Design, Synthesis, and Antifungal Evaluation of Diverse Heterocyclic Hydrazide Derivatives as Potential Succinate Dehydrogenase Inhibitors.

Plant pathogenic fungi pose a significant threat to agricultural production, necessitating the development of new and more effective fungicides. The ring replacement strategy has emerged as a highly successful approach in molecular design. In this study, we employed the ring replacement strategy to successfully design and synthesize 32 novel hydrazide derivatives containing diverse heterocycles, such as thiazole, isoxazole, pyrazole, thiadiazole, 1,3,4-oxadiazole, 1,2,4-oxadiazole, thiophene, pyridine, and pyrazine. Their antifungal activities were evaluated in vitro and in vivo. Bioassay results revealed that most of the title compounds displayed remarkable antifungal activities in vitro against four tested phytopathogenic fungi, including Fusarium graminearum, Botrytis cinerea, Sclerotinia sclerotiorum, and Rhizoctonia solani. Especially, compound 5aa displayed a broad spectrum of antifungal activity against F. graminearum, B. cinerea, S. sclerotiorum, and R. solani, with the corresponding EC50 values of 0.12, 4.48, 0.33, and 0.15 µg/mL, respectively. In the antifungal growth assay, compound 5aa displayed a protection efficacy of 75.5% against Fusarium head blight (FHB) at a concentration of 200 µg/mL. In another in vivo antifungal activity evaluation, compound 5aa exhibited a noteworthy protective efficacy of 92.0% against rape Sclerotinia rot (RSR) at a concentration of 100 µg/mL, which was comparable to the positive control tebuconazole (97.5%). The existing results suggest that compound 5aa has a broad-spectrum antifungal activity. Electron microscopy observations showed that compound 5aa might cause mycelial abnormalities and organelle damage in F. graminearum. Moreover, in the in vitro enzyme assay, we found that the target compounds 5aa, 5ab, and 5ca displayed significant inhibitory effects toward succinate dehydrogenase, with the corresponding IC50 values of 1.62, 1.74, and 1.96 µM, respectively, which were superior to that of boscalid (IC50 = 2.38 µM). Additionally, molecular docking and molecular dynamics simulation results revealed that compounds 5aa, 5ab, and 5ca have the capacity to bind in the active pocket of succinate dehydrogenase (SDH), establishing hydrogen-bonding interactions with neighboring amino acid residues.

Targeting Magnaporthe oryzae effector MoErs1 and host papain-like protease OsRD21 interaction to combat rice blast.

Effector proteins secreted by plant pathogenic fungi are important artilleries against host immunity, but there is no precedent of such effectors being explored as antifungal targets. Here we demonstrate that MoErs1, a species-specific effector protein secreted by the rice blast fungus Magnaporthe oryzae, inhibits the function of rice papain-like cysteine protease OsRD21 involved in rice immunity. Disrupting MoErs1-OsRD21 interaction effectively controls rice blast. In addition, we show that FY21001, a structure-function-based designer compound, specifically binds to and inhibits MoErs1 function. FY21001 significantly and effectively controls rice blast in field tests. Our study revealed a novel concept of targeting pathogen-specific effector proteins to prevent and manage crop diseases.

Design and Synthesis of Novel Diphenyl Ether Carboxamide Derivatives To Control the Phytopathogenic Fungus Sclerotinia sclerotiorum.

