Interesting Bioactivities Research Articles

Revealing synergistic mechanism of multiple components in Stauntonia brachyanthera Hand.-Mazz. for gout by virtual screening and system pharmacological approach

Stauntonia brachyanthera Hand.-Mazz. (SB), reported as a traditional Chinese medicine, displays a wide spectrum of interesting bioactivities, such as anti-inflammatory and analgesia. It is noteworthy that anti-gout effects of the components in SB have been reported. Hence, this study contributes to the prediction of promising active compounds and mechanisms for the treatment of gout. The active compounds with better oral bioavailability, and drug-likeness of SB were selected for further investigation by the approach of network pharmacology, molecular docking, gene ontology (GO) analysis, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis, respectively. A total of 34 predicted targets and 98 compounds in SB were obtained. Sorted by structure types of compounds, phenylethanoid glycosides exhibited the best anti-gout activity, followed by phenolics and flavonoids. What’s more, it was shown in the network analysis that Serine/threonine-protein kinase mTOR (mTOR), Mitogen-activated protein kinase 12 (MAPK12), tumor necrosis factor (TNF-α), Integrin alpha-4 (ITGA4) and Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit gamma (PIK3CG) were the key targets with intensely interaction, which should be attached more attention for further study. The functional enrichment analysis indicated that SB probably produced the anti-gout effects by synergistically regulating many biological pathways, such as MAPK signaling pathway, PI3K-Akt signaling pathway, Toll-like receptor signaling pathway and NOD-like receptor signaling pathway, etc. In addition, C61, C67, C68 and C81 might be promising leading compounds with good molecular docking score. As a consequence, the active constituents and mechanisms based on data analysis were holistically illuminated, which was of vital importance to the development of new drugs for gout.

Quantitative Analysis of Polysaccharide Composition in Polyporus umbellatus by HPLC-ESI-TOF-MS.

Polyporus umbellatus is a well-known and important medicinal fungus in Asia. Its polysaccharides possess interesting bioactivities such as antitumor, antioxidant, hepatoprotective and immunomodulatory effects. A qualitative and quantitative method has been established for the analysis of 12 monosaccharides comprising polysaccharides of Polyporus umbellatus based on high-performance liquid chromatography coupled with electrospray ionization–ion trap–time of flight–mass spectrometry. The hydrolysis conditions of the polysaccharides were optimized by orthogonal design. The results of optimized hydrolysis were as follows: neutral sugars and uronic acids 4 mol/L trifluoroacetic acid (TFA), 6 h, 120 °C; and amino sugars 3 mol/L TFA, 3 h, 100 °C. The resulting monosaccharides derivatized with 1-phenyl-3-methyl-5-pyrazolone have been well separated and analyzed by the established method. Identification of the monosaccharides was carried out by analyzing the mass spectral behaviors and chromatography characteristics of 1-phenyl-3-methyl-5-pyrazolone labeled monosaccharides. The results showed that polysaccharides in Polyporus umbellatus were composed of mannose, glucosamine, rhamnose, ribose, lyxose, erythrose, glucuronic acid, galacturonic acid, glucose, galactose, xylose, and fucose. Quantitative recoveries of these monosaccharides in the samples were in the range of 96.10–103.70%. This method is simple, accurate, and sensitive for the identification and quantification of monosaccharides, and can be applied to the quality control of Polyporusumbellatus as a natural medicine.

Ácidos hidroxicinámicos en producción animal: farmacocinética, farmacodinamia y sus efectos como promotor de crecimiento. Revisión

