Plant immunity suppression by an ß-1,3-glucanase of the maize anthracnose pathogen Colletotrichum graminicola.

BACKGROUND: Many phytopathogens secrete a large number of cell wall degrading enzymes (CWDEs) to decompose host cell walls in order to penetrate the host, obtain nutrients and accelerate colonization. There is a wide variety of CWDEs produced by plant pathogens, including glycoside hydrolases (GHs), which determine the virulence, pathogenicity, and host specificity of phytopathogens. The specific molecular mechanisms by which pathogens suppress host immunity remain obscure. RESULT: In this study, we found that CgEC124 encodes a glycosyl hydrolase with a signal peptide and a conserved Glyco_hydro_cc domain which belongs to glycoside hydrolase 128 family. The expression of CgEC124 was significantly induced in the early stage of Colletotrichum graminicola infection, especially at 12 hpi. Furthermore, CgEC124 positively regulated the pathogenicity, but it did not impact the vegetative growth of mycelia. Ecotopic transient expression of CgEC124 decreased the disease resistance and callose deposition in maize. Moreover, CgEC124 exhibited the ß-1,3-glucanase activity and suppresses glucan-induced ROS burst in maize leaves. CONCLUSIONS: Our results indicate that CgEC124 is required for full virulence of C. graminicola but not for vegetative growth. CgEC124 increases maize susceptibility by inhibiting host reactive oxygen species burst as well as callose deposition. Meanwhile, our data suggests that CgEC124 explores its ß-1,3-glucanase activity to prevent induction of host defenses.

Discovery of lupus nephritis targeted inhibitors based on De novo molecular design: comprehensive application of vinardo scoring, ADMET analysis, and molecular dynamics simulation.

Lupus Nephritis (LN) is an autoimmune disease affecting the kidneys, and conventional drug studies have limitations due to its imprecise and complex pathogenesis. Therefore, the aim of this study was to design a novel Lupus Nephritis-targeted drug with good clinical due potential, high potency and selectivity by computer-assisted approach.NIK belongs to the serine/threonine protein kinase, which is gaining attention as a drug target for Lupus Nephritis. we used bioinformatics, homology modelling and sequence comparison analysis, small molecule ab initio design, ADMET analysis, molecular docking, molecular dynamics simulation, and MM/PBSA analysis to design and explore the selectivity and efficiency of a novel Lupus Nephritis-targeting drug, ClImYnib, and a classical NIK inhibitor, NIK SMI1. We used bioinformatics techniques to determine the correlation between lupus nephritis and the NF-κB signaling pathway. De novo drugs design was used to create a NIK-targeted inhibitor, ClImYnib, with lower toxicity, after which we used molecular dynamics to simulate NIK SMI1 against ClImYnib, and the simulation results showed that ClImYnib had better selectivity and efficiency. Our research delves into the molecular mechanism of protein ligands, and we have designed and validated an excellent NIK inhibitor using multiple computational simulation methods. More importantly, it provides an idea of target designing small molecules.Communicated by Ramaswamy H. Sarma.

Multi-scale characterization and analysis of cellular viscoelastic mechanical phenotypes by atomic force microscopy.

The viscoelasticity of cells serves as a biomarker that reveals changes induced by malignant transformation, which aids the cytological examinations. However, differences in the measurement methods and parameters have prevented the consistent and effective characterization of the viscoelastic phenotype of cells. To address this issue, nanomechanical indentation experiments were conducted using an atomic force microscope (AFM). Multiple indentation methods were applied, and the indentation parameters were gradually varied to measure the viscoelasticity of normal liver cells and cancerous liver cells to create a database. This database was employed to train machine-learning algorithms in order to analyze the differences in the viscoelasticity of different types of cells and as well as to identify the optimal measurement methods and parameters. These findings indicated that the measurement speed significantly influenced viscoelasticity and that the classification difference between the two cell types was most evident at 5 µm/s. In addition, the precision and the area under the receiver operating characteristic curve were comparatively analyzed for various widely employed machine-learning algorithms. Unlike previous studies, this research validated the effectiveness of measurement parameters and methods with the assistance of machine-learning algorithms. Furthermore, the results confirmed that the viscoelasticity obtained from the multiparameter indentation measurement could be effectively used for cell classification. RESEARCH HIGHLIGHTS: This study aimed to analyze the viscoelasticity of liver cancer cells and liver cells. Different nano-indentation methods and parameters were used to measure the viscoelasticity of the two kinds of cells. The neural network algorithm was used to reverse analyze the dataset, and the methods and parameters for accurate classification and identification of cells are successfully found.

