Bimetallic Cu@Ru Core-Shell Structures with Ligand Effects for Endo-Exogenous Stimulation-Mediated Dynamic Oncotherapy.

Dynamic therapies, which induce reactive oxygen species (ROS) production in situ through endogenous and exogenous stimulation, are emerging as attractive options for tumor treatment. However, the complexity of the tumor substantially limits the efficacy of individual stimulus-triggered dynamic therapy. Herein, bimetallic copper and ruthenium (Cu@Ru) core-shell nanoparticles are applied for endo-exogenous stimulation-triggered dynamic therapy. The electronic structure of Cu@Ru is regulated through the ligand effects to improve the adsorption level for small molecules, such as water and oxygen. The core-shell heterojunction interface can rapidly separate electron-hole pairs generated by ultrasound and light stimulation, which initiate reactions with adsorbed small molecules, thus enhancing ROS generation. This synergistically complements tumor treatment together with ROS from endogenous stimulation. In vitro and in vivo experiments demonstrate that Cu@Ru nanoparticles can induce tumor cell apoptosis and ferroptosis through generated ROS. This study provides a new paradigm for endo-exogenous stimulation-based synergistic tumor treatment.

Identification and characteristics of a novel gland-forming gene in cotton.

The cotton (Gossypium hirsutum) pigment gland is a distinctive structure that functions as the main deposit organ of gossypol and its derivatives. It is also an ideal system in which to study cell differentiation and organogenesis. However, only a few genes that determine the process of gland formation have been reported, including GoPGF, CGP1, and CGFs; the molecular mechanisms underlying gland initiation are still largely unclear. Here, we report the discovery of the novel stem pigment gland-forming gene GoSPGF by map-based cloning; annotated as a GRAS transcription factor, this gene is responsible for the glandless trait specifically on the stem. In the stem glandless mutant T582, a point mutation (C to A) was found to create a premature stop codon and truncate the protein. Similarly, virus-induced gene silencing of GoSPGF resulted in glandless stems and dramatically reduced gossypol content. Comparative transcriptomic data showed that loss of GoSPGF significantly suppressed expression of many genes involved in gossypol biosynthesis and altered expression of genes involved in gibberellic acid signaling/biosynthesis. Overall, these findings provide more insight into the networks regulating glandular structure differentiation and formation in cotton, which will be helpful for understanding other plants bearing special gland structures such as tobacco (Nicotiana benthamiana), artemisia annua, mint (Mentha spp.), and rubber (Hevea brasiliensis).

Highly efficient ammonium removal through nitrogen assimilation by a hydrogen-oxidizing bacterium, Ideonella sp. TH17.

Ideonella sp. TH17, an autotrophic hydrogen-oxidizing bacterium (HOB), was successfully enriched and isolated from activated sludge in a domestic wastewater treatment plant (WWTP). Batch experiments were conducted to identify the cell growth and ammonium (NH4+-N) removal, and to verify the pathways of nitrogen utilization under different conditions. At a representative NH4+-N concentration of 100 mg/L in domestic wastewater, it was the first time that a HOB strain achieved a nearly 100% ammonium removal. More than 90% of NH4+-N was assimilated to biomass nitrogen by strain TH17. Only a little of N2 (<10% of initial NH4+-N) was detected without N2O emission in aerobic denitrification process. Autotrophic NH4+-N assimilation contributed predominantly to biomass nitrogen production, supplemented by assimilatory nitrate (NO3--N) reduction under aerobic conditions. A total of 17 amino acids, accounting for 54.25 ± 1.98% of the dry biomass, were detected in the bacterial biomass harvested at 72 h. These results demonstrated that the newly isolated strain TH17 was capable of removing NH4+-N and recovering nutrients from wastewater efficiently. A new solution was thus provided by this HOB strain for ammonium treatment in sustainable WWTPs of future.

Next-generation transgenic cotton: pyramiding RNAi and Bt counters insect resistance.

Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are extensively cultivated worldwide. To counter rapidly increasing pest resistance to crops that produce single Bt toxins, transgenic plant 'pyramids' producing two or more Bt toxins that kill the same pest have been widely adopted. However, cross-resistance and antagonism between Bt toxins limit the sustainability of this approach. Here we describe development and testing of the first pyramids of cotton combining protection from a Bt toxin and RNA interference (RNAi). We developed two types of transgenic cotton plants producing double-stranded RNA (dsRNA) from the global lepidopteran pest Helicoverpa armigera designed to interfere with its metabolism of juvenile hormone (JH). We focused on suppression of JH acid methyltransferase (JHAMT), which is crucial for JH synthesis, and JH-binding protein (JHBP), which transports JH to organs. In 2015 and 2016, we tested larvae from a Bt-resistant strain and a related susceptible strain of H. armigera on seven types of cotton: two controls, Bt cotton, two types of RNAi cotton (targeting JHAMT or JHBP) and two pyramids (Bt cotton plus each type of RNAi). Both types of RNAi cotton were effective against Bt-resistant insects. Bt cotton and RNAi acted independently against the susceptible strain. In computer simulations of conditions in northern China, where millions of farmers grow Bt cotton as well as abundant non-transgenic host plants of H. armigera, pyramided cotton combining a Bt toxin and RNAi substantially delayed resistance relative to using Bt cotton alone.

An integrated meta-omics approach reveals the different response mechanisms of two anammox bacteria towards fluoroquinolone antibiotics.

The emerging fluoroquinolone antibiotics (FQs) are highly influential in nitrogen removal from livestock wastewater. However, beyond the capability of nitrogen removal, little is known about the molecular mechanisms (e.g., shift of core metabolism and energy allocation) of different anaerobic ammonium-oxidizing bacteria (AnAOB) under continuous FQ stress. This study investigated the effects of ciprofloxacin, ofloxacin and their mixture at concentrations detected in livestock wastewater on two key anammox species in membrane bioreactors. It was found 20 µg/L FQs promoted nitrogen removal efficiency and community stability, and42-51 % of FQs were removed simultaneously. Integrated meta-omics analysis revealed varied gene expression patterns between the two dominant AnAOB, Candidatus Brocadia sapporoensis (B AnAOB) and Candidatus Kuenenia stuttgartiensis (K AnAOB). The nitrogen metabolic processes were bolstered in B AnAOB, while those involved in anammox pathway of K AnAOB were inhibited. This difference was tentatively attributed to the up-regulation of reactive oxygen species scavenger genes (ccp and dxf) and FQ resistance gene (qnrB72) in B AnAOB. Importantly, most enhanced core biosynthesis/metabolism of AnAOB and close cross-feeding with accompanying bacteria were also likely to contribute to their higher levels of biomass yield and metabolism activity under FQ stress. This finding suggests that B AnAOB has the advantage of higher nitrogen metabolism capacity over K AnAOB in livestock wastewater containing FQs, which is helpful for efficient and stable nitrogen removal by the functional anammox species.

Molecular insights into enhanced nitrogen removal induced by trace fluoroquinolone antibiotics in an anammox system.

It has been widely reported that fluoroquinolones (FQs) can affect the anaerobic ammonium oxidization (anammox) microorganisms, which interferes with the performance of nitrogen removal from wastewater. However, the metabolic mechanism of anammox microorganisms responding to FQs has rarely been explored. In this study, it was found that 20 µg/L FQs promoted the nitrogen removal performance of anammox microorganisms in batch exposure assays, and 36-51% of FQs were removed simultaneously. Combined metabolomics and genome-resolved metagenomic analysis revealed up-regulated carbon fixation in anammox bacteria (AnAOB), while purine and pyrimidine metabolism, protein generation and transmembrane transport were enhanced in AnAOB and symbiotic bacteria by 20 µg/L FQs. Consequently, hydrazine dehydrogenation, nitrite reduction, and ammonium assimilation were bolstered, improving the nitrogen removal efficiency of the anammox system. These results revealed the potential roles of specific microorganisms in response to emerging FQs and provided further information for practical application of anammox technology in wastewater treatment.

Development of the engineered "glanded plant and glandless seed" cotton.

