Active Secondary Metabolites Research Articles

Endophyte Bacillus vallismortis BL01 to Control Fungal and Bacterial Phytopathogens of Tomato (Solanum lycopersicum L.) Plants

Some strains of Bacillus vallismortis have been reported to be efficient biocontrol agents against tomato pathogens. The aim of our study was to assess the biocontrol ability of the endophytic strain BL01 Bacillus vallismortis through in vitro and field trials, as well as to verify its plant colonization ability and analyze the bacterial genome in order to find genes responsible for the biocontrol activity. We demonstrated in a gnotobiotic system and by confocal laser microscopy that the endophytic strain BL01 was able to colonize the endosphere and rhizosphere of tomato, winter wheat and oilseed rape. In vitro experiments demonstrated the inhibition activity of BL01 against a wide range of phytopathogenic fungi and bacteria. BL01 showed biological efficacy in two-year field experiments with tomato plants against black bacterial spotting by 40–70.8% and against late blight by 47.1% and increased tomato harvest by 24.9% or 10.9 tons per hectare compared to the control. Genome analysis revealed the presence of genes that are responsible for the synthesis of biologically active secondary metabolites, which could be responsible for the biocontrol action. Strain BL01 B. vallismortis can be considered an effective biocontrol agent to control both fungal and bacterial diseases in tomato plants.

Antioxidant and anticancer activity of active secondary metabolites produced by Streptomyces griseorubens

Antioxidant and anticancer activity of active secondary metabolites produced by Streptomyces griseorubens

ENDOPHYTIC FUNGI AS CHITIN-MODIFYING ENZYMES PRODUCERS

Chitosan and chitooligosaccharides, which are products of chitin modification, have wide industrial applications. An improved understanding of the structure and properties of chitin-modifying enzymes has increased the interest in obtaining them from microbial sources and using them in the biotechnological production of these essential sugars. Endophytic fungi include a diverse group of microorganisms that inhabit the intercellular and intracellular areas of plant tissues, thus exerting beneficial effects on host species. They are considered an extremely valuable source of biologically active secondary metabolites and enzymes with high application potential. This review presents the potential of endophytic fungi to produce chitin-modifying enzymes and discusses the role of chitin modification by enzymes produced by fungal endophytes in their survival in the plant host.

A Review of Sponge-Derived Diterpenes: 2009-2022.

Sponges are a vital source of pharmaceutically active secondary metabolites, of which the main structural types are alkaloids and terpenoids. Many of these compounds exhibit biological activities. Focusing specifically on diterpenoids, this article reviews the structures and biological activities of 228 diterpenes isolated from more than 33 genera of sponges from 2009 to 2022. The Spongia sponges produce the most diterpenoid molecules among all genera, accounting for 27%. Of the 228 molecules, 110 exhibit cytotoxic, antibacterial, antifungal, antiparasitic, anti-inflammatory, and antifouling activities, among others. The most prevalent activity is cytotoxicity, present in 54 molecules, which represent 24% of the diterpenes reported. These structurally and biologically diverse diterpenoids highlight the vast, yet largely untapped, potential of marine sponges in the discovery of new bioactive molecules for medicinal use.

Silver and Iron Nanoparticles: Green Synthesis and Characterization.

Biosynthesized nanoparticles (NPs) are gaining traction because they leverage physiologically active plant secondary metabolites, enabling eco-friendly synthesis and offering exclusive biological benefits. This innovative nanomedicine technology shows great promise in medicine, providing a novel method for drug delivery and treatment. The advancement of biosynthesized nanomedicine has notable implications for the pharmaceutical and medical sectors, presenting a more precise and effective way to address diseases. This paper introduces a cost-effective, environmentally friendly, and dependable method for synthesizing silver and iron NPs. The method involves employing the aqueous leaf extract of Vallisneria spiralis Linnaeus. The biosynthesized NPs were analyzed utilizing several methods, including transmission electron microscope (TEM), dynamic light scattering (DLS), UV−visible spectroscopy, and ICP-MS.

Bacillus amyloliquefaciens TL promotes gut health of broilers by the contribution of bacterial extracellular polysaccharides through its anti-inflammatory potential.