Sclerotinia stem rot (SSR) caused by the phytopathogenic fungus Sclerotinia sclerotiorum has led to serious losses in the yields of oilseed rape and other crops every year. Here, we designed and synthesized a series of carboxamide derivatives containing a diphenyl ether skeleton by adopting the scaffold splicing strategy. From the results of the mycelium growth inhibition experiment, inhibition rates of compounds 4j and 4i showed more than 80% to control S. sclerotiorum at a dose of 50 µg/mL, which is close to that of the positive control (flubeneteram, 95%). Then, the results of a structure-activity relationship study showed that the benzyl scaffold was very important for antifungal activity and that introducing a halogen atom on the benzyl ring would improve antifungal activity. Furthermore, the results of an in vitro activity test suggested that these novel compounds can inhibit the activity of succinate dehydrogenase (SDH), and the binding mode of 4j with SDH was basically similar to that of the flutolanil derivative. Morphological observation of mycelium revealed that compound 4j could cause a damage on the mycelial morphology and cell structure of S. sclerotiorum, resulting in inhibition of the growth of mycelia. Furthermore, in vivo antifungal activity assessment of 4j displayed a good control of S. sclerotiorum (>97%) with a result similar to that of the positive control at a concentration of 200 mg/L. Thus, the diphenyl ether carboxamide skeleton is a new starting point for the discovery of new SDH inhibitors and is worthy of further development.

Development and characterization of anti-IL-5 monoclonal antibody Fab fragment for blocking IL-5/IL-5Rα binding.

Interleukin-5 (IL-5) is a homodimeric cytokine that is a crucial regulator of the proliferation, activation, and maturation of eosinophils. Anti-IL-5 monoclonal antibodies, which block the binding of IL-5 to the IL-5 receptor subunit alpha (IL-5Rα), have been successfully used to treat eosinophilic (EOS) asthma. The currently marketed monoclonal antibody drugs require repeated injections for administration, which seriously affect patient compliance and high systemic exposure for injectable drug delivery. Here we successfully screened and developed the Fab (fragment of antigen binding), which is 1/3rd the molecular weight of IgG, favoring inhalation-mediated delivery to the lungs, making it more effective for asthma treatment. The 20A12-Fab-H12L3 can bind to IL-5 with a binding constant of 1.236E-09 M while significantly inhibiting the IL-5/IL-5Rα complex formation. We found that the light chain amino acids (S46 and F71) significantly affected the antibody expression during humanization. The 20A12-Fab-H12L3 significantly inhibited the proliferation of TF-1 cells and blocked the IL-5 binding to the IL-5Rα-overexpressing human embryonic kidney (HEK)-293 cells in vitro. Therefore, based on the mutant IL-5 binding with Fab, we explained why antibodies blocked IL-5 binding to IL-5Rα. Thus, this study provided a candidate pharmaceutical antibody for inhalation drug delivery.

Rapid and Sensitive Detection of Polycyclic Aromatic Hydrocarbons in Tea Leaves Using Magnetic Approach.

A rapid and efficient method using an alkyl-functionalized magnetic nanoparticles-based extraction technique combined with Ultra-High Performance Liquid Chromatography was developed for the detection of trace amounts of polycyclic aromatic hydrocarbons in tea leaves. As a popular coating for chromatographic column packing materials, C18-alkyl has been demonstrated to be effective in separating polycyclic aromatic hydrocarbons. Additionally, the magnetism of the nanomaterials accelerates the extraction process while their high surface ratio enables desirable dispersity in the sample matrix. Meanwhile, the adsorbents can be washed and reused 30 times without compromising recovery, which greatly reduces the budget. The effects of various parameters were investigated and optimized, and the recoveries for five analytes were in the range of 84.8-105.4%. The RSD of intra-day and inter-day were below 11.9% and 6.8%, respectively. The limits of detection and limits of quantification ranged from 1.69-9.97 ng g-1 and 5.12-30.21 ng g-1, indicating satisfactory sensitivity. Thus, the proposed methodology is rapid, highly efficient, and economical, and it expands the application of magnetic cleanup approaches in complex food matrices.

Association of working hours and cumulative fatigue among Chinese primary health care professionals.