After fiber, cottonseed is the second most important by-product of cotton production. However, high concentrations of toxic free gossypol deposited in the glands of the cottonseed greatly hamper its effective usage as food or feed. Here, we developed a cotton line with edible cottonseed by specifically silencing the endogenous expression of GoPGF in the seeds, which led to a glandless phenotype with an ultra-low gossypol content in the seeds and nearly normal gossypol in other parts of the plants. This engineered cotton maintains normal resistance to insect pests, but the gossypol content in the seeds dropped by 98%, and thus, it can be consumed directly as food. The trait of a low gossypol content in the cottonseeds was stable and heritable, while the protein, oil content, and fiber yield or quality were nearly unchanged compared to the transgenic receptor W0. In addition, comparative transcriptome analysis showed that down-regulated genes in the ovules of the glandless cotton were enriched in terpenoid biosynthesis, indicating the underlying relationship between gland formation and gossypol biosynthesis. These results pave the way for the comprehensive utilization of cotton as a fiber, oil, and feed crop in the future.

Sustainable treatment of nitrate-containing wastewater by an autotrophic hydrogen-oxidizing bacterium.

Bacteria are key denitrifiers in the reduction of nitrate (NO3 --N), which is a contaminant in wastewater treatment plants (WWTPs). They can also produce carbon dioxide (CO2) and nitrous oxide (N2O). In this study, the autotrophic hydrogen-oxidizing bacterium Rhodoblastus sp. TH20 was isolated for sustainable treatment of NO3 --N in wastewater. Efficient removal of NO3 --N and recovery of biomass nitrogen were achieved. Up to 99% of NO3 --N was removed without accumulation of nitrite and N2O, consuming CO2 of 3.25 mol for each mole of NO3 --N removed. The overall removal rate of NO3 --N reached 1.1 mg L-1 h-1 with a biomass content of approximately 0.71 g L-1 within 72 h. TH20 participated in NO3 --N assimilation and aerobic denitrification. Results from 15N-labeled-nitrate test indicated that removed NO3 --N was assimilated into organic nitrogen, showing an assimilation efficiency of 58%. Seventeen amino acids were detected, accounting for 43% of the biomass. Nitrogen loss through aerobic denitrification was only approximately 42% of total nitrogen. This study suggests that TH20 can be applied in WWTP facilities for water purification and production of valuable biomass to mitigate CO2 and N2O emissions.

Ecological and toxicological assessments of anthropogenic contaminants based on environmental metabolomics.

There has long been a great concern with growing anthropogenic contaminants and their ecological and toxicological effects on living organisms and the surrounding environment for decades. Metabolomics, a functional readout of cellular activity, can capture organismal responses to various contaminant-related stressors, acquiring direct signatures to illustrate the environmental behaviours of anthropogenic contaminants better. This review entails the application of metabolomics to profile metabolic responses of environmental organisms, e.g. animals (rodents, fish, crustacean and earthworms) and microorganisms (bacteria, yeast and microalgae) to different anthropogenic contaminants, including heavy metals, nanomaterials, pesticides, pharmaceutical and personal products, persistent organic pollutants, and assesses their ecotoxicological impacts with regard to literature published in the recent five years. Contaminant-induced metabolism alteration and up/down-regulation of metabolic pathways are revealed in typical organisms. The obtained insights of variations in global metabolism provide a distinct understanding of how anthropogenic contaminants exert influences on specific metabolic pathways on living organisms. Thus with a novel ecotechnique of environmental metabolomics, risk assessments of anthropogenic contaminants are profoundly demonstrated.

One-step dynamic fabrication of asymmetric multi-layered porous films for sustained drug delivery.

Asymmetric multi-layered porous films were prepared by casting inverse emulsion following the breath figure method. The porous morphologies both on the surface and in the bulk of the fabricated film could be dynamically manipulated by tuning the emulsion composition as well as the environmental conditions. The model drug was efficiently loaded into the porous film by direct encapsulation during film fabrication, and remarkable sustained drug release from the porous film for more than 28 days was achieved.

Mercury methylation by Geobacter metallireducens GS-15 in the presence of Skeletonema costatum.

