Strong antagonism of an endophyte of Fraxinus excelsior towards the ash dieback pathogen, Hymenoscyphus fraxineus, is mediated by the antifungal secondary metabolite PF1140.

Ash dieback, caused by the fungal pathogen Hymenoscyphus fraxineus (Helotiales, Ascomycota), is threatening the existence of the European ash, Fraxineus excelsior. During our search for biological control agents for this devastating disease, endophytic fungi were isolated from healthy plant tissues and co-cultivated with H. fraxineus to assess their antagonistic potential. Among the strains screened, Penicillium cf. manginii DSM 104493 most strongly inhibited the pathogen. Initially, DSM 104493 showed promise in planta as a biocontrol agent. Inoculation of DSM 104493 into axenically cultured ash seedlings greatly decreased the development of disease symptoms in seedlings infected with H. fraxineus. The fungus was thus cultivated on a larger scale in order to obtain sufficient material to identify active metabolites that accounted for the antibiosis observed in dual culture. We isolated PF1140 (1) and identified it as the main active compound in the course of a bioassay-guided isolation strategy. Furthermore, its derivative 2, the mycotoxin citreoviridin (3), three tetramic acids of the vancouverone type (4-6), and penidiamide (7) were isolated by preparative chromatography. The structures were elucidated mainly by NMR spectroscopy and high-resolution mass spectrometry (HRMS), of which compounds 2 and 6 represent novel natural products. Of the compounds tested, not only PF1140 (1) strongly inhibited H. fraxineus in an agar diffusion assay but also showed phytotoxic effects in a leaf puncture assay. Unfortunately, both the latent virulent attributes of DSM 104493 observed subsequent to these experiments in planta and the production of mycotoxins exclude strain Penicillium cf. manginii DSM 104493 from further development as a safe biocontrol agent.IMPORTANCEEnvironmentally friendly measures are urgently needed to control the causative agent of ash dieback, Hymenoscyphus fraxineus. Herein, we show that the endophyte DSM 104493 exhibits protective effects in vitro and in planta. We traced the activity of DSM 104493 to the antifungal natural product PF1140, which unfortunately also showed phytotoxic effects. Our results have important implications for understanding plant-fungal interactions mediated by secondary metabolites, not only in the context of ash dieback but also generally in plant-microbial interactions.

Sensitivity to NH3 Vapor: Synthesis and Characterization of Five New Coordination Polymers Based on 2,2-Dimethylglutaric Acid and Bis(triazole)-Derived Ligands.

Five new coordination polymers (CPs) were obtained as a result of hydrothermal reactions of 2,2-dimethylglutaric acid (H2dmg) and 1,4-bis(1H-1,2,4-triazol-1-ylmethyl)benzene (pbtx)/1,2-bis(1H-1,2,4-triazol-1-ylmethyl)benzene (obtx) ligands with some metal ions [Co(µ-dmg)(µ-obtx)]n (1), [Zn(µ-dmg)(µ-obtx)]n (2), [Cd(µ-dmg)(µ-obtx)]n (3), [Co2(µ-dmg)2(µ-pbtx)2]n (4), and [Cd(µ-dmg)(H2O)(µ-pbtx)]n (5). All of the compounds were characterized by elemental analysis, FT-IR spectroscopy, single-crystal X-ray diffraction, powder X-ray diffraction, and thermal analysis techniques. The single-crystal X-ray studies show that all compounds exhibit 2D layer structures. To examine the ammonia sensing properties of five new coordination complexes (1-5), the absorption and emission spectra of CPs embedded in ethyl cellulose thin films were measured by exposure to different concentrations of ammonia (NH3) vapor. The [Co2(µ-dmg)2(µ-pbtx)2]n (4)-based sensor agent was found to show promising sensor properties in detecting NH3 vapor.

Hepatopulmonary Shunt Ratio Verification Model for Transarterial Radioembolization.

INTRODUCTION: The most important toxicity of transarterial radioembolization therapy applied in liver malignancies is radiation pneumonitis and fibrosis due to hepatopulmonary shunt of Yttrium-90 (90Y) microspheres. Currently, Technetium-99m macroaggregated albumin (99mTc-MAA) scintigraphic images are used to estimate lung shunt fraction (LSF) before treatment. The aim of this study was to create a phantom to calculate exact LFS rates according to 99mTc activities in the phantom and to compare these rates with LSF values calculated from scintigraphic images. MATERIALS AND METHODS: A 3D-printed lung and liver phantom containing two liver tumors was developed from Polylactic Acid (PLA) material, which is similar to the normal-sized human body in terms of texture and density. Actual %LSFs were calculated by filling phantoms and tumors with 99mTc radionuclide. After the phantoms were placed in the water tank made of plexiglass material, planar, SPECT, and SPECT/CT images were obtained. The actual LSF ratio calculated from the activity amounts filled into the phantom was used for the verification of the quantification of scintigraphic images and the results obtained by the Simplicit 90YTM method. RESULTS: In our experimental model, LSFs calculated from 99mTc activities filled into the lungs, normal liver, small tumor, and large tumor were found to be 0%, 6.2%, 10.8%, and 16.9%. According to these actual LSF values, LSF values were calculated from planar, SPECT/CT (without attenuation correction), and SPECT/CT (with both attenuation and scatter correction) scintigraphic images of the phantom. In each scintigraphy, doses were calculated for lung, small tumor, large tumor, normal liver, and Simplicit 90YTM. The doses calculated from planar and SPECT/CT (NoAC+NoSC) images were found to be higher than the actual doses. The doses calculated from SPECT/CT (with AC+with SC) images and Simplicit 90YTM were found to be closer to the real dose values. CONCLUSION: LSF is critical in dosimetry calculations of 90Y microsphere therapy. The newly introduced hepatopulmonary shunt phantom in this study is suitable for LSF verification for all models/brands of SPECT and SPECT/CT devices.

