced the antifungal activity and production of the siderophore ornibactin in B. pyrrocinia JK-SH007. However, supplementation with cysteine largely restored these two abilities in the mutant. Further global transcriptome analysis demonstrated that the amino acid metabolic pathway was significantly affected and that some sRNAs were significantly upregulated and targeted the iron–sulfur metabolic pathway by TargetRNA2 prediction. Therefore, we suggest that, in B. pyrrocinia JK-SH007, CysB can regulate the expression of genes related to Fe–S clusters in the iron–sulfur metabolic pathway to affect the antifungal activity of B. pyrrocinia JK-SH007. These findings provide new insights into the various biological functions regulated by CysB in B. pyrrocinia JK-SH007 and the relationship between iron–sulfur metabolic pathways and fungal inhibitory substances. Additionally, they lay the foundation for further investigation of the main antagonistic substances of B. pyrrocinia JK-SH007.

It is critical for tourism managers and marketers to understand how to support the sustainable recovery of the industry by involving residents in tourism marketing. The purpose of this study is to investigate whether the COVID-19 pandemic, as an environmental stimulus, could enhance the salience of residents’ collective identity, thereby increasing their engagement with place branding. A conceptual framework was developed to explore the role of relationship-inducing factors and non-relationship-inducing factors in activating residents’ collective identity based on the social identity salience theory of relationship marketing success. The research model was tested using data from 603 questionnaires collected from Sanya, China, during the period of a sudden COVID-19-related crisis. The findings from the statistical analyses showed significant relationships among the research variables, and the moderating role of perceived social connectedness was supported. Our findings have implications regarding how to engage residents in the tourism recovery process during and post the COVID-19 pandemic.

Autonomic dysfunction is common in transthyretin familial amyloid polyneuropathy (TTR-FAP). Because ultrasonography is a powerful tool to study peripheral neuropathy, vagus nerve (VN) ultrasonography was used in our study to investigate the possible changes of the dimension of VN in TTR-FAP.Eighteen patients with TTR-FAP and 17 age- and gender-matched individuals without any neuropathies were enrolled in a pilot study. The cross-sectional areas (CSAs) were measured bilaterally on transverse scans of vagus, median, and ulnar nerves. Clinical data were collected to explore the correlations with CSAs of VN.The median CSAs of VN in TTR-FAP were 3.5 (2.0-6.0) mmVN enlargement is prevalent among TTR-FAP patients. VN ultrasonography may be an important clinical tool for assessing the severity of autonomic dysfunction in TTR-FAP.

Incorporating passive radiative cooling structures into personal thermal management technologies could effectively defend humans against intensifying global climate change. We show that large-scale woven metafabrics can provide high emissivity (94.5%) in the atmospheric window and high reflectivity (92.4%) in the solar spectrum because of the hierarchical-morphology design of the randomly dispersed scatterers throughout the metafabric. Through scalable industrial textile manufacturing routes, our metafabrics exhibit desirable mechanical strength, waterproofness, and breathability for commercial clothing while maintaining efficient radiative cooling ability. Practical application tests demonstrated that a human body covered by our metafabric could be cooled ~4.8°C lower than one covered by commercial cotton fabric. The cost-effectiveness and high performance of our metafabrics present substantial advantages for intelligent garments, smart textiles, and passive radiative cooling applications.

Enzyme-catalyzed electrolysis system (EES) is a promising technique for the efficient dechlorination of pollutants. In this study, ionic liquids (ILs) was first introduced to enhance the dichloromethane dechlorination performance of an EES. An imidazole-based IL, 1-ethyl-3-methylimidazole tetrafluoroborate ([EMIM][BF4]), was chosen due to its excellent performance on dechlorination enhancement than other three ILs. The cyclic voltammograms with different scan rates shows that the presence of IL increased the apparent electron transfer rate constant (ks) from 0.008 to 0.013 s−1. The calculated surface electroactive species concentration (τc) also increased from 7.8 × 10−9 to 9.5 × 10−9 mol cm−2. Electrochemical impedance spectroscopy analysis illustrates that the IL mainly weakened the interfacial resistance between electrolyte and cathode to accelerate the electron communication in the EES. The introduction of IL facilitated the regeneration of reduced glutathione from oxidized glutathione, whereas inhibited the catalytic activity of dehalogenase via the disruption of secondary structure shown in circular dichroism spectra. The presence of IL was also facilitated the dichloromethane diffusion from electrolyte to cathode. The mass transfer rate constants of dichloromethane (km,d) increased by 6.9 times after the addition of IL. The optimum volume concentration, pH value, reaction temperature and applied voltage were 20%, 7, 35 °C and −0.8 V vs Ag/AgCl, respectively. The study is helpful to understand the promotion mechanism of IL on the dechlorination performance of EES when it is adopted as a treatment technique.

