Curvuluria lunata is a melanized fungus pathogenic to both plants and animals including humans, causing from mild, febrile to life-threatening illness if not well treated. In humans, it is an etiological agent of keratomycosis, sinusitis, and onychomycosis in immunocompromised and immunocompetent patients. The development of multiple-drug-resistant strains poses a critical treatment issue as well as public health problem. Natural products are attractive prototypes for drug discovery due to their broad-spectrum efficacy and lower side effects. The present study explores possible targets of natural antifungal compounds (α-pinene, eugenol, berberine, and curcumin) against C. lunata via gene expression analysis, molecular docking interaction, and molecular dynamics (MD) studies. Curcumin, berberine, eugenol, and α-pinene exhibited in vitro antifungal activity at 78 μg/ml, 156 μg/ml, 156 μg/ml, and 1250 μg/ml, respectively. In addition, treatment by these compounds led to the complete inhibition of conidial germination and hindered the adherence when observed on onion epidermis. Several pathogenic factors of fungi are crucial for their survival inside the host including those involved in melanin biosynthesis, hyphal growth, sporulation, and mitogen-activated protein kinase (MAPK) signalling. Relative gene expression of velB, brn1, clm1, and pks18 responsible for conidiation, melanin, and cell wall integrity was down-regulated significantly. Results of molecular docking possessed good binding affinity of compounds and have confirmed their potential targets as THR and VelB proteins. The docked structures, having good binding affinity among all, were further refined, and rescored from their docked poses through 100-ns long MD simulations. The MDS study revealed that curcumin formed a stable and energetically stabilized complex with the target protein. Therefore, the study concludes that the antifungal compounds possess significant efficacy to inhibit C. lunata growth targeting virulence proteins/genes involved in spore formation and melanin biosynthesis.

In this study, miRNA profiling of cells infected with lumpy skin disease virus (LSDV) was conducted for the first time. As compared to the mock-infected cells, LSDV-infected primary lamb testicle (LT) cells revealed dysregulation of 64, 85, and 85 miRNAs at 12 hours post-infection (hpi), 48 hpi and 72 hpi, respectively. While some of these miRNAs were found to be specifically dysregulated at a particular time point following LSDV infection, others were commonly dysregulated across all three time points. The analysis of the differentially expressed miRNA-mRNA interaction networks, Gene ontology analysis of the predicted targets and KEGG analysis of the highly enriched pathways revealed several cellular factors/pathways involved in protein/ion/enzyme binding, cell differentiation, movement of subcellular component, calcium reabsorption, aldosterone synthesis and secretion and, melanogenesis. Besides understanding virus replication, virus-host interactions and disease pathogenesis, these miRNAs and their cellular targets may serve as biomarkers as well as novel targets for therapeutic intervention against LSDV.