The focal point of probiotic efficacy and a crucial factor influencing poultry cultivation lies in the level of intestinal inflammation. In conventional farming processes, the reduction of intestinal inflammation generally proves advantageous for poultry growth. This study investigated the impact of Bacillus amyloliquefaciens TL (B.A.-TL) on inflammatory factor expression at both tissue and cellular levels, alongside an exploration of main active secondary metabolites. The results demonstrated that broiler feeding with a basal diet containing 4 × 109 CFU/kg B.A.-TL markedly enhanced chicken growth performance, concomitant with a significant decrease in the expression of genes encoding inflammatory cytokines (e.g., CCL4, CCR5, XCL1, IL-1β, IL-6, IL-8, LITAF, and LYZ) in jejunum and ileum tissues. The extracellular polysaccharides of B.A.-TL (EPS-TL) exhibited notable suppression of elevated inflammatory cytokine expression induced by Escherichia coli O55 lipopolysaccharides (LPS) in chicken macrophage-like cells (HD11) and primary chicken embryonic small intestinal epithelial cells (PCIECs). Moreover, EPS-TL demonstrated inhibitory effect on NF-κB signaling pathway activation. These findings suggested that the metabolic product of B.A.-TL (i.e., EPS-TL) could partly mitigate the enhanced expression of inflammatory factors induced by LPS stimulation, indicating its potential as a key component contributing to the anti-inflammatory effects of B.A.-TL.

Development of GC–MS coupled to GC–FID method for the quantification of cannabis terpenes and terpenoids: Application to the analysis of five commercial varieties of medicinal cannabis

Cannabis terpenes and terpenoids are among the major classes of pharmacologically active secondary metabolites of therapeutic interest. Indeed, these hydrocarbon molecules, responsible for the characteristic aroma of cannabis flowers, are thought to be involved in a synergistic effect known as the “entourage effect”, together with cannabinoids. Numerous analytical studies have been carried out to characterize the terpene and terpenoid contents of some cannabis varieties, but they have not proposed any real quantification or have described a limited number of analytical standards or average response factors, which may have led to over- or underestimation of the real content of the cannabis flowers. Real and reliable quantification is necessary to justify the entourage effect. Here, we report a rigorous and precise GC–FID and GC–MS method for the identification and quantification of cannabis terpenes and terpenoids. This method is distinguished by the use of a high number of analytical standards, the determination of retention indices for all compounds studied, an exhaustive comparison of databases and scientific literature, the use of relevant response factors, and internal calibration for reliable results. It was applied to the study of terpenic compounds in five commercial varieties of medicinal cannabis produced by Bedrocan International: a CBD-rich (Bedrolite®), a THC/CBD balanced (Bediol®), and three THC-dominant (Bedrocan®, Bedica® and Bedrobinol®). Two extraction solvents are described (ethanol and hexane) to compare their selectivity towards target molecules, and to describe as exhaustively as possible the terpenic profile of the five pharmaceutical-grade varieties. Twenty-three standards were used for accurate dosages. This work highlights that the choice of solvent and the analysis method reliability are critical for the study of these terpenic compounds, regarding their contribution to the entourage effect.

Effect of Agitation and Temporary Immersion on Growth and Synthesis of Antibacterial Phenolic Compounds in Genus Drosera.

Sundews (Drosera sp.) are the source of biologically active secondary metabolites: phenolic acids, flavonoids, and 1,4-naphtoquinones. Because obtaining them from the natural environment is impossible (rare and endangered species), in this study modifications of traditional tissue cultures grown in solid medium (SM), such as agitated cultures (ACs) (cultures in liquid medium with rotary shaking) and temporary immersion bioreactors PlantformTM (TIB), were used for multiplication of four sundew species: Drosera peltata, Drosera indica, Drosera regia, and Drosera binata, with simultaneously effective synthesis of biologically active phenolic compounds. Each species cultivated on SM, AC, and TIB was tested for biomass accumulation, the content of total phenols and selected phenolic derivative concentrations (DAD-HPLC), the productivity on of phenolic compounds, as well as its antibacterial activity against two human pathogens: Staphylococcus aureus and Escherichia coli. The results showed that the type of culture should be selected for each species separately. Phytochemical analyses showed that the synthesis of secondary metabolites from the groups of phenolic acids, flavonoids, and 1,4-naphthoquinones can be increased by modifying the cultivation conditions. D. regia turned out to be the richest in phenolic compounds, including 1,4-naphtoquinones: plumbagin and ramentaceone. Extracts from D. indica and D. regia tissue showed strong antibacterial activity against both pathogens. It has also been shown that the growth conditions of sundews can modify the level of secondary metabolites, and thus, their biological activity.