Hierarchical porous carbons are successfully prepared from a readily water-soluble biomass sodium alginate with the assistance of Fe3+ and Ni2+ via a simple sol–gel method. The drying method and separation ways of ionically-crosslinked alginate gels could significantly affect the morphologies of the precursors and the porous structures for the carbons. By adjusting the amounts of FeCl3·6H2O, we can prepare hierarchical porous carbons and target the surface area and pore volume. The results show that the porous carbons exhibit superior electrochemical behaviors when they are used as anode materials for lithium-ion batteries. Both the porous structures and the assembly way of graphene sheets in the carbon materials play important roles in their electrochemical properties.

Graphitic carbon nitride (g-C3N4), liking a yolk-shell cake, was successfully prepared via a gas-bubble template method with the assistance of NH4Cl. In this special structure, the shell region consists of numerous thin sheets with rich porosity, while the yolk region consists of a lot of cross-linked large hollow particles. The formation mechanism of yolk-shell structure was investigated via adjusting the mass ratio of NH4Cl to melamine, and calcination temperature. The surface area and pore volume of the two types of porous g-C3N4 nanomaterials significantly depend on the mass ratio of NH4Cl to melamine in a certain extent. The as-prepared porous g-C3N4 nanomaterials exhibit improved photocatalytic activity compared to bulk g-C3N4. Moreover, the porous thin sheets in shell region possess larger specific surface area, higher light utilization, higher charge separation and transfer efficiency, than the hollow particles in yolk region, leading to the superior photocatalytic performance. We believe that this simple, economical, and nontoxic approach for the preparation of g-C3N4 nanomaterials opens new possibilities for scalable synthesis of g-C3N4-based materials with more controllable structures.

A simple template-free solvothermal route has been successfully developed to prepare iron-based fluoride nanostructures with controlled morphology and composition. Fe and F sources, reactant concentration, solvent composition and solvothermal reaction time play important roles in the control of the nanostructures and composition of the products. FeF3·0.33H2O hollow nanospheres exhibit high reversible capacities and good cycling performance when used as cathode materials for lithium ion batteries, and the hollow structure has an important impact on the electrochemical performance. In addition, the mesoporous structures within the materials provide an easily accessible system for lithium ion and electrolyte transportation diffusion. The small mesopores in the nanoparticles can also make the electrolyte and lithium ion further diffuse into the interior of the electrode materials and increase the electrolyte/electrode contact area.

Enzymatic electrolysis cell (EEC) has advantages over microbial electrolysis cell (MEC) due to the needless of microbe inoculation and high-efficiency of enzymatic reaction. In this study, an EEC was first applied to achieve the effective degradation of halogenated organic pollutants and dichloromethane (CH2Cl2) was utilized as a model pollutant. The results indicate that the degradation efficiency of CH2Cl2 after 2 hr reaction in the EEC was almost 100%, which was significantly higher than that with enzyme (51.1%) or current (19.0%). The current induced the continuous regeneration of reduced glutathione (GSH), thus CH2Cl2 was degraded under the catalysis of GSH-dependent dehalogenase through stepwise dechlorination, and successively formed monochloromethane (CH3Cl) and methane (CH4). The kinetic result shows that with a current of 15 mA, the maximum specific degradation rate of CH2Cl2 (3.77 × 10−3hr-1) was increased by 5.7 times. The optimum condition for CH2Cl2 dechlorination was also obtained with pH, current and temperature of 7.0, 15 mA and 35°C, respectively. Importantly, this study helps to understand the behavior of enzymes and the fate of halogenated organic pollutants with EEC, providing a possible treatment technology for halogenated organic pollutants.