In this study, bacterial mercury (Hg) methylation was investigated under the influence of red-tide algae of Skeletonema costatum (S. costatum). The distribution and speciation of total mercury (THg) and methylmercury (MeHg) were profiled by employing Geobacter metallireducens (G. metallireducens GS-15) as the methylating bacteria. G. metallireducens GS-15 showed different capabilities in methylating different inorganic forms of Hg(II) (HgCl2) and Hg(II)-Algae (HgCl2 captured by S. costatum) to MeHg. In the absence of S. costatum, a maximum methylation efficiency of 4.31 ±â€¯0.47% was achieved with Hg(II) of 1-100 µg L-1, while accelerated MeHg formation rate was detected at a higher initial Hg(II) concentration. In the presence of S. costatum, there were distinct changes in the distribution of THg and MeHg by altering the bioavailability of Hg(II) and Hg(II)-Algae. A larger proportion of THg tended to be retained by a higher algal biomass, resulting in decreased methylation efficiencies. The methylation efficiency of Hg(II) decreased from 3.01 ±â€¯0.10% to 1.01 ±â€¯0.01% with 10-mL and 250-mL algal media, and that of Hg(II)-Algae decreased from 0.83 ±â€¯0.13% to 0.22 ±â€¯0.01% with 10-mL and 250-mL Hg(II)-Algae media. Fourier transform infrared spectrometry, surface charge properties and elemental compositions of S. costatum were used to infer that amine, carboxyl and sulfonate functional groups were most likely to interact with Hg(II) through complexation and/or electrostatic attraction. These results suggest that red-tide algae may be an influencing factor on bacterial Hg methylation in eutrophic water bodies.

Inhibitory effects of Skeletonema costatum on mercury methylation by Geobacter sulfurreducens PCA.

Algae and mercury (Hg) are ubiquitous in marine environments. In this study, we investigated the effects of a typical marine algae of diatom Skeletonema costatum on Hg methylation by an iron-reducing bacterium of Geobacter sulfurreducens (G. sulfurreducens) PCA. In the absence of Skeletonema costatum, the bacterial MeHg production rate maximized at 104.06 ±â€¯11.7 ng L-1 h-1 with a high Hg level, while the highest methylation efficiency was achieved at a low Hg concentration. The existence of Skeletonema costatum greatly inhibited the capability of G. sulfurreducens PCA to methylate Hg. With the increase in algal biomass, there was a significant mitigation of MeHg formation and Hg0 release, leaving a considerable proportion of immobilized Hg2+ species (up to 47%) associated with algal cell materials. These results suggest that marine algae are crucial in determining the bioavailability of Hg contaminants and the methylating potential of G. sulfurreducens PCA.

A novel rice C2H2-type zinc finger protein lacking DLN-box/EAR-motif plays a role in salt tolerance.

A cDNA for the gene ZFP182, encoding a C2H2-type zinc finger protein, was cloned from rice by RT-PCR. ZFP182 codes an 18.2 kDa protein with two C2H2-type zinc finger motifs, one nuclear localization signal and one Leu-rich domain. The DLN-box/EAR-motif, which exists in most of plant C2H2-type zinc finger proteins, does not exist in ZFP182. The expression analysis showed that ZFP182 gene was constitutively expressed in leaves, culms, roots and spikes at the adult rice plants, and markedly induced in the seedlings by cold (4 degrees C), 150 mM NaCl and 0.1 mM ABA treatments. The approximate 1.4 kb promoter region of ZFP182 gene was fused into GUS reporter gene and transformed into tobacco. The histochemical analysis revealed that GUS expression could not be detected in transformed tobacco seedlings under normal conditions, but strongly observed in tobacco leaf discs and the vascular tissue of roots treated with NaCl or KCl. Expression of ZFP182 in transgenic tobacco and overexpression in rice increased plant tolerance to salt stress. These results demonstrated that ZFP182 might be involved in plant responses to salt stress.

Distribution and speciation of mercury affected by humic acid in mariculture sites at the Pearl River estuary.

At the Pearl River Estuary of southern China, mercury and its environmental problems have long been a great concern. This study investigated the distribution and speciation of mercury compounds that are significantly influenced by the increasing content of humic acid (HA, a model natural organic matter) in this region. The inorganic mercury and methyl mercury, being adsorbed and converted at different HA levels, were studied in sediments and surface water at both mariculture and their reference sites. In mariculture sediments with higher HA content (up to 4.5%), more mercury were adsorbed at different compound levels, promoting the methylation and accumulation of mercury (P < 0.05) at the sediment-water interface. Seasonal shift in environmental temperature might control the HA content, subsequently favouring mercury methylation (maximum 1.75 ±â€¯0.08 mg L-1 d-1) under warm weather conditions. In reference sites received less HA wastes, lower adsorption capacity and methylation rate were observed for mercury in sediments and surface water. Our work points to the significant roles of HA on mercury distribution and speciation both spatially and seasonally, thus addressing the impacts of mariculture activities on estuary eco-system.