Antagonism of rhizosphere Trichoderma brevicompactum DTN19 against the pathogenic fungi causing corm rot in saffron (Crocus sativus L.) in vitro.

Corm rot in saffron (Crocus sativus L.) significantly impacts yield and quality. Non-toxic fungi, particularly Trichoderma species, are valuable for biological control due to their production of diverse and biologically active secondary metabolites. This study aimed to isolate an effective antagonistic fungus against the pathogenic fungi causing corm rot in saffron. Four pathogenic fungi (Fusarium oxysporum, Fusarium solani, Penicillium citreosulfuratum, and Penicillium citrinum) were isolated from diseased saffron bulbs in Chongming. Initial screening through dual culture with these pathogens re-screening from rhizosphere soil samples of C. sativus based on its inhibitory effects through volatile, nonvolatile, and fermentation broth metabolites. The inhibitory effect of biocontrol fungi on pathogenic fungi in vitro was evaluated by morphological observation and molecular biology methods. Antagonistic fungi were identified as Trichoderma brevicompactum DTN19. F. oxysporum was identified as the most severe pathogen. SEM (scanning electron microscope) and TEM (transmission electron microscope) observations revealed that T. brevicompactum DTN19 significantly inhibited the growth and development of F. oxysporum mycelium, disrupting its physiological structure and spore formation. Additionally, T. brevicompactum DTN19 demonstrated nitrogen fixation and production of cellulase, IAA (Indole acetic acid), and siderophores. Whole-genome sequencing of strain DTN19 revealed genes encoding protease, cellulase, chitinase, β-glucosidase, siderophore, nitrogen cycle, and sulfate transporter-related proteins. T. brevicompactum DTN19 may inhibit the propagation of pathogenic fungi by destroying their cell walls or producing antibiotics. It can also produce IAA and iron carriers, which have the potential to promote plant growth. Overall, T. brevicompactum DTN19 showed the development prospect of biological agents.

Bryophyllum pinnatum (Lam.) Oken: unravelling therapeutic potential and navigating toxicity.

Bryophyllum pinnatum (Lam.) Oken, a multipurpose medicinal herb, has drawn much interest for its therapeutic qualities from both traditional and modern medicine systems. Many active secondary metabolites, such as bufadienolides, triterpenes, phenols, alkaloids, glycosides, lipids, flavonoids, and organic acids, are responsible for the plant's curative properties. B. pinnatum exhibits a noteworthy significance in oncological research by exhibiting its ability to modify numerous pathways, which may suggest a potential anticancer impact. The herb is recommended for treating lithiasis, a common cause of renal failure, due to its effectiveness in dissolving stones and avoiding crystal formation. The plant has a major impact on diabetes, especially type II diabetes. Moreover, the versatility of B. pinnatum extends to its examination in connection to COVID-19. However, caution is warranted, as B. pinnatum has been reported to possess toxicity attributed to the presence of bufadienolides in its metabolic profile. A comprehensive investigation is essential to thoroughly understand and confirm the synthesis of potentially hazardous compounds. This is crucial for minimizing their presence and ensuring the safe consumption of B. pinnatum among diverse populations of organisms. This review highlights the various medical uses of B. pinnatum, including its ability to effectively treat kidney and liver diseases, as well as its anti-leishmanial, neuropharmacological, antibacterial, immunosuppressive, anti-tumour, and cytotoxic effects. While extensively employed in both traditional and scientific domains, the plant's complete medicinal potential, molecular mechanisms, safety profile, and pharmacodynamics remain ambiguous, rendering it an ideal candidate for pioneering research endeavours.