Malaria is the most devastating parasitic disease worldwide. Artemisinin is the only drug that can cure malaria that is resistant to quinine-derived drugs. After the commercial extraction of artemisinin from Artemisia annua, the recovery of dihydroartemisinic acid (DHAA) from artemisinin extraction by-product has the potential to increase artemisinin commercial yield. Here we describe the development and optimization of an ultrasound-assisted alkaline procedure for the extraction of DHAA from artemisinin production waste using response surface methodology. Our results using this methodology established that NaOH at 0.36%, extraction time of 67.96 min, liquid-solid ratio of 5.89, and ultrasonic power of 83.9 W were the optimal conditions to extract DHAA from artemisinin production waste. Under these optimal conditions, we achieved a DHAA yield of 2.7%. Finally, we conducted a validation experiment, and the results confirmed the prediction generated by the regression model developed in this study. This work provides a novel way to increase the production of artemisinin per cultivated area and to reduce artemisinin production costs by recycling its commercial waste to obtain DHAA, an immediate precursor of artemisinin. The use of this technology may reduce the costs of artemisinin-based antimalarial medicines.

Non-destructive testing is a popular technique for defect assessment of composite materials, where machine learning models become more important in its data analysis. Nevertheless, deep learning, which has achieved state-of-the-art results in many tasks, has received less attention in this field. Herein, a generative independent component (IC) thermography method is proposed. In detail, a generative adversarial network is implemented for image augmentation, which generates fake thermal images that mimic the patterns of real measurements. In doing this, the sample size is enlarged and the defect information contained in the images is enriched. Then, both the real and fake thermal images are decomposed by IC analysis, which separates the defect signals represented by non-Gaussian sources and the non-uniform backgrounds caused by uneven heating. Consequently, the defect detection results are improved. The performance of the proposed method on a polymer composite specimen demonstrates its effectiveness.

Auxin response factors (ARFs) are involved in auxin-mediated transcriptional regulation in plants. In this study, we performed functional characterization of SlARF6A in tomato. SlARF6A is located in the nucleus and exhibits transcriptional activator activity. Overexpression of SlARF6A increased chlorophyll contents in the fruits and leaves of tomato plants, whereas downregulation of SlARF6A decreased chlorophyll contents compared with those of wild-type (WT) plants. Analysis of chloroplasts using transmission electron microscopy indicated increased sizes of chloroplasts in SlARF6A-overexpressing plants and decreased numbers of chloroplasts in SlARF6A-downregulated plants. Overexpression of SlARF6A increased the photosynthesis rate and accumulation of starch and soluble sugars, whereas knockdown of SlARF6A resulted in opposite phenotypes in tomato leaves and fruits. RNA-sequence analysis showed that regulation of SlARF6A expression altered the expression of genes involved in chlorophyll metabolism, photosynthesis and sugar metabolism. SlARF6A directly bound to the promoters of SlGLK1, CAB, and RbcS genes and positively regulated the expression of these genes. Overexpression of SlARF6A also inhibited fruit ripening and ethylene production, whereas downregulation of SlARF6A increased fruit ripening and ethylene production. SlARF6A directly bound to the SAMS1 promoter and negatively regulated SAMS1 expression. Taken together, these results expand our understanding of ARFs with regard to photosynthesis, sugar accumulation and fruit development and provide a potential target for genetic engineering to improve fruit nutrition in horticulture crops.

In order to investigate the physiological functions and biosynthesis regulation of borneol or comphore in Artermisia annua L., which is the major source of the anti-malaria drug artemisinin, the full length cDNA of the gene encoding a borneol dehydrogenase (AaBDH) was cloned from A. annua for the first time by using RACE (rapid amplification of cDNA ends). The completed open read frame of AaBDH was 1415 bp and it encoded a 885-amino acid protein with a predicted molecular mass of 31.04 kDa and a pI of 6.16. AaBDH showed 68–70% of amino acid identity to alcohol dehydrogenases from Solanum lycopersicum, Ppulus trichocarpa, Morus notabili and Ricinus communis. While it shared 51% and 58% of identity with artemisia alcohol dehydrogenase ADH2 from A. annua and borneol dedrogenase LiBDH from Lavandula x intermedia, respectively. The recombinant protein was obtained by heterogeneous expression of AaBDH in a strain of Escherichia coli BL 21 and purified by affinity chromatography. The function of AaBDH was characterized by using of in vitro enzymatic assays, and the results showed that AaBDH had the ability to specifically convert borneol into camphor in the presence of NAD+ (nicotinamide adenine dinucleotide) or NADP+ (nicotinamide adenine dinucleotide phosphate). The cloning of AaBDH laid a significant foundation for further investigation on physiological functions and biosynthesis regulation of plant monoterpenoids.

