Exploring the antibacterial, antibiofilm, and antivirulence activities of tea tree oil-containing nanoemulsion against carbapenem-resistant Serratia marcescens associated infections.

Carbapenem-resistant Serratia marcescens (CRE-S. marcescens) has recently emerged as an opportunistic human pathogen that causes various nosocomial and respiratory tract infections. The prognosis for CRE-S. marcescens-related infections is very poor and these infections are difficult to treat. This study investigated the synthesis of tea tree oil nanoemulsion (TTO-NE) and its impact on CRE-S. marcescens both in vitro and in vivo. TTO-NE was characterized by dynamic light scattering (DLS) and effectively eradicated bacterial planktonic and sessile forms, reduced bacterial virulence factors, and generated reactive oxygen species (ROS) in the bacterial cell. Notably, TTO-NE was efficient in reducing the colonization of CRE-S. marcescens in a C. elegans in vivo model. The data suggest that TTO-NE might be an excellent tool to combat infections associated with CRE-S. marcescens.

Antibacterial, anti-biofilm, and anti-virulence potential of tea tree oil against leaf blight pathogen Xanthomonas oryzae pv. oryzae instigates disease suppression.

Bacterial leaf blight (BLB) disease, caused by Xanthomonas oryzae pv. oryzae (Xoo), causes major annual economic losses around the world. Inorganic copper compounds and antibiotics are conventionally used to control BLB disease. They often cause environmental pollution, contributing to adverse effects on human health. Therefore, research is now leading to the search for alternative control methods. Tea tree oil (TTO) is obtained from a traditional medicinal plant, Melaleuca alternifolia, with antibacterial properties. In this study, we found that TTO showed antibacterial activity against Xoo with a minimum inhibitory concentration (MIC) of 18 mg/ml. These antagonistic activities were not limited only to planktonic cells, as further studies have shown that TTO effectively eradicated sessile cells of Xoo in both initial and mature biofilms. Furthermore, it was also observed that TTO reduced various key virulence properties of Xoo, such as swimming, swarming motility, and the production of extracellular polymeric substances, xanthomonadin, and exoenzymes. TTO triggered ROS generation with cell membrane damage as an antibacterial mode of action against Xoo. The in silico study revealed that 1,8-cineole of TTO was effectively bound to two essential proteins, phosphoglucomutase and peptide deformylase, responsible for the synthesis of EPS and bacterial survival, respectively. These antibacterial and anti-virulence activities of TTO against Xoo were further confirmed by an ex vivo virulence assay where TTO significantly reduced the lesion length caused by Xoo on rice leaves. All these data concluded that TTO could be a safe, environment-friendly alternative approach for the comprehensive management of BLB disease.

Sodium alginate-based edible coating containing nanoemulsion of Citrus sinensis essential oil eradicates planktonic and sessile cells of food-borne pathogens and increased quality attributes of tomatoes.

There is a growing interest from the worldwide scientific community in formulating edible- biodegradable coatings to replace non-biodegradable and expensive commercial wax-based coatings for preserving postharvest quality attributes of vegetables including tomatoes. Postharvest tomatoes are a suspected vehicle for both Salmonella and Listeria in food poisoning incidents. In this work, the effectiveness of edible nano-emulsion coatings containing sweet orange essential oil and sodium alginate were prepared and characterized, then evaluated antibacterial and antibiofilm activity against Salmonella and Listeria and simultaneously, examined its coating effect on various quality characteristics of tomatoes at 22 ± 2 °C over a 15 days storage period. DLS (Dynamic light scattering) study revealed stable nanoemulsion formulation with 43.23 nm particle size. The high whiteness index of nanoemulsion has a positive impact on product marketability and desirability. Antibacterial and antibiofilm studies revealed nanoemulsion effectively eradicate both sessile and planktonic forms of Salmonella and Listeria in both single and multi-species culture conditions. Tomatoes coated with edible coating significantly enhanced firmness up to 33%, decreased total mesophilic bacteria including Salmonella and Listeria, and reduced weight loss up to 3 fold lower than uncoated one. Sensory analysis revealed that the use of the edible coating increased the total acceptance scores of tomatoes.

Azadiradione ameliorates polyglutamine expansion disease in Drosophila by potentiating DNA binding activity of heat shock factor 1.

Aggregation of proteins with the expansion of polyglutamine tracts in the brain underlies progressive genetic neurodegenerative diseases (NDs) like Huntington's disease and spinocerebellar ataxias (SCA). An insensitive cellular proteotoxic stress response to non-native protein oligomers is common in such conditions. Indeed, upregulation of heat shock factor 1 (HSF1) function and its target protein chaperone expression has shown promising results in animal models of NDs. Using an HSF1 sensitive cell based reporter screening, we have isolated azadiradione (AZD) from the methanolic extract of seeds of Azadirachta indica, a plant known for its multifarious medicinal properties. We show that AZD ameliorates toxicity due to protein aggregation in cell and fly models of polyglutamine expansion diseases to a great extent. All these effects are correlated with activation of HSF1 function and expression of its target protein chaperone genes. Notably, HSF1 activation by AZD is independent of cellular HSP90 or proteasome function. Furthermore, we show that AZD directly interacts with purified human HSF1 with high specificity, and facilitates binding of HSF1 to its recognition sequence with higher affinity. These unique findings qualify AZD as an ideal lead molecule for consideration for drug development against NDs that affect millions worldwide.

Smokeless tobacco extract (STE)-induced toxicity in mammalian cells is mediated by the disruption of cellular microtubule network: a key mechanism of cytotoxicity.

Smokeless tobacco usage is a growing public health problem worldwide. The molecular mechanism(s) underlying smokeless tobacco associated tissue damage remain largely unidentified. In the present study we have tried to explore the effects of aqueous extract of smokeless tobacco (STE) on tubulin-microtubule, the major cytoskeleton protein that maintains cells morphology and participates in cell division. Exposure to STE resulted in dose-dependent cytotoxicity in a variety of mammalian transformed cell lines such as human lung epithelial cells A549, human liver epithelial cells HepG2, and mouse squamous epithelial cells SCC7, [corrected] as well as non-tumorogenic human peripheral blood mononuclear cells PBMC. Cellular morphology of STE-treated cells was altered and the associated disruption of microtubule network indicates that STE targets tubulin-microtubule system in both cell lines. Furthermore it was also observed that STE-treatment resulted in the selective degradation of cellular tubulin, whereas actin remains unaltered. In vitro, polymerization of purified tubulin was inhibited by STE with the IC50 value∼150 µg/ml and this is associated with the loss of reactive cysteine residues of tubulin. Application of thiol-based antioxidant N-acetyl cysteine (NAC) significantly abrogates STE-mediated microtubule damage and associated cytotoxicity in both A549 and HepG2 cells. These results suggest that microtubule damage is one of the key mechanisms of STE-induced cytotoxity in mammalian cells.