El uso de aditivos de origen natural en producción animal ha tomado gran importancia en el sector pecuario, debido a su potencial de promover el crecimiento de una forma similar a los compuestos sintéticos como hormonas y antibióticos, pero sin causar posibles daños a la salud del animal, del consumidor o detrimento en la calidad de la carne. En los aditivos de origen natural existe una amplia variedad de compuestos, que son extraídos de distintas partes de las plantas, donde se toman ciertos aceites esenciales, mezclas de compuestos o compuestos aislados para utilizarse como remedios medicinales o suplementos alimenticios. Dentro de estos extractos, se encuentran los ácidos hidroxicinámicos, presentes en una gran variedad de vegetales, frutas y granos; los cuales presentan interesantes propiedades bioactivas como son, antioxidantes, antimicrobianos, preventivos de enfermedades cardiovasculares e inmunomoduladores. El uso de este tipo de aditivos en producción animal aún es limitado, pero se sugiere que su inclusión puede ser favorable como una estrategia para promover el crecimiento; sin embargo, dos aspectos importantes a estudiarse en los ácidos hidroxicinámicos es su farmacocinética y farmacodinamia, y a partir de allí establecer las condiciones de dosis, períodos de uso y efectos, además las posibles rutas y biotransformaciones que pueden ocurrir en el organismo animal. Esta revisión discute sobre la inclusión de ácidos hidroxicinámicos en dietas de animales de engorda, propiedades farmacocinéticas y farmacodinamias, y los hallazgos como promotores de crecimiento y sus efectos en la calidad de la carne.

Development of a high performance thin layer chromatography method for the rapid qualification and quantification of phenolic compounds and abscisic acid in honeys

Honey is a natural product with a complex chemical composition consisting of sugars and other bioactive compounds. It is important in many traditional systems of medicine, exhibiting interesting bioactivities, in particular antimicrobial, anti-inflammatory and antioxidant effects. Authentication of botanical origin of honeys is particularly important in this context. Therefore, methods for quality control of honey and detection of its adulteration are very important.A HPTLC method for the quantitative determination of phenolic compounds in honey was developed for the first time. Seven phenolic compounds were detected and determined quantitatively in lime and acacia honey samples. The obtained results show that the HPTLC profiles of phenolic compounds and abscisic acid can be useful for determining the floral origin of honeys. Chromatographic tests were supplemented by statistical analysis (PCA and HCA) led to the successful separation of acacia and lime honey samples.

Synthesis and Acaricidal- and Insecticidal-Activity Evaluation of Novel Oxazolines Containing Sulfiliminyl Moieties and Their Derivatives.

Sulfimides and sulfoximines are highly relevant for medicinal chemistry and crop protection, as the resulting products can reveal interesting bioactivities. Herein, we report the design and synthesis of a series of novel 2,4-diphenyl-1,3-oxazolines containing sulfiliminyl and sulfoximinyl moieties. The acaricidal and insecticidal activities of the new compounds were evaluated and indicated that these compounds exhibited excellent acaricidal activities against spider mite larvae and eggs. The LC50 values of 6a-7, 6b-3, 6b-4, 6c-2, and 6c-4 against spider mite larvae were about 4 to 6 times lower than that of the commercial insecticide etoxazole (0.0221 mg L-1), and the LC50 value of 6a-4 against spider mite eggs was 0.0006 mg L-1, which was 10 times lower than that of etoxazole (0.0063 mg L-1). At the same time, most of the compounds showed insecticidal activity though their structure-activity relationships that were different. Oxazolines containing an N-cyano sulfiliminyl moiety at the para position of the 4-phenyl group exhibited better insecticidal activities against cotton bollworm and corn borer than etoxazole, whereas the compounds containing groups derived from sulfiliminyl and sulfoximinyl had weak insecticidal activities. This research again proved that the substituent type at the para site of the 4-phenyl moiety has a decisive role on the biological activity and insecticidal spectrum.

Assessment of multi-step processes for an integral use of the biomass of the marine microalga Amphidinium carterae

Sustainable dinoflagellate microalgae-based bioprocess designed to produce secondary metabolites (SMs) with interesting bioactivities are attracting increasing attention. However, dinoflagellates also produce other valuable bioproducts (e.g polyunsaturated fatty acids, carotenoids, etc.) that could be recovered and should therefore be taken into account in the bioprocess. In this study, biomass of the marine dinoflagellate microalga Amphidinium carterae was used to assess and optimise three different methods in order to obtain three families of high-value biochemical compounds present in the biomass. The existing processes encompassed a multi-step extraction process for carotenoids, fatty acids and APDs individually and are optimized for the integral valorization of raw A. carterae biomass, with SMs being the primary target compounds. Total process recovery yields were 97% for carotenoids, 80% for total fatty acids and 100% for an extract rich in APDs (not purified).