Canola is an important vegetable oil crop globally, and the understanding of the molecular mechanism underlying fatty acids biosynthesis during seed embryo development is an important research goal. Here we report the transcriptional profiling analysis of developing canola embryos using RNA-sequencing (RNA-Seq) method. RNA-Seq analysis generated 58,579,451 sequence reads aligned with 32,243 genes. It was found that a total of 55 differential expression genes (DEGs) encoding 28 enzymes function in carbon flow to fatty acids of storage TAG. Most of the DEGs encoding above enzymes showed similar expression pattern, indicating the DEGs are cooperatively involved in carbon flow into fatty acids. In addition, 41 DEGs associated with signal transductions, transport and metabolic processing of auxin, gibberellin, abscisic acid, cytokinin and salicylic acids were found in the RNA-Seq database, which indicates the important roles of the phytohormones in controlling embryo development and fatty acids synthesis. 122 DEGs encoding transcriptional factor family members were found in developing canola embryos. Furthermore, BnDOF5.6, a zinc finger transcriptional factor gene, found in RNA-Seq database was down-regulated in developing canola embryos. The transgenic plants displayed reduced embryo sizes, decreased fatty acids contents and altered seed fatty acids composition in canola. Down-regulated of BnDof5.6 also changed the expression levels of genes involved in fatty acids synthesis and desaturation. Our results indicate that BnDof5.6 is required for embryo development and fatty acids synthesis in canola. Overall this study presents new information on the global expression patterns of genes during embryo development and will expand our understanding of the complex molecular mechanism of carbon flow into fatty acids and embryo development in canola.

In order to make full use of artemisinin production waste and thus to reduce the production cost of artemisinin, we developed an efficient and scalable method to isolate high-purity dihydroartemisinic acid from artemisinin production waste by combining anion-exchange resin with silica-gel column chromatography. The adsorption and desorption characteristics of dihydroartemisinic acid on 10 types of anion-exchange resin were investigated, and the results showed that the 717 anion-exchange resin exhibited the highest capacity of adsorption and desorption to dihydroartemisinic acid. Adsorption isotherms were established for the 717 anion-exchange resin and they fitted well with both Langmuir and Freundlich model. Dynamic adsorption and desorption properties of 717 anion-exchange resin were characterized to optimize the chromatographic conditions. Subsequently, the silica-gel column chromatography was performed and dihydroartemisinic acid with a purity of up to 98% (w/w) was obtained. Finally, the scale-up experiments validated the preparative separation of high-purity dihydroartemisinic acid from industrial waste developed in the present work. This work presented for the first time an isolation of dihydroartemisinic acid with a purity of 98% from Artemisia annua (A. annua) by-product, which adds more value to this crop and has the potential to lower the prices of anti-malarial drugs.

This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017.

The regulatory mechanism of flavonoids, which synergise anti-malarial and anti-cancer compounds in Artemisia annua, is still unclear. In this study, an anthocyanidin-accumulating mutant callus was induced from A. annua and comparative transcriptomic analysis of wild-type and mutant calli performed, based on the next-generation Illumina/Solexa sequencing platform and de novo assembly. A total of 82,393 unigenes were obtained and 34,764 unigenes were annotated in the public database. Among these, 87 unigenes were assigned to 14 structural genes involved in the flavonoid biosynthetic pathway and 37 unigenes were assigned to 17 structural genes related to metabolism of flavonoids. More than 30 unigenes were assigned to regulatory genes, including R2R3-MYB, bHLH and WD40, which might regulate flavonoid biosynthesis. A further 29 unigenes encoding flavonoid biosynthetic enzymes or transcription factors were up-regulated in the mutant, while 19 unigenes were down-regulated, compared with the wild type. Expression levels of nine genes involved in the flavonoid pathway were compared using semi-quantitative RT-PCR, and results were consistent with comparative transcriptomic analysis. Finally, a putative flavonol synthase gene (AaFLS1) was identified from enzyme assay in vitro and in vivo through heterogeneous expression, and confirmed comparative transcriptomic analysis of wild-type and mutant callus. The present work has provided important target genes for the regulation of flavonoid biosynthesis in A. annua.

