Ferroptosis MRI for early detection of anticancer drug-induced acute cardiac/kidney injuries.

Ferroptosis has been realized in anticancer drug-induced acute cardiac/kidney injuries (ACI/AKI); however, molecular imaging approach to detect ferroptosis in ACI/AKI is a challenge. We report an artemisinin-based probe (Art-Gd) for contrast-enhanced magnetic resonance imaging of ferroptosis (feMRI) by exploiting the redox-active Fe(II) as a vivid chemical target. In vivo, the Art-Gd probe showed great feasibility in early diagnosis of anticancer drug-induced ACI/AKI, which was at least 24 and 48 hours earlier than the standard clinical assays for assessing ACI and AKI, respectively. Furthermore, the feMRI was able to provide imaging evidence for the different mechanisms of action of ferroptosis-targeted agents, either by blocking lipid peroxidation or depleting iron ions. This study presents a feMRI strategy with simple chemistry and robust efficacy for early evaluation of anticancer drug-induced ACI/AKI, which may shed light on the theranostics of a variety of ferroptosis-related diseases.

Identification and Analysis of Stress-Associated Proteins (SAPs) Protein Family and Drought Tolerance of ZmSAP8 in Transgenic Arabidopsis.

Stress-associated proteins (SAPs), a class of A20/AN1 zinc finger proteins, play vital roles in plant stress response. However, investigation of SAPs in maize has been very limited. Herein, to better trace the evolutionary history of SAPs in maize and plants, 415 SAPs were identified in 33 plant species and four species of other kingdoms. Moreover, gene duplication mode exploration showed whole genome duplication contributed largely to SAP gene expansion in angiosperms. Phylogeny reconstruction was performed with all identified SAPs by the maximum likelihood (ML) method and the SAPs were divided into five clades. SAPs within the same clades showed conserved domain composition. Focusing on maize, nine ZmSAPs were identified. Further promoter cis-elements and stress-induced expression pattern analysis of ZmSAPs indicated that ZmSAP8 was a promising candidate in response to drought stress, which was the only AN1-AN1-C2H2-C2H2 type SAP in maize and belonged to clade I. Additionally, ZmSAP8 was located in the nucleus and had no transactivation activity in yeast. Overexpressing ZmSAP8 enhanced the tolerance to drought stress in Arabidopsis thaliana, with higher seed germination and longer root length. Our results should benefit the further functional characterization of ZmSAPs.

ZmmiR190 and its target regulate plant responses to drought stress through an ABA-dependent pathway.

MicroRNAs (miRNAs) are small, non-coding regulatory RNAs that regulate gene expression by facilitating target mRNA cleavage in plants. They are crucial for responses to diverse stresses. The novel drought-responsive miRNA ZmmiR190 was previously identified during an analysis of the maize transcriptome. In this study, we revealed that transgenic Arabidopsis thaliana overexpressing ZmmiR190 is more sensitive to drought than the wild-type control. The transcript of a nuclear-localized gene, ZmCRP04, was identified as a likely target of ZmmiR190. Moreover, ZmmiR190 and ZmCRP04 had the opposite expression profiles following drought and salt treatments. Additionally, 5' RACE and coexpression analyses in A. thaliana provided evidence of the in vivo targeting of the ZmCRP04 transcript by ZmmiR190. Furthermore, the overexpression of ZmCRP04 in A. thaliana and rice significantly enhanced drought tolerance, with lower malonaldehyde contents and relative electrolyte leakage in the transgenic A. thaliana and rice plants than in the wild-type control. Transgenic plants overexpressing ZmmiR190 or ZmCRP04 were hypersensitive to abscisic acid. These results suggest that the ZmCRP04 transcript is targeted by ZmmiR190 and may encode a protein that positively regulates drought stress tolerance via an abscisic acid-dependent pathway. These findings may be relevant for future molecular breeding aimed at improving crop drought tolerance.

Developing rapid growing Bacillus subtilis for improved biochemical and recombinant protein production.

Bacillus subtilis is a model Gram-positive bacterium, which has been widely used as industrially important chassis in synthetic biology and metabolic engineering. Rapid growth of chassis is beneficial for shortening the fermentation period and enhancing production of target product. However, engineered B. subtilis with faster growth phenotype is lacking. Here, fast-growing B. subtilis were constructed through rational gene knockout and adaptive laboratory evolution using wild type strain B. subtilis 168 (BS168) as starting strain. Specifically, strains BS01, BS02, and BS03 were obtained through gene knockout of oppD, hag, and flgD genes, respectively, resulting 15.37%, 24.18% and 36.46% increases of specific growth rate compared with BS168. Next, strains A28 and A40 were obtained through adaptive laboratory evolution, whose specific growth rates increased by 39.88% and 43.53% compared to BS168, respectively. Then these two methods were combined via deleting oppD, hag, and flgD genes respectively on the basis of evolved strain A40, yielding strain A4003 with further 7.76% increase of specific growth rate, reaching 0.75 h-1 in chemical defined M9 medium. Finally, bioproduction efficiency of intracellular product (ribonucleic acid, RNA), extracellular product (acetoin), and recombinant proteins (green fluorescent protein (GFP) and ovalbumin) by fast-growing strain A4003 was tested. And the production of RNA, acetoin, GFP, and ovalbumin increased 38.09%, 5.40%, 9.47% and 19.79% using fast-growing strain A4003 as chassis compared with BS168, respectively. The developed fast-growing B. subtilis strains and strategies used for developing these strains should be useful for improving bioproduction efficiency and constructing other industrially important bacterium with faster growth phenotype.

