Ultrasonic-assisted extraction of total flavonoids from Zanthoxylum bungeanum residue and their allelopathic mechanism on Microcystis aeruginosa.

Water eutrophication has emerged as a pressing concern for massive algal blooms, and these harmful blooms can potentially generate harmful toxins, which can detrimentally impact the aquatic environment and human health. Consequently, it is imperative to identify a safe and efficient approach to combat algal blooms to safeguard the ecological safety of water. This study aimed to investigate the procedure for extracting total flavonoids from Z. bungeanum residue and assess its antioxidant properties. The most favorable parameters for extracting total flavonoids from Z. bungeanum residue were a liquid-solid ratio (LSR) of 20 mL/g, a solvent concentration of 60%, an extraction period of 55 min, and an ultrasonic temperature of 80 °C. Meanwhile, the photosynthetic inhibitory mechanism of Z. bungeanum residue extracts against M. aeruginosa was assessed with a particular focus on the concentration-dependent toxicity effect. Z. bungeanum residue extracts damaged the oxygen-evolving complex structure, influenced energy capture and distribution, and inhibited the electron transport of PSII in M. aeruginosa. Furthermore, the enhanced capacity for ROS detoxification enables treated cells to sustain their photosynthetic activity. The findings of this study hold considerable relevance for the ecological management community and offer potential avenues for the practical utilization of resources in controlling algal blooms.

Cyanobacteria-Mediated Light-Driven Biotransformation: The Current Status and Perspectives.

Most chemicals are manufactured by traditional chemical processes but at the expense of toxic catalyst use, high energy consumption, and waste generation. Biotransformation is a green, sustainable, and cost-effective process. As cyanobacteria can use light as the energy source to power the synthesis of NADPH and ATP, using cyanobacteria as the chassis organisms to design and develop light-driven biotransformation platforms for chemical synthesis has been gaining attention, since it can provide a theoretical and practical basis for the sustainable and green production of chemicals. Meanwhile, metabolic engineering and genome editing techniques have tremendous prospects for further engineering and optimizing chassis cells to achieve efficient light-driven systems for synthesizing various chemicals. Here, we display the potential of cyanobacteria as a promising light-driven biotransformation platform for the efficient synthesis of green chemicals and current achievements of light-driven biotransformation processes in wild-type or genetically modified cyanobacteria. Meanwhile, future perspectives of one-pot enzymatic cascade biotransformation from biobased materials in cyanobacteria have been proposed, which could provide additional research insights for green biotransformation and accelerate the advancement of biomanufacturing industries.

Insights into water freezing from classical nucleation theory.

Water freezing is a crucial physical phenomenon. The process of ice formation, and the estimation of the ice nucleation rate also have important applications. However, until now, the experimental phenomenon of rapid freezing of water in nanoseconds has not been fully explained theoretically, and the physics underlying the experimental phenomenon has still not been revealed. In this work, combining classical nucleation theory with Mie theory, a kinetic model is developed that reproduces for the first time the experimental phenomenon of decreasing transmissivity. The process of ice formation (nucleation, growth and engulfment) has been revealed. In the process of theoretical derivation, the Zel'dovich-Frenkel (ZF) equation is developed, indicating a limit to the phase transition driving force |Δµ|/(kBT) ≤ 1. By analyzing the experimental and simulation results, it is suggested that the change in the transparency of the sample may be caused by the ice/vacuum interface scattering. In addition, during the rapid phase transition, it was found that the phase transition continues to occur even after phase fraction normalization. Finally, the approximate formula between the nucleation rate and sample transparency is given. This formula can predict the change of sample transparency during phase transition and provides a way to measure the nucleation rate. The results presented here give an insight into the phase transition kinetics, and the methodology may also work for the phase transitions of other materials.

Molecular mechanism of GylR-mediated regulation of glycerol metabolism in Streptomyces clavuligerus NRRL 3585.