[Research background] Different social forms determine different social governance models. With the development of the Internet of things, cloud computing and artificial intelligence, digital governance has entered the big data era. [Research questions and methods] What should be the digital governance model for the big data era? Krassimira believe that applying digital governance theory to guide smart city practice is an important step in governance theory in the field of urban governance. Singapore's "Smart nation 2025" is the first blueprint for a smart nation in the world. Taking the practice of Singapore's “smart nation 2025” as a case study, based on the theory of digital governance, from the three dimensions of digital economy, digital government and digital society, using the "institutional-organization-technology-behavior" analysis framework, this paper analyzes the context framework of Singapore's digital governance model, and summarizes the typical characteristics of Singapore's digital governance model. [Conclusion] The study found that Singapore has established a digital governance model in which governments, economic society, and civil society can collaborate and interact. Singapore has accumulated a wealth of digital governance experience[Contribution and limitation] Singapore's urban development is highly mature, and its digital governance has typical particularity, but the experience and enlightenment of Singapore's digital governance practice can provide a reference path for improving China's digital governance ability.

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT • Co-administration of gemfibrozil significantly increases the exposure of repaglinide. • CYP3A4 and CYP2C8 are important in the metabolism of repaglinide. • OATP1B1 polymorphism is an independent predictor of repaglinide pharmacokinetics. • Gemfibrozil and its O-glucuronide are CYP2C8 and OATP1B1 inhibitors. WHAT THIS STUDY ADDS • Acyl glucuronidation is a major metabolic pathway of repaglinide in vitro. • Gemfibrozil and its glucuronide inhibit the glucuronidation of repaglinide. • UGT1A1 is a major enzyme responsible for the glucuronidation of repaglinide. AIM To further explore the mechanism underlying the interaction between repaglinide and gemfibrozil, alone or in combination with itraconazole. METHODS Repaglinide metabolism was assessed in vitro (human liver subcellular fractions, fresh human hepatocytes, and recombinant enzymes) and the resulting incubates were analyzed, by liquid chromatography-mass spectrometry (LC-MS) and radioactivity counting, to identify and quantify the different metabolites therein. Chemical inhibitors, in addition to a trapping agent, were also employed to elucidate the importance of each metabolic pathway. Finally, a panel of human liver microsomes (genotyped for UGT1A1*28 allele status) was used to determine the importance of UGT1A1 in the direct glucuronidation of repaglinide. RESULTS The results of the present study demonstrate that repaglinide can undergo direct glucuronidation, a pathway that can possibly contribute to the interaction with gemfibrozil. For example, [3H]-repaglinide formed glucuronide and oxidative metabolites (M2 and M4) when incubated with primary human hepatocytes. Gemfibrozil effectively inhibited (∼78%) both glucuronide and M4 formation, but had a minor effect on M2 formation. Concomitantly, the overall turnover of repaglinide was also inhibited (∼80%), and was completely abolished when gemfibrozil was co-incubated with itraconazole. These observations are in qualitative agreement with the in vivo findings. UGT1A1 plays a significant role in the glucuronidation of repaglinide. In addition, gemfibrozil and its glucuronide inhibit repaglinide glucuronidation and the inhibition by gemfibrozil glucuronide is time-dependent. CONCLUSIONS Inhibition of UGT enzymes, especially UGT1A1, by gemfibrozil and its glucuronide is an additional mechanism to consider when rationalizing the interaction between repaglinide and gemfibrozil.

BMS-690514, a potent inhibitor of human epidermal growth factor receptor (HER) 1 (EGFR), 2, and 4, and vascular endothelial growth factor receptors (VEGFR) 1-3, is currently under investigation as an oral agent for the treatment of solid tumors. In vitro and in vivo studies were conducted to characterize the pharmacokinetics and metabolism. Through integration of in vitro and in vivo pharmacokinetic data and antitumor efficacy in nude mice, human pharmacokinetics and efficacious doses were projected for BMS-690514. The oral bioavailability of BMS-690514 was 78% in mice, approximately 100% in rats, 8% in monkeys, and 29% in dogs. The low oral bioavailability in monkeys could be attributed to high systemic clearance in that species, which was also consistent with predicted clearance using in vitro data from monkey liver microsomes. Permeability of BMS-690514 in Caco-2 cells was in the intermediate range with a moderate potential to be a P-gp substrate. Experiments using recombinant human CYP enzymes and human liver microsomes suggested that CYP2D6 and CYP3A4 are likely to play a key role in the metabolic clearance of BMS-690514; in addition, direct glucuronidation of BMS-690514 was also observed in human hepatocytes. BMS-690514 was able to cross the blood-brain barrier with a brain-to-plasma ratio of approximately 1. The preclinical ADME properties of BMS-690514 suggest good oral bioavailability in humans and metabolism by multiple pathways including oxidation and glucuronidation. Based on the efficacious AUC in nude mice and predicted human pharmacokinetics, the human efficacious QD dose is predicted to be in the range of 100-200 mg.