Computational prediction of the bioactivity potential of proteomes based on expert knowledge

Advances in the field of genome sequencing have enabled a comprehensive analysis and annotation of the dynamics of the protein inventory of cells. This has been proven particularly rewarding for microbial cells, for which the majority of proteins are already accessible to analysis through automatic metagenome annotation. The large-scale in silico screening of proteomes and metaproteomes is key to uncover bioactivities of translational, clinical and biotechnological interest, and to help assign functions to certain proteins, such as those predicted as hypothetical. This work introduces a new method for the prediction of the bioactivity potential of proteomes/metaproteomes, supporting the discovery of functionally relevant proteins based on prior knowledge. This methodology complements functional annotation enrichment methods by allowing the assignment of functions to proteins annotated as hypothetical/putative/uncharacterised, as well as and enabling the detection of specific bioactivities and the recovery of proteins from defined taxa. This work shows how the new method can be applied to screen proteome and metaproteome sets to obtain predictions of clinical or biotechnological interest based on reference datasets. Notably, with this methodology, the large information files obtained after DNA sequencing or protein identification experiments can be associated for translational purposes that, in cases such as antibiotic-resistance pathogens or foodborne diseases, may represent changes in how these important and global health burdens are approached in the clinical practice. Finally, the Sequence-based Expert-driven pRoteome bioactivity Prediction EnvironmENT, a public Web service implemented in Scala functional programming style, is introduced as means to ensure broad access to the method as well as to discuss main implementation issues, such as modularity, extensibility and interoperability.

Bioactive Secondary Metabolites from Octocoral-Associated Microbes-New Chances for Blue Growth.

Octocorals (Cnidaria, Anthozoa Octocorallia) are magnificent repositories of natural products with fascinating and unusual chemical structures and bioactivities of interest to medicine and biotechnology. However, mechanistic understanding of the contribution of microbial symbionts to the chemical diversity of octocorals is yet to be achieved. This review inventories the natural products so-far described for octocoral-derived bacteria and fungi, uncovering a true chemical arsenal of terpenes, steroids, alkaloids, and polyketides with antibacterial, antifungal, antiviral, antifouling, anticancer, anti-inflammatory, and antimalarial activities of enormous potential for blue growth. Genome mining of 15 bacterial associates (spanning 12 genera) cultivated from Eunicella spp. resulted in the identification of 440 putative and classifiable secondary metabolite biosynthetic gene clusters (BGCs), encompassing varied terpene-, polyketide-, bacteriocin-, and nonribosomal peptide-synthase BGCs. This points towards a widespread yet uncharted capacity of octocoral-associated bacteria to synthetize a broad range of natural products. However, to extend our knowledge and foster the near-future laboratory production of bioactive compounds from (cultivatable and currently uncultivatable) octocoral symbionts, optimal blending between targeted metagenomics, DNA recombinant technologies, improved symbiont cultivation, functional genomics, and analytical chemistry are required. Such a multidisciplinary undertaking is key to achieving a sustainable response to the urgent industrial demand for novel drugs and enzyme varieties.

Analysis of the Genome and Metabolome of Marine Myxobacteria Reveals High Potential for Biosynthesis of Novel Specialized Metabolites