Insights into Phakopsora pachyrhizi Effector-Effector Interactions.

The multifaceted role of pathogen-encoded effectors in plant-pathogen interactions is complex and not fully understood. Effectors operate within intricate host environments, interacting with host proteins and other effectors to modulate virulence. The complex interplay between effectors raises the concept of metaeffectors, wherein some effectors regulate the activity of others. While previous research has demonstrated the importance of effector repertoires in pathogen virulence, only a limited number of studies have investigated the interactions between these effectors. This study explores the interactions among Phakopsora pachyrhizi effector candidates (PpECs). P. pachyrhizi haustorial transcriptome analysis identified a collection of predicted PpECs. Among these, PpEC23 was found to interact with PpEC48, prompting further exploration into their potential interaction with other effectors. Here, we utilized a yeast two-hybrid screen to explore protein-protein interactions between PpECs. A split-luciferase complementation assay also demonstrated that these interactions could occur within soybean cells. Interestingly, PpEC48 displayed the ability to interact with several small cysteine-rich proteins (SCRPs), suggesting its affinity for this specific class of effectors. We show that these interactions involve a histidine-rich domain within PpEC48, emphasizing the significance of structural motifs in mediating effector interactions. The unique nature of PpEC48, showing no sequence matches in other organisms, suggests its relatively recent evolution and potential orphan gene status. Our work reveals insights into the intricate network of interactions among P. pachyrhizi effector-effector interactions. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Clinical Significance of Contrast-Enhanced Ultrasound Galactography in Pre-operative Diagnosis of Patients With Pathologic Nipple Discharge.

OBJECTIVE: The aim of the work described here was to investigate the feasibility and diagnostic value of using contrast-enhanced ultrasound (CEUS) galactography with SonoVue in patients with pathologic nipple discharge (PND). METHODS: Twenty-eight patients who underwent breast surgery for PND from May 2019 to August 2021 were included. Routine ultrasound, ductoscopy and CEUS galactography were performed successively. Lesions were diagnosed and localized. The sensitivity, specificity and pre-operative localization value of each examination method were evaluated on post-operative pathology. RESULTS: CEUS galactography was successfully conducted in all 28 patients and revealed negative ductal ectasia, filling stop and filling defect. Ductoscopy revealed positive nodules in 21 cases and negative nodules in 7 cases. A total of 18 nodules were found by routine ultrasound, and the relationship between all nodules and the discharge duct was confirmed after CEUS galactography. Compared with the other two methods, CEUS galactography had higher sensitivity, positive predictive value and negative predictive value (100%, 81.82% and 100%, respectively), while it has the same specificity as routine ultrasound (both 60%). The pre-operative location of the nipple duct was consistent with the intra-operative findings in 28 patients after CEUS galactography. CONCLUSION: The ultrasound contrast agent SonoVue can be used for CEUS galactography in patients with PND. CEUS galactography can improve the detection of ductal nodules and locate the nipple discharge duct pre-operatively. As the technique does not emit radiation and SonoVue is easily metabolized and safe, CEUS galactography is better than conventional imaging for PND patients.

Development of a tongue image-based machine learning tool for the diagnosis of gastric cancer: a prospective multicentre clinical cohort study.