Introduction: The association between long working hours and cumulative fatigue is widely acknowledged in the literature. However, there are few studies on the mediating effect of working hours on cumulative fatigue using occupational stress as a mediating variable. The present study aimed at investigating the mediating role of occupational stress in the relationship between working hours and cumulative fatigue in a sample of 1,327 primary health care professionals. Methods: The Core Occupational Stress Scale and the Workers' Fatigue Accumulation Self-Diagnosis Scale were utilized in this study. The mediating effect of occupational stress was examined using hierarchical regression analysis and the Bootstrap test. Results: Working hours were positively associated with cumulative fatigue via occupational stress (p < 0.01). Occupational stress was found to partially mediate the relationship between working hours and cumulative fatigue, with a mediating effect of 0.078 (95% CI: 0.043-0.115, p < 0.01), and the percentage of occupational stress mediating effect was 28.3%. Discussion: Working hours can be associated with cumulative fatigue either directly or indirectly via occupational stress. As a result, by reducing occupational stress, primary health care professionals may reduce the cumulative fatigue symptoms caused by long hours of work.

Potential of natural flavonols and flavanones in the treatment of ulcerative colitis.

Ulcerative colitis (UC) is an inflammatory bowel disease generally characterized by chronic, persistent, recurrent, and non-specific ulcers of the intestine. Its main clinical manifestations include abdominal pain, diarrhea, and bloody stools. This disease is difficult to cure and even carries the risk of canceration. It has been listed as a modern refractory disease by the World Health Organization. Though a large amount of drugs are available for the inhibition of UC, the conventional treatment such as aminosalicylic acids, glucocorticoids, immunosuppressors, and biological agents possess certain limitations and serious side effects. Therefore, it is urgently needed for safe and effective drugs of UC, and natural-derived flavonols and flavanones showed tremendous potential. The present study concentrated on the progress of natural-derived flavonols and flavanones from edible and pharmaceutical plants for the remedy of UC over the last two decades. The potential pharmaceutical of natural-derived flavonols and flavanones against UC were closely connected with the modulation of gut microflora, gut barrier function, inflammatory reactions, oxidative stress, and apoptosis. The excellent efficacy and safety of natural flavonols and flavanones make them prospective drug candidates for UC suppression.

Genome Mining and Heterologous Expression Guided the Discovery of Antimicrobial Naphthocyclinones from Streptomyces eurocidicus CGMCC 4.1086.

A type II polyketide synthase biosynthetic gene cluster (nap) was identified in Streptomyces eurocidicus CGMCC 4.1086 via genome mining. The heterologous expression of the cryptic nap gene cluster in Streptomyces albus J1074 generated dimerized aromatic polyketide naphthocyclinones (1-3), whose structures were determined via extensive analysis using nuclear magnetic resonance and high-resolution electrospray ionization mass spectroscopy. The biological pathway of naphthocyclinone synthesis was revealed via in vivo gene deletion, in vitro biochemical reactions, and comparative genomics. Remarkably, 3 played a crucial role in inhibiting Phytophthora capsici and Phytophthora sojae, with EC50 values of 6.1 and 20.2 µg/mL, respectively. Furthermore, 3 exhibited a potent protective effect against P. capsici and P. sojae in greenhouse tests.

Chemical Investigation of Endophytic Diaporthe unshiuensis YSP3 Reveals New Antibacterial and Cytotoxic Agents.

Chemical investigation of the plant-derived endophytic fungus Diaporthe unshiuensis YSP3 led to the isolation of four new compounds (1-4), including two new xanthones (phomopthane A and B, 1 and 2), one new alternariol methyl ether derivative (3) and one α-pyrone derivative (phomopyrone B, 4), together with eight known compounds (5-12). The structures of new compounds were interpreted on the basis of spectroscopic data and single-crystal X-ray diffraction analysis. All new compounds were assessed for their antimicrobial and cytotoxic potential. Compound 1 showed cytotoxic activity against HeLa and MCF-7 cells with IC50 values of 5.92 µM and 7.50 µM, respectively, while compound 3 has an antibacterial effect on Bacillus subtilis (MIC value 16 µg/mL).

New metabolites from Streptomyces pseudovenezuelae NA07424 and their potential activity of inducing resistance in plants against Phytophthora capsici.