Untargeted metabolic analysis of Epaltes mexicana by LC-QTOF-MS: Terpenes with activity against human cancer cell lines

Epaltes mexicana is a plant widely used in traditional medicine and as a food in Mexico; however, its phytochemical and pharmacological studies are limited. This study aimed to identify the active secondary metabolites of Epaltes mexicana and determine its cytotoxic activity on cancer cell lines. Three organic extracts were obtained by maceration using n-hexane, dichloromethane, and methanol. The n-hexane extract was fractioned by simple column chromatography. Eight terpenes were annotated in collection 6 (C6) by LC-QTOF-MS using a gradient elution and Electrospray Ionization (ESI) in positive ion mode: 1) Gibberellin A15, 2) farfugin A, 3) dehydromyodesmone, 4) eremopetasitenin A1, 5) hydroxyisonobilin, 6) anhydrocinnzeylanine, 7) nigakilactone H and 8) taxodione. On the other hand, C6 showed a concentration-dependent cytotoxic effect on cancer cell lines MCF-7 (Emax = 74.69 ± 6.19 % and IC50 = 6.31 μg/mL), MDA-MB-231 (Emax = 79.28 ± 12.12 % and IC50 = 124.21 μg/mL), and SiHa (Emax = 82.96 ± 6.02 % and IC50 = 124.31 μg/mL). The C6 did not show a cytotoxic effect against DU-145 and non-cancerous cells from the mammary glands MCF-10A. These results indicate cytotoxic specificity on cancer cell lines and support the hypothesis that terpenes identified in E. mexicana must be investigated and developed for non-clinical and clinical trials as potential anti-cancer drugs.

Solvent polarity effects on extraction yield, phenolic content, and antioxidant properties of Malvaceae family seeds: a comparative study

ABSTRACT The selection of appropriate solvents in the extraction process plays a crucial role in determining the nature and quantity of secondary metabolites extracted from medicinal plants. This study investigates the impact of solvent polarity on the extraction yield, phenolic content, and antioxidant properties of seeds from four plants belonging to the Malvaceae family: Abutilon fruticosum, Abutilon pannosum, Sida ovata and Malvastrum coromendelianum. The seed powder of each species was subjected to extraction using solvents with varying polarities, including acetone, chloroform, methanol, and water. Physicochemical analysis, extraction yield, quantitative phenolic content, and antioxidant activity were assessed. Total phenolic content (TPC) was determined using the Folin–Ciocalteu reagent, and antioxidant activity was measured using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. Analysis of variance (ANOVA) revealed that the choice of extracting solvents significantly influenced (p < 0.05) extraction yield, phenolic content, and antioxidant properties. In conclusion, water and methanol emerged as the most effective solvents for achieving high extraction yields, phenolic compound content, and free radical scavenging activities. The results demonstrated a robust correlation between total phenolic content (mg of Gallic acid equivalent/g of sample) and free radical scavenging activity (IC50). The study highlights the substantial impact of solvent type, suggesting the potential benefits of using a combination of polar and nonpolar solvents to enhance the extraction efficiency of phytochemicals, phenolic content and antioxidant quality from medicinal plant seeds. Future investigations should focus on isolating and identifying active secondary metabolites from these extracts.

In Vitro Evaluation of the Antimicrobial, Antioxidant, and Cytotoxicity Potential Coupled with Molecular Docking Simulation of the Dynamic Fermentation Characteristics of Marine-Derived Bacterium Halomonas saccharevitans

A variety of metabolites produced by marine microorganisms are gaining high value for their significant biological properties. Therefore, the search for active secondary metabolites from marine bacteria against pathogenic microorganisms has become essential for alternative and effective strategies. In this study, Halomonas saccharevitans, a marine-derived bacterium, was cultured and fermented on a liquid medium. The ethyl acetate (EtOAc) crude extract was then fractioned yielded five fractions to study their biological effect. Two fractions had significantly higher activity, i.e., absolute n-hexane against Staphylococcus aureus and Pseudomonas aeruginosa and absolute methanol (MeOH) against Escherichia coli and Bacillus subtilis, with promising MIC values. The time–kill kinetics assay for the very susceptible bacteria against active fractions was also examined. The antifungal assay of the active fractions had the highest activity against Aspergillus niger and Candida albicans with the examined variable MFC values. The cytotoxic assay against HepG2 cells showed promising activities, resulting in a 78% inhibition of cell viability. Moreover, the antioxidant activities showed reasonable inhibition values at 21.87 ± 0.85% and 98.25 ± 1.45%, compared to the control. Molecular docking revealed a high affinity between major detected compounds with free binding energies. The active fractions were characterized by the presence of diverse chemically esters, phenolics, essential oils, and other organic compounds detected by GC–MS. In conclusion, H. saccharevitans, derived from the Red Sea, might be useful as an alternative source for the possible production of bioactive substances with a variety of biomedical application.