Glycerol is a readily available and low-cost simple polyol compound, which can be used as a carbon source for microorganisms to produce various value-added products. Understanding the underlying regulatory mechanism in glycerol metabolism is critical for making better use of glycerol for diverse applications. In a few reported Streptomyces strains, the glycerol utilization gene cluster (glp operon) was shown to be regulated by the IclR family transcriptional regulator GylR. However, the molecular regulatory mechanism mediated by GylR has not been fully elucidated. In this study, we first analyzed the available Actinobacteria genomes in the NCBI Genome database, and found that the glp operon-like gene clusters are conserved in Streptomyces and several other genera of Actinobacteria. By taking Streptomyces clavuligerus NRRL 3585 as a model system, we identified that GylR represses the expressions of glp operon and gylR by directly binding to their promoter regions. Both glycerol-3-phosphate and dihydroxyacetone phosphate can induce the dissociation of GylR from its binding sequences. Furthermore, we identified a minimal essential operator site (a palindromic 18-bp sequence) of GylR-like regulators in Streptomyces. Our study for the first time reported the binding sequences and effector molecules of GylR-like proteins in Streptomyces. The molecular regulatory mechanism mediated by GylR presumably exists widely in Streptomyces. Our findings would facilitate the design of glycerol utilization pathways for producing valuable products. Moreover, our study provided new basic elements for the development of glycerol-inducible regulatory tools for synthetic biology research in the future.

Net Learning.

Graph neural networks, which generalize deep learning to graph-structured data, have achieved significant improvements in numerous graph-related tasks. Petri nets (PNs), on the other hand, are mainly used for the modeling and analysis of various event-driven systems from the perspective of prior knowledge, mechanisms, and tasks. Compared with graph data, net data can simulate the dynamic behavioral features of systems and are more suitable for representing real-world problems. However, the problem of large-scale data analysis has been puzzling the PN field for decades, and thus, limited its universal applicability. In this article, a framework of net learning (NL) is proposed. NL contains the advantages of PN modeling and analysis with the advantages of graph learning computation. Then, two kinds of NL algorithms are designed for performance analysis of stochastic PNs, and more specifically, the hidden feature information of the PN is obtained by mapping net information to the low-dimensional feature space. Experiments demonstrate the effectiveness of the proposed model and algorithms on the performance analysis of stochastic PNs.

The Role of Root Morphology and Pulling Direction in Pullout Resistance of Alfalfa Roots.

There is a growing consensus on soil conservation by mechanics of plant root system. In order to further study how root system exerts its mechanical properties during soil reinforcing process and which morphological indicator is suitable for reflecting pullout resistance, in-situ vertical pullout test (VPT) and 45° oblique pullout test (OPT) were performed on alfalfa (Medicago sativa L.) roots in the loess area. The results showed that the failure mode of alfalfa roots was pulling out in this study. The peak pullout resistance of the roots increased with root diameter, root length and root surface area, and power law relationships were observed between the pullout resistance and the morphological indices: root diameter, root length and root surface area. The maximum gray relational degree of the morphological indices was 0.841 (VPT) and 0.849 (OPT) for root surface area, suggesting that root surface area was a more significant root morphological index affecting root pullout resistance than root diameter and root length, and was more suitable for characterizing the difference in peak pullout resistance of roots with different size. The index could be used to validate the methods for predicting root pullout capacity. The value of peak pullout resistance was 17.2 ± 2.3 N in VPT test and 28.2 ± 3.8 N (mean ± SE) in OPT test, and a significant difference was observed between the two tests, which showed that the pulling direction significantly affected the peak pullout resistance of alfalfa roots. Vertical pullout test, giving the safety margin, was suggested to determine root pullout resistance for estimate of root reinforcement.

An Optical Crack Growth Sensor Using the Digital Sampling Moiré Method.

High-accuracy crack growth measurement is crucial for the health assessment of concrete structures. In this work, an optical crack growth sensor using the digital sampling moiré (DSM) method is developed for two-dimensional (2D) crack growth monitoring. The DSM method generates moiré fringes from a single image through digital image processing, and it measures 2D displacements using the phase difference of moiré fringes between motion. Compared with the previous sensors using traditional photogrammetric algorithms such as the normalized cross-correlation (NCC) method, this new DSM-based sensor has several advantages: First, it is of a higher sensitivity and lower computational cost; second, it requires no prior calibration to get accurate 2D displacements which can greatly simplify the practical application for multiple crack monitoring. In addition, it is more robust to the change of imaging distance, which is determined by the height difference between two sides of a concrete crack. These advantages break the limitation of the NCC method and broaden the applicability of the crack growth sensor. These advantages have been verified with one numerical simulation and two laboratory tests.

Insights into grapevine defense response against drought as revealed by biochemical, physiological and RNA-Seq analysis.