Background: Tongue images (the colour, size and shape of the tongue and the colour, thickness and moisture content of the tongue coating), reflecting the health state of the whole body according to the theory of traditional Chinese medicine (TCM), have been widely used in China for thousands of years. Herein, we investigated the value of tongue images and the tongue coating microbiome in the diagnosis of gastric cancer (GC). Methods: From May 2020 to January 2021, we simultaneously collected tongue images and tongue coating samples from 328 patients with GC (all newly diagnosed with GC) and 304 non-gastric cancer (NGC) participants in China, and 16 S rDNA was used to characterize the microbiome of the tongue coating samples. Then, artificial intelligence (AI) deep learning models were established to evaluate the value of tongue images and the tongue coating microbiome in the diagnosis of GC. Considering that tongue imaging is more convenient and economical as a diagnostic tool, we further conducted a prospective multicentre clinical study from May 2020 to March 2022 in China and recruited 937 patients with GC and 1911 participants with NGC from 10 centres across China to further evaluate the role of tongue images in the diagnosis of GC. Moreover, we verified this approach in another independent external validation cohort that included 294 patients with GC and 521 participants with NGC from 7 centres. This study is registered at ClinicalTrials.gov, NCT01090362. Findings: For the first time, we found that both tongue images and the tongue coating microbiome can be used as tools for the diagnosis of GC, and the area under the curve (AUC) value of the tongue image-based diagnostic model was 0.89. The AUC values of the tongue coating microbiome-based model reached 0.94 using genus data and 0.95 using species data. The results of the prospective multicentre clinical study showed that the AUC values of the three tongue image-based models for GCs reached 0.88-0.92 in the internal verification and 0.83-0.88 in the independent external verification, which were significantly superior to the combination of eight blood biomarkers. Interpretation: Our results suggest that tongue images can be used as a stable method for GC diagnosis and are significantly superior to conventional blood biomarkers. The three kinds of tongue image-based AI deep learning diagnostic models that we developed can be used to adequately distinguish patients with GC from participants with NGC, even early GC and precancerous lesions, such as atrophic gastritis (AG). Funding: The National Key R&D Program of China (2021YFA0910100), Program of Zhejiang Provincial TCM Sci-tech Plan (2018ZY006), Medical Science and Technology Project of Zhejiang Province (2022KY114, WKJ-ZJ-2104), Zhejiang Provincial Research Center for Upper Gastrointestinal Tract Cancer (JBZX-202006), Natural Science Foundation of Zhejiang Province (HDMY22H160008), Science and Technology Projects of Zhejiang Province (2019C03049), National Natural Science Foundation of China (82074245, 81973634, 82204828), and Chinese Postdoctoral Science Foundation (2022M713203).

Rhodium(III)-Catalyzed C-H/O2 Dual Activation and Macrocyclization: Synthesis and Evaluation of Pyrido[2,1-a]isoindole Grafted Macrocyclic Inhibitors for Influenza H1N1.

The development of environment-friendly, step economic couplings to generate structurally diverse macrocyclic compounds is highly desirable but poses a marked challenge. Inspired by the C-H oxidation mechanism of cytochromes P450, an unprecedented and practical RhIII -catalyzed acylmethylation macrocyclization via C-H/O2 dual activation has been developed by us. The process of macrocyclization is facilitated by a synergic coordination from pyridine and ester group. Interestingly, the reaction mode derives from a three-component coupling which differs from established olefination and alkylation paths. Density functional theory (DFT) calculations and control experiments revealed the mechanism of this unique C-H/O2 dual activation. The newly achieved acylmethylation macrocyclic products and their derivatives showed a potent anti-H1N1 bioactivity, which may provide an opportunity for the discovery of novel anti-H1N1 macrocyclic leading compounds.

An endoplasmic reticulum-associated degradation-related E2-E3 enzyme pair controls grain size and weight through the brassinosteroid signaling pathway in rice.

Grain size is an important agronomic trait, but our knowledge about grain size determination in crops is still limited. Endoplasmic reticulum (ER)-associated degradation (ERAD) is a special ubiquitin proteasome system that is involved in degrading misfolded or incompletely folded proteins in the ER. Here, we report that SMALL GRAIN 3 (SMG3) and DECREASED GRAIN SIZE 1 (DGS1), an ERAD-related E2-E3 enzyme pair, regulate grain size and weight through the brassinosteroid (BR) signaling pathway in rice (Oryza sativa). SMG3 encodes a homolog of Arabidopsis (Arabidopsis thaliana) UBIQUITIN CONJUGATING ENZYME 32, which is a conserved ERAD-associated E2 ubiquitin conjugating enzyme. SMG3 interacts with another grain size regulator, DGS1. Loss of function of SMG3 or DGS1 results in small grains, while overexpression of SMG3 or DGS1 leads to long grains. Further analyses showed that DGS1 is an active E3 ubiquitin ligase and colocates with SMG3 in the ER. SMG3 and DGS1 are involved in BR signaling. DGS1 ubiquitinates the BR receptor BRASSINOSTEROID INSENSITIVE 1 (BRI1) and affects its accumulation. Genetic analysis suggests that SMG3, DGS1, and BRI1 act together to regulate grain size and weight. In summary, our findings identify an ERAD-related E2-E3 pair that regulates grain size and weight, which gives insight into the function of ERAD in grain size control and BR signaling.