Experimental Models in Unraveling the Biological Mechanisms of Mushroom-Derived Bioactives against Aging- and Lifestyle-Related Diseases: A Review.

Mushrooms have garnered considerable interest among researchers due to their immense nutritional and therapeutic properties. The presence of biologically active primary and secondary metabolites, which includes several micronutrients, including vitamins, essential minerals, and other dietary fibers, makes them an excellent functional food. Moreover, the dietary inclusion of mushrooms has been reported to reduce the incidence of aging- and lifestyle-related diseases, such as cancer, obesity, and stroke, as well as to provide overall health benefits by promoting immunomodulation, antioxidant activity, and enhancement of gut microbial flora. The multifunctional activities of several mushroom extracts have been evaluated by both in vitro and in vivo studies using cell lines along with invertebrate and vertebrate model systems to address human diseases and disorders at functional and molecular levels. Although each model has its own strengths as well as lacunas, various studies have generated a plethora of data regarding the regulating players that are modulated in order to provide various protective activities; hence, this review intends to compile and provide an overview of the plausible mechanism of action of mushroom-derived bioactives, which will be helpful in future medicinal explorations.

In-Silico Enactment of Musa paradisiaca (L.) Exudate Against the Urotlithiatic Disease by Docking Study

Objectives: Urolithiasis is a condition that occurs when the stones exit the renal pelvis and move onto the remainder of the urinary collecting system. Musa paradisiaca (L.) is a plant with high nutritive value and has been used for the treatment of various diseases including urolithiasis. The objective of the present study was designed to detect the receptor-ligand binding energy and interaction through a molecular docking from the bio-active compounds in an exudates of M. paradisiaca pseudo stem on urolithiatic causative protein named as glycolate oxidase receptor (PDB ID: 2RDU). Methods: In silico study especially molecular docking was performed using AutodockVina and the best confirmation between ligand and protein were selected using Lamarkaina Genetic Algorithm (LGA) and ligand protein interaction was visualized using PyMol viewer. Novelty: Totally three various bio-active compounds are elucidated named as Olean-12-ene-3 beta, 28-diol, Tricyclo[8.4.1.1(3,8)] hexadeca- 3,5,7,10,12,14-hexaene- 2,9-dione, anti and 2H-Pyran,2-(7-heptadecynyloxy)tetrahydro- from the exudate of experimental sample. Three phyto active compounds or secondary metabolites were used for the present prediction by docking. The present result of molecular docking clearly revealed that olean 12 -ene-3 beta, 28 diol is the best biologically effective ligand observed through a highest docking score (-7.2 k cal/mol) on glycolate oxidase receptor than other two ligand compounds. In conclusion the compound olean 12 -ene-3 beta, 28 diol could be act as a potential lead molecule for urolithiasis. Keywords: Musa paradisiaca, Exudate, GCMS, Urolithiasis, AutoDock, PyMol Viewer

Cytotoxic of Usnic Acid Isolated from Ramalina sp.

Ramalina sp. (Ramalinaceae) is a type of lichen known to contain many active secondary metabolite compounds that have the potential to be used as medicine or medicinal raw materials. One of the biological activities possessed by Ramalina sp. lichen is its anticancer activity. This research aims to isolate and identify active secondary metabolite compounds from the methanol extract of the Ramalina sp. lichen and to find out the cytotoxic activity of the isolated compound against MCF7 breast cancer cells. Compound 1 (usnic acid) was successfully isolated from fraction A. The isolation and purification process was carried out starting with a maceration process using acetone solvent, followed by silica gel column chromatography using a gradient solvent system consisting of n-hexane, n-hexane: EtOAc, EtOAc, EtOAc: MeOH, and MeOH with 5% increment of polarity to obtain 17 fractions (F-1 to F-17). From the 17 fractions obtained, fraction 3 (F-3) and fraction 4 (F-4) (eluted with n-hexane: EtOAc 30%), which had the same TLC profile, were combined and named as fraction A. Compound 1 (50 mg) is a yellow needle crystal that was formed in a bottle of fraction A, which was obtained after the process of combining fractions F-3 and F-4 and solvent evaporation process. The crystals were then separated and purified with CHCl3 and MeOH. Compound 1 was then characterized based on spectroscopic data. Various spectroscopic analysis data, including FTIR, 1D- and 2D-NMR, and LC-ESI-MS, show that Compound 1 is a dibenzofuran derivative compound with 18 carbons (3 from carbonyl groups (C=O) and 3 from methyl groups) and 2 hydroxyls (-OH). Cytotoxicity assay showed that at a low concentration of 18.75 ug/mL, Com-pound 1 caused a 67.06% decrease in MCF7 viability.