Comparative genomic/metabolomic analysis is a powerful tool to disclose the potential of microbes for the biosynthesis of novel specialized metabolites. In the group of marine myxobacteria only a limited number of isolated species and sequenced genomes is so far available. However, the few compounds isolated thereof so far show interesting bioactivities and even novel chemical scaffolds; thereby indicating a huge potential for natural product discovery. In this study, all marine myxobacteria with accessible genome data (n = 5), including Haliangium ochraceum DSM 14365, Plesiocystis pacifica DSM 14875, Enhygromyxa salina DSM 15201 and the two newly sequenced species Enhygromyxa salina SWB005 and SWB007, were analyzed. All of these accessible genomes are large (~10 Mb), with a relatively small core genome and many unique coding sequences in each strain. Genome analysis revealed a high variety of biosynthetic gene clusters (BGCs) between the strains and several resistance models and essential core genes indicated the potential to biosynthesize antimicrobial molecules. Polyketides (PKs) and terpenes represented the majority of predicted specialized metabolite BGCs and contributed to the highest share between the strains. BGCs coding for non-ribosomal peptides (NRPs), PK/NRP hybrids and ribosomally synthesized and post-translationally modified peptides (RiPPs) were mostly strain specific. These results were in line with the metabolomic analysis, which revealed a high diversity of the chemical features between the strains. Only 6–11% of the metabolome was shared between all the investigated strains, which correlates to the small core genome of these bacteria (13–16% of each genome). In addition, the compound enhygrolide A, known from E. salina SWB005, was detected for the first time and structurally elucidated from Enhygromyxa salina SWB006. The here acquired data corroborate that these microorganisms represent a most promising source for the detection of novel specialized metabolites.

In vitro susceptibility of Microsporum spp. and mammalian cells to Eugenia caryophyllus essential oil, eugenol and semisynthetic derivatives.

Microsporum spp. are keratinophilic dermatophytes that mainly invade the stratum corneum of the skin and hair causing clinical symptoms associated with tinea. Its treatment has several limitations, and the search for new active molecules is necessary. To evaluate the antifungal and cytotoxic potential of Eugenia caryophyllus essential oil (EO), eugenol, isoeugenol and methylisoeugenol against Microsporum canis, M.gypseum and Vero cells. The EO was extracted by conventional heating-assisted hydrodistillation, the eugenol obtained commercially and the derivatives through Williamson synthesis. Minimal inhibitory concentration (MICs), minimum fungicidal concentration, inhibition of radial mycelial growth and germination inhibition were used to evaluate the antifungal activity. In addition, a colorimetric test was conducted to evaluate cytotoxic activity. MIC and MFC values for all compounds were 62.5-500μg/mL for both of the species of Microsporum evaluated. Also, concentrations of 300μg/mL of the compounds inhibited 100% of M.canis mycelium. The inhibition of germination was observed after 6hours of treatment (11.86±3.46-85.31±0%). No cytotoxicity was observed in Vero cells (CC50 >105μg/mL), whereas terbinafine showed CC50 31.00±0.61μg/mL. Our study indicates an interesting bioactivity of isoeugenol and methylisoeugenol against M.canis, M.gypseum and mammalian cells.

Antioxidant and angiotensin I-converting enzyme inhibitory activities of Xuanwei ham before and after cooking and in vitro simulated gastrointestinal digestion.

Xuanwei ham is especially rich in a large amount of peptides and free amino acids under the action of protein degradation. Some of these peptides can potentially exert bioactivities of interest for human health. Traditionally, Xuanwei ham should undergo Chinese household cooking treatments before eating. However, it has not been known how its bioactivity changes after cooking and gastrointestinal digestion. Herein, Xuanwei ham is analysed before and after cooking, as well as gastrointestinal digestion being simulated so as to evaluate and compare its effect on antioxidant and angiotensin I-converting enzyme (ACE) inhibitory activities. The antioxidant activity is analysed using five different methods, and results demonstrate that cooking has some negative effects on antioxidative capacity when determined using different antioxidant methods except for a significant increment in 1,1'-diphenyl-2-picrylhydrazyl radical-scavenging activity, while ACE inhibitory activity increases significantly after cooking compared with control samples. After gastrointestinal digestion of samples, there is a significant increment of the antioxidant and ACE inhibitory activities in comparison with control and cooked samples. Particularly, after gastrointestinal digestion, free thiols content and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical-cation-scavenging activity of Xuanwei ham, respectively, increase about twice and fourfold, while ACE inhibitory activity increases about twice compared to cooked samples, reaching the value of 83.73%. Therefore, through cooking the antioxidant activity and ACE inhibitory activity of Xuanwei ham are not completely lost and a part of them is still maintained, while gastrointestinal digestion produces a significant enhancement in both bioactivities, highlighting a greater potential for a beneficial physiological effect on human health after eating it.