Grapevine is an important and extensively grown fruit crop, which is severely hampered by drought worldwide. So, comprehending the impact of drought on grapevine genetic resources is necessary. In the present study, RNA-sequencing was executed using cDNA libraries constructed from both drought-stress and control plants. Results generated 12,451 differentially expressed genes (DEGs), out of which 8,021 genes were up-regulated, and 4,430 were down-regulated. Further physiological and biochemical investigations were also performed to validate the biological processes associated with the development of grapevine in response to drought stress. Results also revealed that decline in the rate of stomatal conductance, in turn, decrease the photosynthetic activity and CO2 assimilation in the grapevine leaves. Reactive oxygen species, including stress enzymes and their related proteins, and secondary metabolites were also activated in the present study. Likewise, various hormones also induced in response to drought stress. Overall, the present study concludes that these DEGs play both positive and negative roles in drought tolerance by regulating various biological pathways of grapevine. Nevertheless, our findings have provided valuable gene information for future studies of abiotic stress in grapevine and various other fruit crops.

Enzyme activities and gene expression of starch metabolism provide insights into grape berry development.

Grapes are categorized as a non-climacteric type of fruit which its ripening is not associated to important rises in respiration and ethylene synthesis. The starch metabolism shares a certain role in the carbohydrate metabolic pathways during grape berry development, and is regarded as an important transient pool in the pathway of sugar accumulation. However, the comprehensive role of starch and its contribution to the quality and flavor of grape berry have not been explored thoroughly. In this study, the expression levels of genes enzyme activities and carbohydrate concentrations related to starch metabolism, were analyzed to understand the molecular mechanism of starch accumulation during grape berry development. The results indicated that starch granules in grape berry were located at the chloroplast in the sub-epidermal tissues, acting as the temporary reserves of photosynthetic products to meet the needs for berry development, and relatively high starch contents could be detected at véraison stage. Moreover, both ADP-glucose pyrophosphorylase (EC 2.7.7.27) and sucrose phosphate synthase (EC 2.3.1.14) involved in starch synthesis displayed elevated gene expression and enzymes activities in the sub-epidermal tissue, while α- and ß-amylases involved in its degradation were highly transcribed and active in the central flesh, explaining the absence of starch in this last tissue. Change in the gene expression and activities of ADP-glucose pyrophosphorylase, ß-amylase and sucrose phosphate synthase revealed that they were regulated by the circadian rhythms in the fruitlets compared with those in the leaves. Both the morphological, enzymological and transcriptional data in this study provide advanced understandings on the function of starch during berry development and ripening that are so important for berry quality. This study will further facilitate our understanding of the sugar metabolism in grape berry as well as in other plant species.

Synthesis and cytotoxic activity of novel tetrahydrobenzodifuran-imidazolium salt derivatives.

The synthesis of a series of novel 4-substituted 2,3,6,7-tetrahydrobenzo [1,2-b;4,5-b']difuran-1H-imidazolium salts is presented. The biological properties of the compounds were evaluated in vitro against a panel of human tumor cell lines. Results suggest that the 5,6-dimethyl-benzimidazole or 2-methyl-benzimidazole ring, and substitution of the imidazolyl-3-position with a 2-naphthylmethyl substituent or 2-naphthylacyl substituent, were important to the cytotoxic activity. Notably, 3-(2-Naphthylmethyl)-1-((2,3,6,7-tetrahydrobenzo[1,2-b;4,5-b']difuran-4-yl)methyl)-1H-5,6-dimethyl-benzimidazol-3-ium bromide (42) was found to be the most potent derivative against five human tumor cell lines with IC50 values of 1.06-4.34µM and more selective towards SMMC-7721, A549 and SW480 cell lines. 3-(2-Naphthylacyl)-1-((2,3,6,7-tetrahydrobenzo[1,2-b;4,5-b']difuran-4-yl)methyl)-1H-2-methyl-benzimidazol-3-ium bromide (37) showed higher selectivity to SMMC-7721 and MCF-7 cell lines with IC50 values 2.7-fold and 8.4-fold lower than DDP. Study regarding to the antitumor mechanism of action showed that compound 37 could induce cell cycle G1 phase arrest and apoptosis in SMMC-7721 cells.

Characterization of miR061 and its target genes in grapevine responding to exogenous gibberellic acid.