BACKGROUND: The lack of novel fungicide and appearance of resistance are the most emergent problems in the control of Phytophthora diseases. Plant immunity elicitors that induce systemic resistance in plants are regarded as the new strategy for plant disease control. Streptomyces can produce a variety of bioactive natural products, which are important resources for lead compounds of plant immunity elicitors. RESULTS: A novel peptidendrocin C (1) together with the known analog peptidendrocin B (2) were isolated from Streptomyces pseudovenezuelae NA07424. Their structures were confirmed by spectroscopic data and Marfey's reaction. In bioactive assays, compound 1 played an important role in inducing systemic resistance of Nicotiana benthamiana against Phytophthora capsici growth, with a 90.5% inhibition ratio at 400 µg/mL, while compound 2 showed moderate activity, inhibiting P. capsici growth by a 50.8% decrease at 400 µg/mL. Simultaneously, two compounds promoted enhanced expression of the PR1 gene and callose accumulation in N. benthamiana and Arabidopsis thaliana. In this paper, we also provide the first insights into their biosynthesis by confirming their biosynthesis gene cluster and related functional genes. CONCLUSION: Our findings show that 1 and 2 have the potential to be used as lead compounds for development of new plant immunity elicitors to control Phytophthora diseases. The study of the biosynthesis pathway lays the groundwork for further application of the bioactive natural products. © 2022 Society of Chemical Industry.

Design, Synthesis, and Antifungal Activity of N-(alkoxy)-Diphenyl Ether Carboxamide Derivates as Novel Succinate Dehydrogenase Inhibitors.

Succinate dehydrogenase (SDH, EC 1.3.5.1) is one of the most promising targets for fungicide development and has attracted great attention worldwide. However, existing commercial fungicides targeting SDH have led to the increasingly prominent problem of pathogen resistance, so it is necessary to develop new fungicides. Herein, we used a structure-based molecular design strategy to design and synthesize a series of novel SDHI fungicides containing an N-(alkoxy)diphenyl ether carboxamide skeleton. The mycelial growth inhibition experiment showed that compound M15 exhibited a very good control effect against four plant pathogens, with inhibition rates of more than 60% at a dose of 50 µg/mL. A structure-activity relationship study found that N-O-benzyl-substituted derivatives showed better antifungal activity than others, especially the introduction of a halogen on the benzyl. Furthermore, the molecular docking results suggested that π-π interactions with Trp35 and hydrogen bonds with Tyr33 and Trp173 were crucial interaction sites when inhibitors bound to SDH. Morphological observation of mycelium revealed that M15 could inhibit the growth of mycelia. Moreover, in vivo and in vitro tests showed that M15 not only inhibited the enzyme activity of SDH but also effectively protected rice from damage due to R. solani infection, with a result close to that of the control at a concentration of 200 µg/mL. Thus, the N-(alkoxy)diphenyl ether carboxamide skeleton is a new starting point for the discovery of new SDH inhibitors and is worthy of further investigation.

Two new sesquiterpene derivatives, dendocarbin B and bisaborosaol C with antifungal activity from the endophytic fungus Nigrospora chinensis GGY-3.

Two novel sesquiterpene derivatives, dendocarbin B (1), bisaborosaol C (2), and nine known compounds (3-11), were isolated from Nigrospora chinensis GGY-3 derived from Ilex cornuta. The structures of new compounds were elucidated using HR-ESI-MS, 1 D and 2 D NMR spectra, X-ray diffraction analysis as well as ECD calculation and comparison. Compound 1 showed moderate antifungal activities against Rhizoctonia solani and Botrytis cinerea. Compounds 5 and 6 exhibited significant inhibitory activity against Phytophthora capsici, Magnaporthe oryzae and R. solani with EC50 values ranging from 13.91 to 29.49 µg/mL. Compounds 10 and 11 displayed moderate antibacterial effects on Bacillus subtilis and Xanthomonas oryzae pv. oryzae (Xoo), with MIC values of 16-64 µg/mL. Particularly, 11 presented strong antibacterial activity against Staphylococcus aureus with an MIC value of 4 µg/mL (2 µg/mL for streptomycin sulfate). In addition, compound 11 also possessed DPPH radical scavenging capability with an IC50 value of 14.80 µg/mL.

Potential of natural products in the treatment of irritable bowel syndrome.