Dihydroartemisinic acid (DHAA) is the direct precursor to artemisinin, an effective anti-malaria compound from Artemisia annua L. (A. annua), and it can be transformed to artemisinin without the catalysis of enzyme. A rapid and sensitive analysis of DHAA in A. annua is needed to screen excellent plant resources aimed to improve artemisinin production. In order to develop a rapid and sensitive determination method for DHAA in plant, the extraction and analysis conditions were extensively investigated in the present work. As a result, extraction of powdered A. annua leaves at 55°C for 50 min with chloroform resulted in the highest yield of DHAA, with a recovery of98%. The precision of this gas chromatographic procedure ranged from 1.22 to 2.94% for intra-day and from 1.69 to 4.31% for inter-day, respectively. The accuracy was 99.55-103.02% for intra-day and 98.86-99.98% for inter-day, respectively. The measured LOQ and LOD values of the proposed method reached 5.00 and 2.00 μg/mL, respectively. Validation indicated the method was robust, quick, sensitive and adequate for DHAA analysis.

The auxin response factor (ARF) genes encode a large family of proteins involved in auxin signaling transduction. SlARF3, a member of the ARF gene family, encodes a protein containing two conserved domains, B3 and ARF, and lacking an Aux/IAA domain. Expression analysis showed that SlARF3 has a particularly high expression level in trichomes. In situ hybridization also detected the SlARF3 transcripts in epidermal pavement cells of leaves. The physiological function of SlARF3 was studied by using the RNA interference (RNAi) strategy. SlARF3-down-regulated plants exhibited decreased density of epidermal pavement cells and obviously reduced density of type I, V and VI trichomes of leaves, which indicates the important role of SlARF3 in the formation of trichomes and epidermal cells in tomato. The number of shoot xylem cells was also decreased in SlARF3-down-regulated lines. Furthermore, RNA-sequencing (RNA-Seq) analysis identified 51 differentially expressed genes (DEGs) belonging to 14 transcription factor (TF) families, such as MYB, bHLH, WD40 and C2H2 zinc finger. Twenty-seven DEGs were involved in the metabolism and signaling transduction of phytohormones, such as auxin, ethylene and gibberellin. These results indicated the important roles of the TFs and hormones in auxin-dependent transcriptional regulation of trichome formation in tomato. Taken together, our results demonstrate that SlARF3 plays an important role in the formation of epidermal cells and trichomes and reveal novel and specific functions for ARFs in tomato developmental processes.

The development and progression of prostate cancer is known to be dependent on androgens and their signaling mediated by the androgen receptor (AR). The primary therapeutic intervention involves using agents that lower serum testosterone (e.g., LHRH agonists), often in concert with an AR antagonist, such as bicalutamide. Despite a favorable initial anti-tumor response, most patients progress to the advanced hormone-refractory disease. The development of resistance to anti-androgen therapy has been shown to be associated with an increase in the levels of both AR mRNA and protein. This observation supports the concept that an AR antagonist with a significant improvement in potency as compared to bicalutamide and a broader spectrum of in vivo anti-tumor activity, including the bicalutamide-refractory human prostate tumor xenografts, may provide a significant clinical advantage in the treatment of advanced prostate cancer. This presentation will describe structure-activity relationships in a novel tetracyclic series of androgen receptor antagonists leading up to the identification of BMS-779333. It is a potent AR full antagonist, which exhibited broad spectrum efficacy in four human prostate tumor xenograft models. BMS-779333 did not exhibit agonist activity for AR mutant isoforms. Tumors that failed bicalutamide treatment were shown to retain their sensitivity to respond to BMS-779333. Transcriptomic changes in LuCaP-35 tumors treated with BMS-779333 were closer to castration than with other drug treatments. Based on its overall profile, BMS-779333 was selected for further preclinical evaluation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5782.