MicroRNAs (miRNAs), as an important growth regulator, are also involved in gibberellic acid (GA) signaling, revealing much relationship between miRNAs and GA in various plant responses. Grape is highly sensitive to GA3, which plays a significant regulatory role in regulation of flower development, berry expansion, berry set, berry ripening, and seedlessness induction; further, it was found that grapevine miR061 (VvmiR061) is a GA3 responsive miRNA. In this study, grapevine REV (VvREV) and HOX32 (VvHOX32), two target genes of VvmiR061, were predicted, verified, and cloned; homologous conservation was analyzed in various plants. The expression profiles of both VvmiR061 and its target genes (VvREV and VvHOX32) under GA3 treatment were detected by qRT-PCR during grapevine flower and berry development. Results revealed that GA3 treatment has upregulated the transcription of VvREV and VvHOX32, while it downregulated the expression of VvmiR061. The function of VvmiR061 in cleaving target genes VvREV and VvHOX32 was diminished by GA3 treatment during flower developmental process. The results of this study exhibited the importance of VvmiR061 in regulating flower development and GA3 signaling pathway and also contributed some to the knowledge of small RNA-mediated regulation in grape.

Structure and Function of a C-C Bond Cleaving Oxygenase in Atypical Angucycline Biosynthesis.

C-C bond ring cleaving oxygenases represent a unique family of enzymes involved in the B ring cleavage reaction only observed in atypical angucycline biosynthesis. B ring cleavage is the key reaction leading to dramatic divergence in the final structures of atypical angucyclines. Here, we present the crystal structure of AlpJ, the first structure of this family of enzymes. AlpJ has been verified as the enzyme catalyzing C-C bond cleavage in kinamycin biosynthesis. The crystal structure of the AlpJ monomer resembles the dimeric structure of ferredoxin-like proteins. The N- and C-terminal halves of AlpJ are homologous, and both contain a putative hydrophobic substrate binding pocket in the "closed" and "open" conformations, respectively. Structural comparison of AlpJ with ActVA-Orf6 and protein-ligand docking analysis suggest that the residues including Asn60, Trp64, and Trp181 are possibly involved in substrate recognition. Site-directed mutagenesis results supported our hypothesis, as mutation of these residues led to nearly a complete loss of the activity of AlpJ. Structural analysis also revealed that AlpJ possesses an intramolecular domain-domain interface, where the residues His50 and Tyr178 form a hydrogen bond that probably stabilizes the three-dimensional structure of AlpJ. Site-directed mutagenesis showed that the two residues, His50 and Tyr178, were vital for the activity of AlpJ. Our findings shed light on the structure and catalytic mechanism of the AlpJ family of oxygenases, which presumably involves two active sites that might function in a cooperative manner.

Synthesis and cytotoxic activity of novel hexahydropyrrolo[2,3-b]indole imidazolium salts.

A series of novel hexahydropyrrolo[2,3-b]indole-1H-imidazolium salts were synthesized and evaluated in vitro against a panel of human tumor cell lines. The results suggest that the 5,6-dimethyl-benzimidazole ring, and substitution of the imidazolyl-3-position with a 2-bromobenzyl or 2-naphthylmethyl group, were important for the cytotoxic activity. Notably, Compound 43, bearing a 2-bromobenzyl substituent at position-3 of 5,6-dimethyl-benzimidazole, was found to possess the most potent derivative against five human tumor cell lines with IC50 values below 2.68µM and more selective towards SMMC-7721, A549 and SW480 cell lines. Compounds 25 and 39 were more selective to HL-60 and MCF-7 cell lines with IC50 values of 0.47 and 1.46µM.

[Pharmacodynamic comparison between sanhuang decoction for purging stomach-fire and its concentrated granule].

OBJECTIVE: To discuss the feasibility that a traditional decoction is substituted by its concentrated gratnule. METHODS: The effects of the two kinds of decoction were compared on mice auricle tumefaction induced by xylene, arresting bleeding of mice broken tails and mice alvine creepage. RESULT: There was no remarkable difference between two kinds of decoction on pharmacodynamic effects except purging action. CONCLUSION: The research and exploitation of classical prescription concentrated granule has great signifaction.

[Calculation of spectroscopic properties of Tm:YVO4 crystals].

Sigma and pi polarized absorption spectra of Tm:YVO4 crystal have been measured at room temperature. Considering the difference of the sigma and pi polarized absorption spectra and the changing of the refractive index with different wavelengths, we calculate the intensity parameters of Tm3+ in crystal YVO4 which are omega 2 = 1.9416 x 10(-20) (cm2), omega 4 = 0.1568 x 10(-20) (cm2), omega 6 = 0.3963 x 10(-20) (cm2) by Judd-Ofelt theory. The spectra characteristic parameters, such as radiative transition rates, luminescence branching ratio, total radiative lifetime and integral cross-section are also calculated. The results show that the spontaneous rate of the transition 1D2-->3F4 is much higher than that of the transition 1D2 to other levels.