Synthesis and Absolute Configuration of Natural 2‐Pyrones

2‐Pyrones are frequently produced by microorganisms and often exhibit interesting bioactivities. Therefore, a short and easy synthetic access to these natural products is desirable. Synthetic routes to nectriapyrone, gibepyrone A, racemic gulypyrone A, (+)‐germicidin C, (ent)‐desoxygermicidin C and (ent)‐prolipyrone A via a modular approach are presented, allowing the assignment of the absolute configurations of the latter three chiral compounds. The method failed for the synthesis of (ent)‐phomapyrone B that was thus synthesized via a different route, resulting in an assignment of the absolute configuration of natural phomapyrone B.

Mining for Microbial Gems: Integrating Proteomics in the Postgenomic Natural Product Discovery Pipeline.

Natural products (NPs) are a major source of compounds for medical, agricultural, and biotechnological industries. Many of these compounds are of microbial origin, and, in particular, from Actinobacteria or filamentous fungi. To successfully identify novel compounds that correlate to a bioactivity of interest, or discover new enzymes with desired functions, systematic multiomics approaches have been developed over the years. Bioinformatics tools harness the rapidly expanding wealth of genome sequence information, revealing previously unsuspected biosynthetic diversity. Varying growth conditions or application of elicitors are applied to activate cryptic biosynthetic gene clusters, and metabolomics provide detailed insights into the NPs they specify. Combining these technologies with proteomics‐based approaches to profile the biosynthetic enzymes provides scientists with insights into the full biosynthetic potential of microorganisms. The proteomics approaches include enrichment strategies such as employing activity‐based probes designed by chemical biology, as well as unbiased (quantitative) proteomics methods. In this review, the opportunities and challenges in microbial NP research are discussed, and, in particular, the application of proteomics to link biosynthetic enzymes to the molecules they produce, and vice versa.

Synthesis and production of the antitumor polyketide aurovertins and structurally related compounds.

Aurovertins belong to a family of highly reducing polyketides sharing a polyene α-pyrone-type structure. These compounds comprise aurovertin, asteltoxin, avertoxin, citreoviridin, verrucosidin, and their derivatives, which exihibit potent antitumor, antiviral, and antibacterial activities. Until now, over 40 aurovertins and structurally related compounds have been found in the fungal kingdom. Due to the unique structural feature and interesting bioactivities, significant progresses have been achieved for the structural identification, chemical synthesis, and biosynthesis of the mentioned compounds. Understanding of aurovertin biosynthetic mechanism provides a solid basis for engineering the metabolic pathway of those compounds by rational design and realizing their production in the model fungal host.

UPLC-ESI-QTOF-MS2 characterisation of Cola nitida resin fractions with inhibitory effects on NO and TNF-α released by LPS-activated J774 macrophage and on Trypanosoma cruzi and Leishmania amazonensis.

The resin of Cola nitida is used in western Cameroon as incense for spiritual protection and during ritual ceremonies. This plant secretion has never been investigated although previous chemical and biological studies on other resins have drawn many attentions. The resin fractions which revealed inhibitory effect on nitric oxide (NO) and tumour necrosis factor alpha (TNF-α) released by lipopolysaccharide (LPS)-activated J774 macrophage as well as on intracellular forms of Leishmania amazonensis and Trypanosoma cruzi amastigote were chemically characterised. Moreover, their antiparasitic activities were compared to those of semi-synthetic triterpenes. The anti-inflammatory activity was evaluated by measuring the nitrite production and the TNF-α concentration in the supernatants of LPS-activated macrophages by antigen capture enzyme-linked immunosorbent assay. Moreover, the antiparasitic assay was performed by infecting the host cells (THP-1) in a ratio parasite/cell 10:1 (L. amazonensis) and 2:1 (T. cruzi) and then exposed to the samples. The resin was separated in vacuo by liquid chromatography because of its sticky behaviour and the chemical profiles of the obtained fractions (F1-F4) were established by dereplication based on UPLC-ESI-MS2 data while semi-synthetic triterpenes were prepared from α-amyrin by oxidation reactions. Fractions F1-F4 inhibited NO and TNF-α almost similarly. However, only F1, F3 and F4 showed promising antiparasitic activities while F2 was moderately active against both parasites. Hence, F1-F4 were exclusively composed of pentacyclic triterpenes bearing oleanane and ursane skeletons. Semi-synthetic compounds revealed no to moderate antiparasitic activity compared to the fractions. Although it will be difficult to prove the interaction resin-spirit, interesting bioactivities were found in the resin fractions.