Ubiquity of polystyrene digestion and biodegradation within yellow mealworms, larvae of Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae).

Academics researchers and "citizen scientists" from 22 countries confirmed that yellow mealworms, the larvae of Tenebrio molitor Linnaeus, can survive by eating polystyrene (PS) foam. More detailed assessments of this capability for mealworms were carried out by12 sources: five from the USA, six from China, and one from Northern Ireland. All of these mealworms digested PS foam. PS mass decreased and depolymerization was observed, with appearance of lower molecular weight residuals and functional groups indicative of oxidative transformations in extracts from the frass (insect excrement). An addition of gentamycin (30 mg g-1), a bactericidal antibiotic, inhibited depolymerization, implicating the gut microbiome in the biodegradation process. Microbial community analyses demonstrated significant taxonomic shifts for mealworms fed diets of PS plus bran and PS alone. The results indicate that mealworms from diverse locations eat and metabolize PS and support the hypothesis that this capacity is independent of the geographic origin of the mealworms, and is likely ubiquitous to members of this species.

Fabrication of Active Surfaces with Metastable Microgel Layers Formed during Breath Figure Templating.

Patterned porous surfaces with responsive functionalities are fabricated by a thermoresponsive microgel-assisted breath figure (BF) process. When water droplets submerge into a polystyrene (PS) solution during formation of a porous surface by the bottom-up BF process, poly(N-isopropylacrylamide)-co-acrylic acid (PNIPAm-co-AA) microgels dispersed in the solution spontaneously assemble at the water-organic interfaces like "Pickering emulsions", reinforced by capillary flow. The conformal layer of PNIPAm-co-AA microgels lining the pores appears in images from a scanning electron microscope (SEM) either as a smooth surface layer (L) or as an array of domelike protrusions (D), depending on the conditions at which the sample was dried for SEM. The change between L and D morphology correlates with the volume phase transition behavior of the microgels freely suspended: drying at a temperature below the volume phase transition temperature (VPTT) gives L, and the D morphology is formed by drying at a temperature greater than the VPTT of PNIPAm-co-AA microgels. The morphological transition is shown to accompany a significant change in surface contact angle (CA) relative to a corresponding pore layer made of PS, with L having a CA that is reduced by 85° relative to PS, while the decrease is only 22° for D. Porous structures with morphologically responsive surfaces could find application in biocatalysis or tissue engineering, for example, with functional enzymes sequestered when microgels are collaped and accessible when the microgels are swollen.

Constitutive expression of a novel antimicrobial protein, Hcm1, confers resistance to both Verticillium and Fusarium wilts in cotton.

Fusarium and Verticillium wilts, two of the most important diseases in cotton, pose serious threats to cotton production. Here we introduced a novel antimicrobial protein Hcm1, which comprised harpin protein from Xanthomonas oryzae pv. oryzicola (Xoc), and the chimeric protein, cecropin A-melittin, into cotton. The transgenic cotton lines with stable Hcm1 expression showed a higher resistance to Verticillium and Fusarium wilts both in greenhouse and field trials compared to controls. Hcm1 enabled the transgenic cotton to produced a microscopic hypersensitive response (micro-HR), reactive oxygen species (ROS) burst, and caused the activation of pathogenesis-related (PR) genes in response to biotic stress, indicating that the transgenic cotton was in a primed state and ready to protect the host from pathogenic infection. Simultaneously, Hcm1 protein inhibited the growth of Verticillium dahliae (V. dahliae) and Fusarium oxysporum (F. oxysporum) in vitro. The spread of fungal biomass was also inhibited in vivo since the V. dahliae biomass was decreased dramatically in transgenic cotton plants after inoculation with V. dahliae. Together, these results demonstrate that Hcm1 could activate innate immunity and inhibit the growth of V. dahliae and F. oxysporum to protect cotton against Verticillium and Fusarium wilts.