A Novel Image Encryption Algorithm Based on Improved Arnold Transform and Chaotic Pulse-Coupled Neural Network.

Information security has become a focal topic in the information and digital age. How to realize secure transmission and the secure storage of image data is a major research focus of information security. Aiming at this hot topic, in order to improve the security of image data transmission, this paper proposes an image encryption algorithm based on improved Arnold transform and a chaotic pulse-coupled neural network. Firstly, the oscillatory reset voltage is introduced into the uncoupled impulse neural network, which makes the uncoupled impulse neural network exhibit chaotic characteristics. The chaotic sequence is generated by multiple iterations of the chaotic pulse-coupled neural network, and then the image is pre-encrypted by XOR operation with the generated chaotic sequence. Secondly, using the improved Arnold transform, the pre-encrypted image is scrambled to further improve the scrambling degree and encryption effect of the pre-encrypted image so as to obtain the final ciphertext image. Finally, the security analysis and experimental simulation of the encrypted image are carried out. The results of quantitative evaluation show that the proposed algorithm has a better encryption effect than the partial encryption algorithm. The algorithm is highly sensitive to keys and plaintexts, has a large key space, and can effectively resist differential attacks and attacks such as noise and clipping.

Maize FERONIA-like receptor genes are involved in the response of multiple disease resistance in maize.

Receptor-like kinases (RLKs) are key modulators of diverse cellular processes such as development and sensing the extracellular environment. FERONIA, a member of the CrRLK1L subfamily, acts as a pleiotropic regulator of plant immune responses, but little is known about how maize FERONIA-like receptors (FLRs) function in responding to the major foliar diseases of maize such as northern corn leaf blight (NLB), northern corn leaf spot (NLS), anthracnose stalk rot (ASR), and southern corn leaf blight (SLB). Here, we identified three ZmFLR homologous proteins that showed cell membrane localization. Transient expression in Nicotiana benthamiana proved that ZmFLRs were capable of inducing cell death. To investigate the role of ZmFLRs in maize, we used virus-induced gene silencing to knock down expression of ZmFLR1/2 and ZmFLR3 resulting in reduced reactive oxygen species production induced by flg22 and chitin. The resistance of maize to NLB, NLS, ASR, and SLB was also reduced in the ZmFLRs knockdown maize plants. These results indicate that ZmFLRs are positively involved in broad-spectrum disease resistance in maize.

Plant Peroxisome-Targeting Effector MoPtep1 Is Required for the Virulence of Magnaporthe oryzae.

Rice blast caused by Magnaporthe oryzae is one of the most serious fungous diseases in rice. In the past decades, studies have reported that numerous M. oryzae effectors were secreted into plant cells to facilitate inoculation. Effectors target host proteins to assist the virulence of pathogens via the localization of specific organelles, such as the nucleus, endoplasmic reticulum, chloroplast, etc. However, studies on the pathogenesis of peroxisome-targeting effectors are still limited. In our previous study, we analyzed the subcellular localization of candidate effectors from M. oryzae using the agrobacterium-mediated transient expression system in tobacco and found that MoPtep1 (peroxisomes-targeted effector protein 1) localized in plant peroxisomes. Here, we proved that MoPtep1 was induced in the early stage of the M. oryzae infection and positively regulated the pathogenicity, while it did not affect the vegetative growth of mycelia. Subcellular localization results showed that MoPtep1 was localized in the plant peroxisomes with a signal peptide and a cupredoxin domain. Sequence analysis indicated that the homologous protein of MoPtep1 in plant-pathogenic fungi was evolutionarily conserved. Furthermore, MoPtep1 could suppress INF1-induced cell death in tobacco, and the targeting host proteins were identified using the Y2H system. Our results suggested that MoPtep1 is an important pathogenic effector in rice blast.