Bioactive Compounds from an Endophytic Pezicula sp. Showing Antagonistic Effects against the Ash Dieback Pathogen.

A fungal endophyte originating from the Canary Islands was identified as a potent antagonist against the fungal phytopathogen Hymenoscyphus fraxineus, which causes the devastating ash dieback disease. This endophyte was tentatively identified as Pezicula cf. ericae, using molecular barcoding. Isolation of secondary metabolites by preparative high-performance liquid chromatography (HPLC) yielded the known compounds CJ-17,572 (1), mycorrhizin A (3) and cryptosporioptides A-C (4-6), besides a new N-acetylated dihydroxyphenylalanin derivative 2, named peziculastatin. Planar structures were elucidated by NMR and HRMS data, while the relative stereochemistry of 2 was assigned by H,H and C,H coupling constants. The assignment of the unknown stereochemistry of CJ-17,572 (1) was hampered by the broadening of NMR signals. Nevertheless, after semisynthetic conversion of 1 into its methyl derivatives 7 and 8, presumably preventing tautomeric effects, the relative configuration could be assigned, whereas comparison of ECD data to those of related compounds determined the absolute configuration. Metabolites 1 and 3 showed significant antifungal effects in vitro against H. fraxineus. Furthermore, 4-6 exhibited significant dispersive effects on preformed biofilms of S. aureus at concentrations up to 2 µg/mL, while the biofilm formation of C. albicans was also inhibited. Thus, cryptosporioptides might constitute a potential source for the development of novel antibiofilm agents.

Air-Assisted Alkanol-Based Nanostructured Supramolecular Liquid-Liquid Microextraction for Extraction and Spectrophotometric Determination of Morin in Fruit and Beverage Samples.

A new air-assisted liquid-liquid microextraction method based on alkanol nanostructured supramolecular solvents coupled to spectrometric analysis was developed for extraction, preconcentration, and spectrophotometric determination of morin. Al(III)-morin complex was performed at pH 4.5. Four different alkanol-based SUPRAS (supramolecular solvents) were prepared for the separation and preconcentration of Al-morin complex from aqueous solution by using vortex and centrifugation. Effect of analytical variables and tolerance limit of matrix ions were investigated. Under the optimum conditions, detection limit, quantification limit, relative standard deviation, preconcentration factor, and enhancement factor were found as 3.5 µg L-1, 10 µg L-1, 3.1%, 120, and 95, respectively. The accuracy of the method was performed with standard addition. The obtained results demonstrated the applicability of the method for the separation, preconcentration, and determination of morin in fruit and beverage samples. The method also complies with green chemistry principles as it uses green solvents, reduces reagent volumes, and produces low amounts of waste. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12161-021-02111-3.

A new magnetic nanodiamond/graphene oxide hybrid (Fe3O4@ND@GO) material for pre-concentration and sensitive determination of sildenafil in alleged herbal aphrodisiacs by HPLC-DAD system.

A sensitive analytical methodology was investigated to concentrate and determine of sildenafil citrate (SLC) present at trace level in herbal supplementary products. The proposed method is based on simple and sensitive pre-concentration of SLC by using magnetic solid phase extraction with new developed magnetic nanodiamond/graphene oxide hybrid (Fe3O4@ND@GO) material as a sorbent. Experimental variables affecting the extraction efficiency of SLC like; pH, sample volume, eluent type and volume, extraction time and amount of adsorbent were studied and optimized in detail. Determination of sildenafil citrate after magnetic solid phase extraction (MSPE) was carried out by HPLC-DAD system. The morphology, composition, and properties of the synthesized hybrid material was characterized by Fourier transform infrared spectrometry (FT-IR), Raman spectrometry (Raman), X-ray diffraction spectrometry (XRD), scanning electron microscopy (SEM), mapping photographs, zeta potential analyzer, and BET surface area analysis. Under optimized conditions, linear range was ranged from 5.00 to 250.00 ng mL-1 with R2 of 0.9952. The limit of detection (LOD) was 1.49 ng mL-1 and the recoveries at two spiked levels were ranged from 94.0 to 104.1% with the relative standard deviation (RSD) < 7.1% (n = 5). The enhancement factor (EF) was 86.9. The results show that the combination MSPE with HPLC-DAD is a suitable and sensitive method for the determination of SLC in real samples.