BACKGROUND: Irritable bowel syndrome (IBS) is a kind of functional bowel disease that is characterized by bellyache, abdominal distension, and diarrhea. Although not life-threatening, IBS has a long course and recurrent attacks and seriously affects the life quality of patients. Current drugs for treating IBS possess remarkable limitations, such as limited efficacy and severe adverse reactions. Therefore, developing novel medications to treat IBS is quite essential, and natural products may be a substantial source. PURPOSE: This is the first systematic review elaborating the recent advancement of natural products as potential drugs for the therapy of IBS. METHODS: A comprehensive retrieval of studies was carried out in scientific databases including PubMed, Web of Science, Elsevier, and CNKI. By using ("irritable bowel syndrome" OR "IBS") AND ("natural product" OR "natural compound" OR "phytochemical") as keywords, the eligible studies were screened, and the relevant information about therapeutic action and mechanism of natural products treating IBS was extracted. RESULTS: Natural products against IBS consisted of four categories, namely, terpenoids, flavonoids, alkaloids, and phenols. Furthermore, the underlying mechanisms for natural products treating IBS were tightly associated with increased TJs and mucus protein expression, regulation of the brain-gut axis and gut microbiota structure, and inhibition of inflammatory response and intestinal mucosal damage. CONCLUSION: Natural products could be extremely prospective candidate drugs used to treat IBS, and further preclinical and clinical researches are needed to guarantee their efficacy and safety.

Discovery of Tropolone Stipitaldehyde as a Potential Agent for Controlling Phytophthora Blight and Its Action Mechanism Research.

The fermentation of endophytic Nigrospora chinensis GGY-3 resulted in the isolation of tropolone stipitaldehyde (1), which exhibited broad-spectrum inhibition activity against fungi and bacteria, especially against Phytophthora capsici, with an EC50 value of 0.83 µg/mL and Xanthomonas oryzae pv. oryzicola, with a minimum inhibitory concentration value of 4.0 µg/mL. The in vitro and in vivo assays demonstrated that 1 had a significant protective effect on P. capsici. Furthermore, 1 inhibited the spore germination of P. capsici and damaged the plasma membrane structure. As observed by SEM and TEM, after exposure to 1, mycelia exhibited swelling, shrunken, branch-increasing phenomena, cell wall and membrane damage, and disordered content. Transcriptome analysis revealed that 1 might affect starch and sucrose metabolism and fatty acid biosynthesis by suppressing the expression of genes relevant to cell wall synthetases and cell membrane-associated genes. These findings indicate that 1 may be a potential agrochemical fungicide for controlling phytophthora blight.

(+)- and (-)-trichodermatrione A: a pair of enantiomers with a cyclobutane-containing skeleton from the endophytic fungus Trichoderma sp. EFT2.

(±)-trichodermatrione A, a pair of cyclobutane-containing enantiomers with an undescribed tricyclic 6/4/6 skeleton, was isolated from Trichoderma sp. EFT2, an endophytic fungus from Euonymus fortunei (Turcz.) Hand.-Mazz (Celastraceae). The racemates were separated by chiral HPLC with the structures elucidated by a combination of MS, NMR, ECD calculation and X-ray crystallography analyses. (±)- trichodermatrione A and enantiomers were found to be antibacterial against phytopathogenic bacteria Xanthomonas oryzae pv. oryzae (Xoo) and X. oryzae pv. oryzicola (Xoc).

The Big Five Inventory-2 in China: A Comprehensive Psychometric Evaluation in Four Diverse Samples.

The Big Five Inventory-2 (BFI-2) has received wide recognition since its publication because it strikes a good balance between content coverage and brevity. The current study translated the BFI-2 into Chinese, evaluated its psychometric properties in four diverse Chinese samples (college students, adult employees, adults treated for substance use, and adolescents), and compared its factor structure with those obtained from two U.S. samples. Across two studies, the Chinese BFI-2 demonstrated good reliability (Cronbach's α and test-retest reliability), structural validity, convergent/discriminant validity, and criterion-related validity at the domain level. At lower levels of analyses, some facets and negatively worded items functioned better among participants with higher than those with lower education levels. Implications, limitations, and future directions are discussed.