MicroRNA-activated hydrogel scaffold generated by 3D printing accelerates bone regeneration.

Bone defects remain a major threat to human health and bone tissue regeneration has become a prominent clinical demand worldwide. The combination of microRNA (miRNA) therapy with 3D printed scaffolds has always posed a challenge. It can mimic physiological bone healing processes, in which a biodegradable scaffold is gradually replaced by neo-tissue, and the sustained release of miRNA plays a vital role in creating an optimal osteogenic microenvironment, thus achieving promising bone repair outcomes. However, the balance between two key factors - scaffold degradation behavior and miRNA release profile - on osteogenesis and bone formation is still poorly understood. Herein, we construct a series of miRNA-activated hydrogel scaffolds (MAHSs) generated by 3D printing with different crosslinking degree to screened the interplay between scaffold degradation and miRNA release in the osteoinduction activity both in vitro and in vivo. Although MAHSs with a lower crosslinking degree (MAHS-0 and MAHS-0.25) released a higher amount of miR-29b in a sustained release profile, they degraded too fast to provide prolonged support for cell and tissue ingrowth. On the contrary, although the slow degradation of MAHSs with a higher crosslinking degree (MAHS-1 and MAHS-2.5) led to insufficient release of miR-29b, their adaptable degradation rate endowed them with more efficient osteoinductive behavior over the long term. MAHS-1 gave the most well-matched degradation rate and miR-29b release characteristics and was identified as the preferred MAHSs for accelerated bone regeneration. This study suggests that the bio-adaptable balance between scaffold degradation behavior and bioactive factors release profile plays a critical role in bone regeneration. These findings will provide a valuable reference about designing miRNAs as well as other bioactive molecules activated scaffold for tissue regeneration.

An Insect- and Rain-Proof Net Raises the Production and Quality of Chinese Bayberry by Preventing Damage From Insects and Altering Bacterial Communities.

Chinese bayberry (Myrica rubra) is a popular, nutrient- and antioxidant-rich fruit in Asia. However, it is susceptible to Drosophila during ripening, which disrupts production and causes economic loss. This study compared the effects of insecticides, insect-proof nets (IPNs), and insect- and rain-proof nets (IRPNs) on Chinese bayberry production and quality. Drosophila was absent in fruits from IPN- or IRPN-treated trees but only significantly reduced by insecticides. IPNs and IRPNs significantly increased fruit diameter, weight, edible rate and the Brix/acid ratio, and IRPNs had the strongest effect. Analysis of 16S rDNA showed that fruits collected from differently treated trees had unique bacterial communities. In IRPN fruits, Acetobacter and Gluconobacter were significantly decreased, reducing sugar consumption and disease; in addition, PICRUSt analysis predicted imputed functional profiles related to carbohydrate and nitrogen metabolism and mineral transport for fruit growth and development. This study proposed the use of IRPNs for improving Chinese bayberry production and quality.

Exogenous application of Mn significantly increased Cd accumulation in the Cd/Zn hyperaccumulator Sedum alfredii.

Sedum alfredii is a Cd/Zn hyperaccumulator native to China, which was collected from a mined area where Mn content in soil was extremely high, together with Zn and Cd content. We investigated the tolerance and accumulation ability of Mn and its possible association with Cd hyperaccumulation in this plant species by using MP-AES, SR-µ-XRF, and RT-PCR. The results showed that the hyperaccumulating ecotype (HE) S. alfredii exhibited high tolerance to Mn and accumulating around 10,000 and 12,000 mg kg-1 Mn in roots and shoots, respectively, without exhibiting toxicity under 5000 mg kg-1 Mn treatment for 4 weeks. Exposure to Cd significantly reduced plant uptake of Mn. In contrast, exogenous Mn application significantly improved root uptake and root-to-shoot translocation of Cd, resulting in the increased Cd accumulation in the shoots of HE S. alfredii. SR-µ-XRF analysis demonstrated that high Mn (20 µM) exposure resulted in higher intensities of Cd localized in both stem vascular bundles and cortex, as well as leaf mesophyll cells, than in those treated with low Mn levels (0.2 µM or 2.0 µM). RT-PCR analysis of several genes possibly involved in Mn/Cd transportation showed that expression of SaNramp3 in roots was significantly reduced under high Mn exposure. These results suggested a significant interaction between Cd and Mn in the HE S. alfredii plants, possibly through their competition for transporters and theoretically provided a strategy to improve the efficiency of Cd extraction from polluted soils by this plant species, after using appropriate nutrient management of Mn.