Towards next-generation model microorganism chassis for biomanufacturing.

Synthetic biology provides powerful tools and novel strategies for designing and modifying microorganisms to function as cell factories for biomanufacturing, which is a promising approach for realizing chemical production in a green and sustainable manner. Recent advances in genetic component design and genome engineering have enabled significant progresses in the field of synthetic biology chassis that have been developed for enzymes or biochemical production based on synthetic biology strategies, with particular reference to model microorganisms, such as Escherichia coli, Bacillus subtilis, Corynebacterium glutamicum, and Saccharomyces cerevisiae. In this review, strategies for engineering four different functional cellular modules which encompass the total process of biomanufacturing are discussed, including expanding the substrate spectrum for substrate uptake modules, refactoring biosynthetic pathways and dynamic regulation for product synthesis modules, balancing energy and redox modules, and cell membrane and cell wall engineering of product storage and secretion modules. Novel strategies of integrating and coordinating different cellular modules aided by synthetic co-culturing of multiple chassis, artificial intelligence-aided data mining for guiding strain development, and the process for designing automatic chassis development via biofoundry are expected to generate next generations of model microorganism chassis for more efficient biomanufacturing. KEY POINTS: • Engineering of functional cellular modules facilitate next generations of chassis construction. • Global optimization of biosynthesis can be improved by metabolic models. • Data-driven and automatic strain development can improve microorganism chassis construction.

Enhanced removal of Cr(VI) by silicon rich biochar-supported nanoscale zero-valent iron.

Silicon-rich biochar-supported nanoscale zero-valent iron (nZVI) was studied to evaluate enhanced removal of hexavalent chromium (Cr(VI)) in solution. The compositional structures of the nZVI and biochar-supported nZVI were analyzed by Fourier transform infrared spectroscopy, X-ray diffraction and X-ray photoelectron spectra before and after Cr(VI) reaction. The removal amount of Cr(VI) by nZVI-RS700 (rice straw pyrolyzed at 700 °C) was considerably greater than that by nZVI and other biochar-supported nZVI samples. Upon the silicon was removed from RS700 (nZVI-RS700(-Si)), a significant decreased removal of Cr(VI) was observed. It was revealed that nZVI supported by silicate particles of biochar and the promotion of iron oxidation by SiO2 both contribute to the enhanced Cr(VI) removal. We found that the reduction and adsorption both contributed to the removal of Cr(VI), ferrous chromite (FeCr2O4) was observed on the surface of the nZVI-RS700 composite. The formation of FeCr2O4 is attributed to the reduction of Cr(VI) by nZVI and the adsorption of chromium oxide with iron on the surface of RS700. Therefore, RS700-supported nZVI can be used as a potential remediation reagent to treat Cr(VI)-contaminated groundwater.

Effect of culturing temperatures on cadmium phytotoxicity alleviation by biochar.

Biochar produced from rice straw at 400 °C (RS400) was prepared to determine its alleviating effect on Cd phytotoxicity to wheat seedlings under different cultivation temperatures and pH. A hydroponic system (pH 4.3) and a loam soil slurry system were designed to respectively simulate acidic and neutral soil condition, and cultivation at increasing temperatures (20, 25, and 30 °C) were performed to evaluate the greenhouse effect. The root and shoot elongation and the Cd concentration in root and solution were measured; furthermore, batch experiments for Cd adsorption were undertaken. An increasing inhibition of the root by Cd addition was observed at increasing temperatures. The inhibition rate was 50.50 and 20.80% in hydroponic system and slurry system at 25 °C, respectively; however, the corresponding inhibition rates of root were significantly decreased to 25.5 and 3.5% with addition of RS400. This is mainly attributed to the reduction of Cd migration into the roots by RS400, which decreased Cd bioavailability. The mechanism behind the reduced Cd bioavailability is attributed to the Cd adsorption and the strong buffering capacity of acidity by RS400. Therefore, biochar could be a potential amendment for the remediation of Cd-contaminated soil even at increasing culturing temperatures.

Degradation of 1,4-dioxane by biochar supported nano magnetite particles activating persulfate.

Nano magnetite biochar composite (nFe3O4/biochar) was synthesized and used to activate persulfate for the degradation of 1,4-dioxane. Analytical techniques using X-ray diffraction (XRD), fourier transform infrared analysis (FTIR), X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) indicated that nFe3O4 was spherical and successfully loaded onto the surface of biochar. The results of batch-scale experiments illustrated that the 1,4-dioxane degradation efficiency in aqueous phase was 98.0% after 120 min reaction with the composite mass ratio of 1:1 between nFe3O4 and the pine needle biochar pyrolyzed at 400 °C (P400) under the initial neutral pH. An electron paramagnetic resonance (EPR) study, free radical quenching experiment and XPS analysis were undertaken to illustrate the mechanism of persulfate activation by nFe3O4/biochar. Under acidic and neutral conditions, the predominant free radical was SO4- whereas OH and SO4- predominated when the initial pH was 9.0. The XPS analysis indicated that Fe(II) and oxygenated functional groups activated persulfate. In addition, carbon-carbon double bonds would be transformed into ketone and quinone which could activate persulfate during the reaction.