Microalgal Derivatives as Potential Nutraceutical and Pharmaceutical: Boon to Human Beings

Background: Marine resources have diverse biological and beneficial entities for human beings. Among them, microalgae are one of the eukaryotic photosynthetic organisms found in freshwater and marine environments with an immense source of metabolites. They consist of high nutraceutical and value-added compounds for health concerns. Objective: Most microalgal species like- chlorella, spirulina, Isochrysis, Nannochloropsis, etc. are found to synthesize biologically active secondary metabolites which are having high pharmaceutical and nutraceutical value. Some of the purely extracted compounds are Lecithin, fucoxanthin, astaxanthin, and most important Sulphur polysaccharides- fucose, galactose, xylose, rhamnose, etc. are providing anti-microbial, anti-fungal, anti-viral, anti-cancer and anti-diabetic activities. Methods: Many of the prior studies demonstrated the compilation of naturally derived secondary metabolites for their potential application in most fields. Because of their wide-ranging potential for use in biopharmaceutical and nutraceutical industries, microalgae have recently gained significant interest on a global scale. Result: Microalgae are both parts of the dietary ingredients and bioactive pharmaceuticals. A number of microalgal species have been explored for their significance towards their high-value products with their exceptional pharmacological and biological properties. Conclusion: This current review discussed the uses and benefits of microalgae for the manufacture of nutraceuticals and the medicinal use of diverse bioactive compounds.

Effects of montmorillonite on the loading and release of Piper betle L. extract applied for wound dressing based on chitosan/polyvinyl alcohol

In this study, we investigate the influence of montmorillonite (MMT) on the loading and release of Piper betle L. extract (PLE)–a medicinal herb containing active secondary metabolites with antibacterial, antioxidant, and anti–inflammatory effects. MMT (1 %, 3 %, 5 %) was blended into the chitosan/polyvinyl alcohol (CS/PVA) biocomposite film by the solution evaporation method, and then PLE was loaded onto this biocomposite using the immersion method. The tensile strength and the ability to absorb exudates of the CS/PVA film improved with the increase in MMT content. The MMT 3 % film was considered to have the best properties: good mechanical properties with a tensile strength of 27.44 ± 0.27 MPa and elongation at break of 14.57 ± 0.30 %, potential for wound dressing due to its ability to absorb wound exudate (swelling degree 61.70 ± 0.30 %) and a suitable water vapor transmission rate (1999 ± 47 g/m2·d). The presence of MMT (1 %, 3 %, 5 %) in the CS/PVA film led to an increase in the PLE loading efficiency of the films compared to the film without MMT, up to 1.65, 1.73, and 1.87 times, respectively. The MMT 3 % and 5 % films also exhibited a sustained PLE release effect for up to 24 h. MMT increased PLE bioavailability through bioactivity tests: antibacterial activity against both E. coli and S. aureus, antioxidant activity, effective healing of 2nd–degree burn wounds, and biocompatibility with the L929 fibroblasts cell line. The combination of physicochemical properties and biological activities proved that the MMT/PLE drug delivery system based on the CS/PVA biocomposite is promising for wound dressing.