Environment-Specific Probiotic Supernatants Modify the Metabolic Activity and Survival of Streptococcus mutans in vitro.

A range of studies showed probiotics like Streptococcus oligofermentans and Limosilactobacillus reuteri to inhibit the cariogenic activity and survival of Streptococcus mutans, possibly via the production of substances like H2O2, reuterin, ammonia and organic acids. We aimed to assess the environment-specific mechanisms underlying this inhibition. We cultured L. reuteri and S. oligofermentans in various environments; minimal medium (MM), MM containing glucose (MM+Glu), glycerol (MM+Gly), lactic acid (MM+Lac), arginine (MM+Arg) and all four substances (MM+all) in vitro. Culture supernatants were obtained and metabolite concentrations (reuterin, ammonia, H2O2, lactate) measured. S. mutans was similarly cultivated in the above six different MM variation media, with glucose being additionally added to the MM+Gly, MM+Lac, and MM+Arg group, with (test groups) and without (control groups) the addition of the supernatants of the described probiotic cultures. Lactate production by S. mutans was measured and its survival (as colony-forming-units/mL) assessed. L. reuteri environment-specifically produced reuterin, H2O2, ammonia and lactate, as did S. oligofermentans. When cultured in S. oligofermentans supernatants, lactate production by S. mutans was significantly reduced (p < 0.01), especially in MM+Lac+Glu and MM+all, with no detectable lactate production at all (controls means ± SD: 4.46 ± 0.41 mM and 6.00 ± 0.29 mM, respectively, p < 0.001). A similar reduction in lactate production was found when S. mutans was cultured in L. reuteri supernatants (p < 0.05) for all groups except MM+Lac+Glu. Survival of S. mutans cultured in S. oligofermentans supernatants in MM+Lac+Glu and MM+all was significantly reduced by 0.6-log10 and 0.5-log10, respectively. Treatment with the supernatant of L. reuteri resulted in a reduction in the viability of S. mutans in MM+Gly+Glu and MM+all by 6.1-log10 and 7.1-log10, respectively. Probiotic effects on the metabolic activity and survival of S. mutans were environment-specific through different pathways.

Plasma Proteins as Biomarkers of Mortality After Total Body Irradiation in Mice.

During large-scale acute radiation exposure, rapidly distinguishing exposed individuals from nonexposed individuals is necessary. Identifying those exposed to high and potentially lethal radiation doses, and in need of immediate treatment, is especially important. To address this and find plasma biomarkers to assess ionizing radiation-induced mortality in the early stages, mice were administered a whole-body lethal dose of γ radiation, and radiation-induced damage was evaluated. Multiple blood biomarkers were screened using an antibody array, followed by validation using enzyme-linked immunoassay. The results revealed that irradiation (IR)-induced mortality in mice and caused body weight and blood platelet losses in deceased mice compared to surviving mice. The levels of certain proteins differed after IR between these 2 groups. Specific proteins in preirradiated mice were also found to potentiate radiosensitivity. Plasma levels of interleukin (IL)-22, urokinase, resistin, and IL-6 were associated with radiation-induced mortality in irradiated mice and may be useful as potential mortality predictors. Our results suggest that estimating the levels of certain plasma proteins is a promising alternative to conventional cytogenetic biodosimetry to accurately identify individuals exposed to high radiation doses and those at risk of death due to exposure. This strategy would facilitate the rapid triage of individuals requiring immediate and intensive medical treatment.

A Cd/Zn Co-hyperaccumulator and Pb accumulator, Sedum alfredii, is of high Cu tolerance.

