In silico functional and structural characterization revealed virulent proteins of Francisella tularensis strain SCHU4.

Francisella tularensis is a pathogenic, aerobic gram-negative coccobacillus bacterium. It is the causative agent of tularemia, a rare infectious disease that can attack skin, lungs, eyes, and lymph nodes. The genome of F. tularensis has been sequenced, and ~16% of the proteome is still uncharacterized. Characterizations of these proteins are essential to find new drug targets for better therapeutics. In silico characterization of proteins has become an extremely important approach to determine the functionality of proteins as experimental functional elucidation is unable to keep pace with the current growth of the sequence database. Initially, we have annotated 577 Hypothetical Proteins (HPs) of F. tularensis strain SCHU4 with seven bioinformatics tools which characterized them based on the family, domain and motif. Out of 577 HPs, 119 HPs were annotated by five or more tools and are further screened to predict their virulence properties, subcellular localization, transmembrane helices as well as physicochemical parameters. VirulentPred predicted 66 HPs out of 119 as virulent. These virulent proteins were annotated to find the interacting partner using STRING, and proteins with high confidence interaction scores were used to predict their 3D structures using Phyre2. The three virulent proteins Q5NH99 (phosphoserine phosphatase), Q5NG42 (Cystathionine beta-synthase) and Q5NG83 (Rrf2-type helix turn helix domain) were predicted to involve in modulation of cytoskeletal and innate immunity of host, H2S (hydrogen sulfide) based antibiotic tolerance and nitrite and iron metabolism of bacteria. The above predicted virulent proteins can serve as novel drug targets in the era of antibiotic resistance.

Cardioprotective Activity of Cassia fistula L. Bark Extract in Isoproterenol-Induced Myocardial Infarction Rat Model.

Cassia fistula Linn, generally recognized as Indian laburnum, is one of the ancient trees in the Indian subcontinent used for its ornamental and diverse medicinal properties. It is known for its ethnic medicinal uses in inflammatory and infectious pathologies such as antihelmintic, purgative, carminative, antipyretic, expectorant, analgesic, laxative, antiseptic, and antidote against snake poison. The Cassia bark is rich in anthraquinones, flavanols glycosides, and sitosterols, which renders it cardioprotective properties. The existing experiments were designed to assess the potential of Cassia fistula bark against isoproterenol (ISP)-induced cardiotoxicity in rats, which has not been validated yet. The bark was successively extracted with five different solvents, and each extract was subjected to in vitro antioxidant studies. Further acute oral toxicity assays were carried out preceding in vivo myocardial studies. Cardiotoxicity-inducing agent, ISP, was administrated to the rats for two consecutive days (8th and 9th). Based on in vitro studies, the Cassia fistula methanolic extract (CFME) was administered in two doses: CFME-LD (lower dose 250 mg/kg) and CFME-HD (high dose 500 mg/kg) separately. It was found that CFME produced a substantial decrease in lipid peroxidation and an increase in antioxidants in myocardial tissues. CFME abrogated the levels of triglyceride and total cholesterol with a decrease in alanine transaminase (ALT) and aspartate transaminase (AST) activity in serum at both doses. 2,3,5-Triphenyltetrazolium chloride (TTC) staining and histopathology also revealed the protective effects of CFME against ISP-induced myocardial infarction. The study showed the significant role of the CFME as a strong antioxidant and cardioprotective action in ISP-induced toxicity.

Biomarker Analysis Based Chemoprofiling of Polyherbal Ayurvedic Formulation Containing Vitis vinifera L. by Validated UPLC-MS/MS Method.

BACKGROUND: Drakshasava is one of the commercial Ayurvedic medicines from India, prepared from grapes and spices. It is believed to address health imbalances and claimed to be beneficial for weakness, bleeding disorders, and various inflammatory diseases. It has been reported to possess pharmacological activities such as diuretic, cardioprotective, and antimicrobial. Being a polyherbal mixture, it faces challenges in its standardization and quality control. OBJECTIVE: The aim of the present study is to develop a validated UPLC-MS/MS method for simultaneous quantification of 10 polyphenolic biomarkers in Drakshasava. It explores the effect of Vitis vinifera L. and additional herbs on fermentation with respect to bioactive compounds through the successive addition method. METHODS: The MS methods were optimized in multiple-reaction monitoring (MRM) mode with ESI while chromatographic separation was achieved on an Acquity UPLC BEH C18 column using both isocratic and gradient elution in water and acetonitrile containing 0.1% formic acid. RESULTS: The developed method was validated as per ICH-Q2B guidelines and found to be within the assay variability limits. Gallic acid was found to be the most abundant marker in all the samples followed by resveratrol. The content of all the markers has been found to be increased significantly post-fermentation, compared to decoction except kaempferol. The successive addition of prashpeka drvya (minor herbs) in the formulation showed variability at different stages with respect to the selected markers and did not exhibit major changes in the chemical profiling of the final product. CONCLUSION: The developed method was found to be rapid, accurate, reliable, and highly sensitive for the simultaneous quantification of selected biomarkers in Drakshasava. The research is the first chemometric report on the standardization of Drakshasava by validated UPLC-MS/MS method. It may prove to be a useful tool for the development of new phytopharmaceutical drugs and further quality control of other polyherbal formulations.