Azo-incorporating Increases Inhibitory Activity of Succinate Dehydrogenase.

Diversity of pesticide discovery provided a solution to resistance. Here, we presented a strategy of azo-incorporating to promote the diverse developments of fungicide. A series of novel fungicides were synthesized by incorporating azobenzene derivatives into fluxapyroxad. Much better in vitro fungicidal activity increases for compound 9d were observed compared to the positive control, fluxapyroxad against Botrytis cinerea and Rhizoctonia solani. Compound 9d (IC50 = 0.03 µM) also had a great enzyme-inhibiting activity increase toward succinate dehydrogenase in comparison with fluxapyroxad (IC50 = 4.40 µM). A comparatively equivalent biological activity was observed between compounds 8a and 9d. SEM analysis helped us to observe clearly the morphology of the fungi before and after active ingredient delivery. Our results of molecular docking analysis, fluorescence quenching analysis, and enzymatic assays demonstrated that compound 8a and 9d act on SDH. An increase in inhibitory activity could be occurring after incorporation of azobenzene, which provided a new strategy for molecular design in pesticide discovery.

An Activity-Based Sensing Fluorogenic Probe for Monitoring Ethylene in Living Cells and Plants.

Ethylene (ET) is an important gaseous plant hormone. It is highly desirable to develop fluorescent probes for monitoring ethylene in living cells. We report an efficient RhIII -catalysed coupling of N-phenoxyacetamides to ethylene in the presence of an alcohol. The newly discovered coupling reaction exhibited a wide scope of N-phenoxyacetamides and excellent regioselectivity. We successfully developed three fluorophore-tagged RhIII -based fluorogenic coumarin-ethylene probes (CEPs) using this strategy for the selective and quantitative detection of ethylene. CEP-1 exhibited the highest sensitivity with a limit of detection of ethylene at 52 ppb in air. Furthermore, CEP-1 was successfully applied for imaging in living CHO-K1 cells and for monitoring endogenous-induced changes in ethylene biosynthesis in tobacco and Arabidopsis thaliana plants. These results indicate that CEP-1 has great potential to illuminate the spatiotemporal regulation of ethylene biosynthesis and ethylene signal transduction in living biological systems.

Eremophilane Sesquiterpenes and Benzene Derivatives from the Endophyte Microdiplodia sp. WGHS5.

Three new eremophilane sesquiterpenes phomadecalins G-I (1-3) and two new benzene derivatives microdiplzenes A and B (12 and 13), together with nine known eremophilane sesquiterpenes (4-11 and 14) were isolated from an endophytic fungus, Microdiplodia sp. WGHS5. Their structures were elucidated by the interpretation of HR-ESI-MS and NMR data; meanwhile, the absolute configurations of new compounds were determined on the base of ECD calculations. All compounds were evaluated for the antimicrobial activities and antiproliferative effect on human gastric cancer cell lines (BGC-823).

Antifungal Activity of Chaetoviridin A from Chaetomium globosum CEF-082 Metabolites Against Verticillium dahliae in Cotton.

Cotton Verticillium wilt (CVW) is a severe soilborne disease caused by the pathogen Verticillium dahliae, and it has a great impact on cotton production. Previous studies found that the biocontrol agent Chaetomium globosum CEF-082 and its metabolic filtrate could reduce the incidence of CVW; however, the underlying mechanism remains unclear. The metabolic crude extract of CEF-082 increased the sensitivity of V. dahliae to stress, degraded the cell wall of V. dahliae, and increased the emergence and plant height of cotton. Through separation and purification of the metabolic crude extract of CEF-082, chaetoviridin A was identified and found to be highly active against V. dahliae. The compound caused cell necrosis and mycelial deformation, increased the production of reactive oxygen species and nitrous oxide, and inhibited the germination of microsclerotia of V. dahliae, enhancing the cotton plant defense response. In addition, CEF-082 also colonized cotton plants.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.