High sensitivity towards Cu toxicity is problematic when using some hyperaccumulator plants for phytoremediation of soils with mixed contamination of Cu. Sedum alfredii, a Cd/Zn co-hyperaccumulator and Pb accumulator, is widely used for remediation of Cd, Zn, and Pb co-contaminated soils in China. In this paper, the tolerance and accumulation ability of S. alfredii towards Cu stress and its potential for phytoremediation of multi-metal polluted soils have been studied. Both the hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) of S. alfredii accumulated high Cu in the roots and translocated minimal Cu to the shoots, and Cu in the stems and leaves mostly restricted in the vascular tissues (phloem zone). The HE plants, however, exhibited high Cu resistance with stimulated lateral root growth and increased chlorophyll content under 10 µM Cu treatment. XANES analysis showed that Cu in HE roots comprised Cu2+ (46.7%), Cu-histidine (35.2%) and Cu-cell wall (18.1%). The NHE under Cu stress showed decreased biomass, reduced leaf chlorophyll content, altered root architecture, and higher Cu localized to root cell wall as compared with the HEs. Potted HE plants thrived six months in multi-metal contaminated soils including 3897 mg kg-1 available Cu. In conclusion, HE S alfredii is highly tolerant toward Cu due to metal homeostasis in root cells. Therefore, this plant has great potential to remediate Zn, Cd, and Pb contaminated soils those also contain high levels of Cu.

Introducing selective agrochemical manipulation of gibberellin metabolism into a cereal crop.

Use of growth retardants enables post-planting optimization of vegetative growth, which is particularly important given ongoing climate change. Mepiquat chloride is an economical and safe retardant widely applied in cotton farming, but it is not uniformly effective. Here, identification of its molecular target as the ent-copalyl diphosphate synthase that initiates gibberellin biosynthesis enabled the introduction of selective agrochemical inhibition, leaving intact more specialized metabolism important for resistance to biotic and abiotic stresses.

Effects of 3-dimensional Bioprinting Alginate/Gelatin Hydrogel Scaffold Extract on Proliferation and Differentiation of Human Dental Pulp Stem Cells.

INTRODUCTION: Alginate/gelatin hydrogel (Alg-Gel) scaffold has been applied in tissue engineering, but the research on its application in dental tissues regeneration is still lacking. We investigated the effect of this scaffold on human dental pulp stem cells (hDPSCs). METHODS: hDPSCs were cultured in both Alg-Gel and 3D-printed Alg-Gel scaffolds. Cell growth and adhesion were compared using fluorescein isothiocyanate-phalloidin staining and scanning electron microscopic micrographs. Changes in the proliferation in hDPSCs cultured in the complete culture medium containing aqueous extracts of the Alg-Gel or 3D-printed Alg-Gel scaffolds were examined using Cell Counting Kit-8 assay and flow cytometry analysis. Cells were cultured in the mineralization medium containing aqueous extracts of the Alg-Gel or 3D-printed Alg-Gel scaffolds for 7 or 14 days, and the differentiation of cells was shown by alizarin red S staining and alkaline phosphatase staining. The messenger RNA and protein expression of mineralization-related genes were detected with real-time polymerase chain reaction and Western blotting. Elemental analysis was used to test the material extract composition. RESULTS: More cells were grown and adhered to the 3D-printed Alg-Gel scaffolds than the Alg-Gel scaffolds. The aqueous extracts of 3D-printed scaffolds can promote cell proliferation, and compared with Alg-Gel scaffolds, the extracts of 3D-printed scaffolds were more effective. Compared with the negative control group, 3D-printed Alg-Gel scaffold and Alg-Gel scaffold aqueous extracts promoted osteogenic/odontoblastic differentiation of hDPSCs with the enhanced formation of bone-like nodules and the alkaline phosphatase staining. The expression of mineralization-related genes was also up-regulated. 3D-printed scaffold aqueous extract contained more calcium and phosphorus ions than the Alg-Gel scaffold. CONCLUSIONS: These findings suggest that compared with the Alg-Gel scaffold, 3D-printed Alg-Gel is more suitable for the growth of hDPSCs, and the scaffold extracts can better promote cell proliferation and differentiation.