Knowledge on interactions between earthworms and microbes at molecular level has been obscure. Mechanism by which earthworms defend themselves against different microorganisms in the soil is largely unknown. Coelomic fluid from earthworm species has been shown to have antimicrobial activity. In this study we report that coelomic fluid isolated from Eudrilus eugeniae has antifungal activity against four different phytopathogens such as Rhizoctonia solani, Fusarium oxysporum, Aspergillus flavus and Verticillium dahlia. Incubation of coelomic fluid with fungal hyphae of all these four strains for 12 hours revealed that the growth of hyphae was greatly inhibited. Light microscopy studies confirmed that the hyphae growth was greatly reduced by incubation of coelomic fluid. Analysis of the fluid in the microscope showed the presence of different types of coelomocytes in the mucous fluid. Other molecules and compounds in the coelomic fluid might also have played role in antifungal activity.
To identify antimitotic compounds from abundant and inexpensive plant resources, banana pseudo-stem was (BPS) chosen. Onion root tip assay and earthworm regeneration assay were carried out to test theantimitotic potential of aqueous extract of BPS.Earthworm (Eudrilus eugeniae) regeneration assay exploits the regeneration ability of amputated earthworms that retain the clitellum region. Aqueous extract of BPS decreased the mitotic index in Allium cepa root tips. Besides, thisaqueous extract of BPS inhibited the regeneration of blastema from amputated earthworms as well. Validation of this extract with MTT (3-(4,5-dimethyl thiazolyl-2-yl)- 2,5-diphenyltetrazolium bromide) assay using MCF-7 breast cancer cell linefurther supported the presence of antimitotic compounds. Aqueous BPS extract was further fractionated with ethyl acetateand it was found to inhibit the regeneration of new tissues from amputated earthworms. Liquid Chromatography and Mass spectrometry (LC-MS) analysis was performed with aqueousBPS extract to predict the lead compounds.Prediction analysis with mass values revealed the presence of three different compounds viz. α-tocotrienol, 1,2,4-nonadecanetriol and 3',4',7-trihydroxyisoflavone, which were already reported to inhibit the cell division. All our results strongly supported that banana pseudo-stem extract possesses antimitotic compounds. This is the first report of identification of putative antimitotic compounds from aqueous extract of Musa paradisiaca var. Robusta by using earthworms.
Heat shock protein 60 kDa (HSP60) is a crucial chaperone that guides appropriate folding of denatured proteins under heat stress conditions. HSP60 provides assistance in correct folding of a multitude of denatured proteins. The Group II eukaryotic chaperonin TCP-1 ring complex is (TRiC or CCT) important cytosolic chaperonin which plays important role in folding of essential subset of cytosolic proteins and is believed to be highly conserved among different eukaryotic species. In this study, an extensive in silico analysis has been performed ranging from sequence comparison among species to homology modeling of Bos taurus Cct5 protein and determination of Hsp60 interacting partners. The comparative analysis of the protein revealed certain significant variations in Bos taurus. The experimental protein structure for Cct5 in Bos taurus is unknown till date, therefore the putative protein structure was determined using homology modeling. The stereo chemical properties of protein were assessed by utilizing several scrutiny tools such as PROCHECK, VERIFY3D and RAMPAGE to ensure model accuracy. The mode of interaction between HSF1 and Cct5 protein was evaluated by molecular docking studies.
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