Study the hepatoprotective effect of the methanolic extract of Plumbago auriculata Lam. against CCl4-induced hepatocyte damage in mice

The liver is an important organ in the body; its diseases are considered the major causes of morbidity and mortality around the world. Hepatotoxic chemicals cause damage to liver cells. Medicinal plants have a powerful hepatoprotective effect. This study has been aimed to assess the potential hepatoprotective effect of the methanolic extract of Plumbago auriculata Lam. against carbon tetrachloride (CCl4)- induced hepatocyte damage in albino mice. The methanolic extract of the plant was subjected to preliminary phytochemical analysis to determine the presence of secondary metabolites according to standard protocols. The acute toxicity study was carried out to measure the LD50. Swiss albino mice were divided into 4 groups treated intraperitoneally (IP) once daily for 7 days; the first group was a negative control, while the second group, considered as a CCl4 model, received a single dose of carbon tetrachloride during the last day. The third group received 500 mg/kg of body weight of methanolic extract of P. auriculata, and the last group received 100 mg/kg of body weight of standard silymarin, after 2 h of treatment, on the last day, all animals (except negative control group) have received CCl4 at a dose of 1 mL/kg of body weight. Biochemical analysis of collected blood and histopathological examination were performed. Results revealed that the preliminary phytochemical screening of the methanolic extract of the plant confirmed the presence of many biologically active secondary metabolites like flavonoids, steroids, terpenoids, saponins, glycosides and phenols and LD50 of the extract was 2.5 g/kg. The CCl4- intoxicated mice showed an increase in biochemical enzyme levels (ALT and AST) compared with the negative control group. The extract of 500 mg/kg revealed a significant reduction (p ? 0.05) in enzyme activities. The histopathological analysis of plant extract and silymarin-treated groups revealed a decrease in the pathological features compared with the CCl4-intoxicated group. It was concluded that the extract of P. auriculata has a significant effect on hepatoprotection against CCl4-induced hepatocyte damage and this may be because of the combined effects of the bioactive compositions of plant extract.

Marine seaweed endophytic fungi-derived active metabolites promote reactive oxygen species-induced cell cycle arrest and apoptosis in human breast cancer cells.

Endophytic fungi have an abundant sources rich source of rich bioactive molecules with pivotal pharmacological properties. Several studies have found that endophytic fungi-derived bioactive secondary metabolites have antiproliferative, anti-oxidant, and anti-inflammatory properties, but the molecular mechanism by which they induce cell cycle arrest and apoptosis pathways is unknown. This study aimed to determine the molecular mechanism underlying the anticancer property of the endophytic fungi derived active secondary metabolites on human breast cancer cells. In this study, we identified four endophytic fungi from marine seaweeds and partially screened its phytochemical properties by Chromatography-Mass Spectrometry (GC-MS) analysis. Moreover, the molecular mechanism underlying the anticancer property of these active secondary metabolites (FA, FB, FC and FE) on human breast cancer cells were examined on MCF-7 cells by TT assay, Apoptotic assay by Acridine orang/Ethidium Bromide (Dual Staining), DNA Fragmentation by DAPI Staining, reactive oxygen species (ROS) determination by DCFH-DA assay, Cell cycle analysis was conducted Flow cytometry and the apoptotic signalling pathway was evaluated by westernblot analysis. Doxorubicin was used as a positive control drug for this experiment. The GC-MS analysis of ethyl acetate extract of endophytic fungi from the marine macro-algae revealed the different functional groups and bioactive secondary metabolites. From the library, we observed the FC (76%), FB (75%), FA (73%) and FE (71%) have high level of antioxidant activity which was assessed by DPPH scavenging assay. Further, we evaluated the cytotoxic potentials of these secondary metabolites on human breast cancer MCF-7 cells for 24h and the IC50 value were calculated (FA:28.62 ± 0.3µg/ml, FB:49.81 ± 2.5µg/ml, FC:139.42 ± µg/ml and FE:22.47 ± 0.5µg/ul) along with positive control Doxorubicin 15.64 ± 0.8µg/ml respectively by MTT assay. The molecular mechanism by which the four active compound induced apoptosis via reactive oxygen species (ROS) and cell cycle arrest in MCF-7 cells was determined H2DCFDA staining, DAPI staining, Acridine orange and ethidium bromide (AO/EtBr) dual staining, flowcytometry analysis with PI staining and apoptotic key regulatory proteins expression levels measured by westernblot analysis. Our findings, revealed the anticancer potential of endophytic fungi from marine seaweed as a valuable source of bioactive compounds with anticancer properties and underscore the significance of exploring marine-derived endophytic fungi as a promising avenue for the development of novel anticancer agents. Further investigations are necessary to isolate and characterize specific bioactive compounds responsible for these effects and to validate their therapeutic potential in preclinical and clinical settings.