Phytochemical Screening and Bioactivity Studies of Endophytes Cladosporium sp. Isolated from the Endangered Plant Vateria Indica Using In Silico and In Vitro Analysis.

Vateria indica is persistent tree used in Unani sources for the medication and classified as critically endangered. Thus, endophytes for alternative methods to explore these endangered Plants having rich source pharmaceuticals' active molecules for drug development and production. Endophytes comprises unexplored microbes as a potential source of rich pharmaceutically bioactive compounds attributable to their relationship with the host. In the current study, we have isolated endophyte fungi Cladosporium from the plant Vateria indica and performed phytochemical screening of its ethanolic extract to detect the phytochemicals using thin layer chromatography (TLC), gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography (HPLC), UV-visible spectrophotometry (UV-VIS), and Fourier transform infrared spectroscopy (FTIR). GC-MS analysis revealed the presence of an anticancer compound hydroxymethyl colchicine, antioxidant compound benzoic acid, and antimicrobial 2-(4-chlorophenoxy)-5-nitro in endophyte fungal extract of plant Vateria indica. Moreover, in silico analysis of bioactive compounds identified by GC-MS analysis using the Autodock Vina and SwissADME confirmed excellent anticancer activity methanone, [4-amino-2-[(phenylmethyl) amino]-5-thiazolyl] (4-fluorophenyl)- and hydroxymethyl colchicine against 6VO4 (Bfl-1 protein) as per Lipinski rule. Furthermore, we also demonstrated the excellent antioxidant of endophytic extract compared to plant extract by DPPH and ABTS assay, as well as antimicrobial activity against both Gram (+ ve) and Gram (- ve) bacteria. Moreover, the endophytic extract also showed its antimitotic activity with a mitotic index of 65.32, greater than the plant extract of 32.56 at 10 mg/ml. Thus endophytic fungi Cladosporium species isolated from plant Vateria indica might be used as a potential source for phytochemical anticancer hydroxymethyl colchicine, an antioxidant benzoic acid, and antimicrobial 2-(4-chlorophenoxy)-5-nitro.

Isolation and Phytochemical Screening of Endophytic Fungi Isolated from Medicinal Plant Mappia foetida and Evaluation of Its In Vitro Cytotoxicity in Cancer.

Isolated endophyte fungi from Mappia foetida have been explored as a potential source for the mass production of anticancer drug lead compounds in the current study. Since medical plants are not feasible economically for mass production of bioactive pharmaceutical important molecules using plant tissue culture due to factors like media design and fungal contamination, endophyte fungal mass culture have been an alternative for the relatively easy and inexpensive production. Two endophytic fungi isolated, Alternaria alternata and Fusarium species were mass cultured and their prepared alcoholic extract subjected to standard procedures to identify the phytochemical screening by gas chromatography-mass spectrometry (GCMS), high-performance liquid chromatography (HPLC), UV visible spectrophotometry (UV-VIS), and Fourier transform infrared spectroscopy (FTIR). GC-MS analysis revealed the presence of three major compounds in the extracts. The phytochemical screening confirmed the presence of an anticancer compound (camptothecin) in their extract. Moreover, the dose-dependent anticancer activity of ethanol extract was demonstrated against cervical carcinoma (HeLa), breast carcinoma (MCF-7), non-small cell lung carcinoma (H1975), and hepatocellular carcinoma cell line (Hep G2) by MTT assay where doxorubicin was used as the positive control. Furthermore, the microscopic examination also confirmed the cytotoxic effect of extract of endophytic fungi Alternaria alternata and Fusarium species against tested cancer cells. Hence, endophytic fungi Alternaria alternata and Fusarium species might be exploited for mass production of phytochemicals having anticancer activity.

Apoptotic Effect and Anticancer Activity of Biosynthesized Silver Nanoparticles from Marine Algae Chaetomorpha linum Extract Against Human Colon Cancer Cell HCT-116.

The green approach of nanoparticle synthesis has gained more attention by researchers because of its nontoxic, eco-friendly, biocompatible, and sustainable nature. The present research investigated the anticancer effectiveness of silver nanoparticles synthesized from marine algae Chaetomorpha linum (C. linum) against colon cancer cell HCT-116 in vitro. Biosynthesized silver nanoparticles (C-AgNPs) are characterized using UV-spectrophotometry, dynamic light scattering (DLS), X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). We demonstrated the dose-dependent cytotoxic effect of C-AgNPs in human colorectal carcinoma cells (HCT-116) using MTT assay. The apoptosis induction in HCT-116 cells caused by C-AgNPs has studied fluorescence microscope by staining with fluorogenic agents 4',6-diamidino-2-phenylindole (DAPI), rhodamine 123, and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA). By using a flow cytometric test, the apoptotic action of C-AgNPs was performed. The immunoblotting study of caspases, as well as pro-apoptotic and anti-apoptotic protein expression, was studied using the PCR technique to understand the underlying molecular mechanism of C-AgNPs on cancer cells. The apoptotic studies showed an increase in the expression of apoptotic caspase 3, caspase 9, BH3-interacting domain death agonist (Bid), and Bax, along with a decrease in the anti-apoptotic protein like Bcl-2 and Bcl-xl, thereby veritably confirmed by immunoblotting and qPCR technique. The biosynthesized C-AgNPs was an efficient anticancer agent that can induce apoptosis in the HCT-116 colon cells.