Chemical composition of barks from Quercus faginea trees and characterization of their lipophilic and polar extracts.

The bark from Quercus faginea mature trees from two sites was chemically characterized for the first time. The barks showed the following composition: ash 14.6%, total extractives 13.2%, suberin 2.9% and lignin 28.2%. The polysaccharides were composed mainly of glucose and xylose (50.3% and 35.1% of all monosaccharides respectively) with 4.8% of uronic acids. The suberin composition was: ω-hydroxyacids 46.3% of total compounds, ɑ,ω-alkanoic diacids 22.3%, alkanoic acids 5.9%, alkanols 6.7% and aromatics 6.9% (ferulic acid 4.0%). Polar extracts (ethanol-water) had a high phenolic content of 630.3 mg of gallic acid equivalents (GAE)/g of extract, condensed tannins 220.7 mg of catechin equivalents (CE)/g extract, and flavonoids 207.7 mg CE/g of extract. The antioxidant activity was very high corresponding to 1567 mg Trolox equivalents/g of extract, and an IC50 of 2.63 μg extract/ml. The lipophilic extracts were constituted mainly by glycerol and its derivatives (12.3% of all compounds), alkanoic acids (27.8%), sterols (11.5%) and triterpenes (17.8%). In view of an integrated valorization, Quercus faginea barks are interesting sources of polar compounds including phenols and polyphenols with possible interesting bioactivities, while the sterols and triterpenes contained in the lipophilic extracts are also valuable bioactive compounds or chemical intermediates for specific high-value market niches, such as cosmetics, pharmaceuticals and biomedicine.

Synthetic explorations of the briarane jungle: progress in developing a synthetic route to a common family of diterpenoid natural products.

The briarane diterpenoids are a large family of marine natural products that have been primarily isolated from gorgonian octocorals around the world. Structurally, the family is characterized by a trans-fused bicyclo[8.4.0]tetradecane ring system containing a central, stereogenic, all-carbon quaternary carbon (C1) flanked by three additional stereocentres (C2, C10, C14). Many family members have demonstrated biological activity in numerous areas, including: cytotoxicity, anti-inflammatory, antiviral, antifungal, immunomodulatory and insect control. Despite their interesting structural properties and bioactivity, the briaranes have been largely overlooked by the synthetic community. However, in recent years, several research groups have reported progress toward developing a synthetic route to these natural products. Most of these efforts have focused on the stereoselective construction of the central C1–C2–C10–C14 stereotetrad. This review will discuss the various synthetic efforts aimed at the briarane diterpenoids along with the challenges that remain.

Thioether-bridged arylalkyl-linked N-phenylpyrazole derivatives: Design, synthesis, insecticidal activities, structure-activity relationship and molecular-modeling studies

Owing to thioether diverse physicochemical properties by non-covalent interactions with bio-macromolecules, thioether derivatives containing heterocyclic moiety are known for their interesting insecticidal bioactivities and attracting considerable attention as neuroactive insecticides. Here we synthesis a series of novel thioether bridged N-phenylpyrazole derivatives incorporating various (hetero)aromatic substituents into 4-position of the pyrazole ring. Structure-activity relationship (SAR) studies resulted in compounds 6d and 7d with the most potent insecticidal activity among the series containing various substituted benzene substituents (LC50 = 13.70–25.47 μg/g). Further optimization to increase the lipophilicity and charge density of aromatic substituents of compounds 6d and 7d resulted in compounds 12d, 14d and 16d with sulfur-containing heterocycle substituents possessing good insecticidal activity against Musca domestica L. among the series (LC50 = 0.67–1.30 μg/g). The thioether bridge N-phenylpyrazole derivatives, which exhibit different length of the spacer arm introduced between N-phenylpyrazole moiety and the (hetero)aromatic substituents, were also prepared and evaluated. By contrast, the insecticidal activities of compounds containing the short thioether bridge, 1,2-bis((hetero)aromatic thio) ethane, are higher than that containing the long thioether bridge, 1,3-bis((hetero)aromatic thio) propane. The results of molecular docking and pharmacophore analyses indicated A299, T303, and L306 of a subunit were essential to form non-covalent interactions contacts with the ligands. Specially, the sulfur-containing heterocycle substituent derivatives 12d and 14d as the sterically favored areas could form the important hydrophobic interactions with the deeper residue P295.

