Identification of Small-Molecule Inhibitors of the Salmonella FraB Deglycase Using a Live-Cell Assay.

Nontyphoidal salmonellosis is one of the most significant foodborne diseases in the United States and globally. There are no vaccines available for human use to prevent this disease, and only broad-spectrum antibiotics are available to treat complicated cases of the disease. However, antibiotic resistance is on the rise and new therapeutics are needed.

Demonstrating the utility of sugar-phosphate phosphatases in coupled enzyme assays: galactose-1-phosphate uridylyltransferase as proof-of-concept.

During our biochemical characterization of select bacterial phosphatases belonging to the haloacid dehalogenase superfamily of hydrolases, we discovered a strong bias of Salmonella YidA for glucose-1-phosphate (Glc-1-P) over galactose-1-phosphate (Gal-1-P). We sought to exploit this ability of YidA to discriminate these two sugar-phosphate epimers in a simple coupled assay that could be a substitute for current cumbersome alternatives. To this end, we focused on Gal-1-P uridylyltransferase (GalT) that is defective in individuals with classical galactosemia, an inborn disorder.

Serendipitous Discovery of a Competitive Inhibitor of FraB, a Deglycase and Drug Target.

Although salmonellosis, an infectious disease, is a significant global healthcare burden, there are no -specific vaccines or therapeutics for humans. Motivated by our finding that FraB, a deglycase responsible for fructose-asparagine catabolism, is a viable drug target, we initiated experimental and computational efforts to identify inhibitors of FraB. To this end, our recent high-throughput screening initiative yielded almost exclusively uncompetitive inhibitors of FraB.

Use of tandem affinity-buffer exchange chromatography online with native mass spectrometry for optimizing overexpression and purification of recombinant proteins.

Purification of recombinant proteins typically entails overexpression in heterologous systems and subsequent chromatography-based isolation. While denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis is routinely used to screen a variety of overexpression conditions (e.g.

Sugar-Phosphate Toxicities.

Accumulation of phosphorylated intermediates during cellular metabolism can have wide-ranging toxic effects on many organisms, including humans and the pathogens that infect them. These toxicities can be induced by feeding an upstream metabolite (a sugar, for instance) while simultaneously blocking the appropriate metabolic pathway with either a mutation or an enzyme inhibitor. Here, we survey the toxicities that can arise in the metabolism of glucose, galactose, fructose, fructose-asparagine, glycerol, trehalose, maltose, mannose, mannitol, arabinose, and rhamnose.

Free radical scavenging activity and comparative metabolic profiling of in vitro cultured and field grown Withania somnifera roots.

Free radical scavenging activity (FRSA), total phenolic content (TPC), and total flavonoid content (TFC) of in vitro cultured and field grown Withania somnifera (Ashwagandha) roots were investigated. Withanolides analysis and comprehensive metabolic profiling between 100% methanol extracts of in vitro and field grown root tissues was performed using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectrometry (GC-MS), respectively. Significantly higher levels of FRSA, TPC, and TFC were observed in in-vitro cultured roots compared with field grown samples.

Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis.

Background: The production of metabolites via in vitro culture is promoted by the availability of fully defined metabolic pathways. Withanolides, the major bioactive phytochemicals of Withania somnifera, have been well studied for their pharmacological activities. However, only a few attempts have been made to identify key candidate genes involved in withanolide biosynthesis.

Antipyretic, antinociceptive and anti-inflammatory activity of Premna herbacea roots.

The alcoholic extract of the roots of Premna herbacea was investigated for its antipyretic, antinociceptive and anti-inflammatory potential in animal models. The extract, when administered orally to mice has been found to be safe up to a dose of 8.0 g/kg.

Antinociceptive, anti-inflammatory and antipyretic effects of ethanol extract of Clerodendron serratum roots in experimental animals.

The alcoholic extract (50, 100 and 200 mg/kg, p.o) of Clerodendron serratum roots produced a significant antinociceptive, anti-inflammatory and antipyretic activities in animal models. The results support the traditional claims of C.

A study on the role of cholinergic and gamma amino butyric acid systems in the anti-nociceptive effect of gossypin.

1. The participation of cholinergic and gamma amino butyric acid (GABA) neurotransmitter systems in the anti-nociceptive effect of gossypin was investigated using pharmacological tools. 2.

Analgesic activity of certain flavone derivatives: a structure-activity study.

1. Flavone, its methoxy derivatives and flavanone were synthesized by standard methods and were tested for analgesic activity in mice by employing acetic acid writhing and tail flick methods. 2.

Anti-inflammatory and mast cell protective effect of ficus religiosa.

The aqueous extract of bark of Ficus religiosa was prepared and investigated for its anti-inflammatory effect and for its protective effect on mast cells against degranulation. A significant anti-inflammatory effect was observed in both acute and chronic models of inflammation. The extract also protected mast cells from degranulation induced by various degranulatiors.

Analgesic activity of certain flavone derivatives: a structure-activity study.

Flavone and 10 hydroxy and glucoside flavone derivatives were synthesised. They were tested for their analgesic effect in mice employing acetic acid-induced writhing and tail immersion methods. Subcutaneously all the tested compounds exhibited significant analgesic activity with varying potencies in both assay models.

Identification of the ingredients in curna, kvatha curna lehya and rasayana - a simple microscopic method.

Triphala Curna, Triphatladi Kvatha Curna, Inji Rasayanam and Manibhadra Lehya of Indian System of Medicine were examined microscopically and the methods of identifying their ingredients were reported as one of the quality control standards.