Cytotoxic Effect of Aeruginosin-865, Resveratrol and Capsaicin on Mouse Fibroblasts and Cells Derived from Fallow Deer

Natural substances offer interesting bioactivity patterns including antiproliferative, antioxidant or cytotoxic effects. However, the safety profile of many of them has not been extensively determined. In this study, the cytotoxic effect of Aeruginosin-865, resveratrol and capsaicin at different concentrations was tested on normal mouse cells (NIH/3T3) and tumour fibroblasts (WEHI-13VAR) as well as on liver- and kidney-derived cells from fallow deer. A lactate dehydrogenase cytotoxicity assay kit was used to measure cell death in response to treatment with the test substances. It was found that NIH/3T3 cells tolerated Aeruginosin-865 (10-200 μM) and resveratrol (5-100 μM) treatment without any cytotoxic effect, while capsaicin exerted a cytotoxic effect only at the highest tested concentration (200 μ M). Mouse fibrosarcoma cells were more sensitive to the cytotoxic effect of all three compounds where Aeruginosin-865 (100-200 μM) and resveratrol (50–100 μM) showed high-dose cytotoxicity and capsaicin showed low- and high-dose cytotoxicity (25 μM and 200 μ M). The three tested compounds at the highest concentrations were found to be cytotoxic to both liver- and kidney-derived cells from fallow deer. Overall, the results indicate that the cytotoxic effects of the three tested natural substances on cells derived from fallow deer and mouse tumour fibroblasts differ significantly from those exerted on normal fibroblasts. The results demonstrate the potential of these natural compounds as therapeutic agents and pave the way for future in vivo toxicological investigations.

Anti-Phytopathogenic and Cytotoxic Activities of Crude Extracts and Secondary Metabolites of Marine-Derived Fungi.

Thirty-one isolates belonging to eight genera in seven orders were identified from 141 strains that were isolated from several marine plants. Alternaria sp. and Fusarium sp. were found to be the predominant fungi. Evaluation of the anti-phytopathogenic bacterial and fungal activities, as well as the cytotoxicity of these 31 extracts, revealed that most of them displayed different levels of bioactivities. Due to their interesting bioactivities, two fungal strains—Fusarium equiseti (P18) and Alternaria sp. (P8)—were selected for chemical investigation and compounds 1–4 were obtained. The structure of 1 was elucidated by 1D and 2D NMR analysis, as well as high-resolution electrospray ionization mass spectroscopy (HRESIMS), and the absolute configuration of its stereogenic carbon (C-11) was established by comparison of the experimental and calculated electronic circular-dichroism (ECD) spectra. Moreover, alterperylenol (4) exhibited antibacterial activity against Clavibacter michiganensis with a minimum inhibitory concentration (MIC) of 1.95 μg/mL, which was 2-fold stronger than that of streptomycin sulfate. Additionally, an antibacterial mechanism study revealed that 4 caused membrane hyperpolarization without evidence of destruction of cell membrane integrity. Furthermore, stemphyperylenol (3) displayed potent antifungal activity against Pestallozzia theae and Alternaria brassicicola with MIC values equal to those of carbendazim. The cytotoxicity of 1 and 2 against human lung carcinoma (A-549), human cervical carcinoma (HeLa), and human hepatoma (HepG2) cell lines were also evaluated.