An In Silico Molecular Modelling-Based Prediction of Potential Keap1 Inhibitors from Hemidesmus indicus (L.) R.Br. against Oxidative-Stress-Induced Diseases.

The present study investigated the antioxidant potential of aqueous methanolic extracts of Hemidesmus indicus (L.) R.Br., followed by a pharmacoinformatics-based screening of novel Keap1 protein inhibitors. Initially, the antioxidant potential of this plant extract was assessed via antioxidant assays (DPPH, ABTS radical scavenging, and FRAP). Furthermore, 69 phytocompounds in total were derived from this plant using the IMPPAT database, and their three-dimensional structures were obtained from the PubChem database. The chosen 69 phytocompounds were docked against the Kelch-Neh2 complex protein (PDB entry ID: 2flu, resolution 1.50 Å) along with the standard drug (CPUY192018). H. indicus (L.) R.Br. extract (100 µg × mL-1) showed 85 ± 2.917%, 78.783 ± 0.24% of DPPH, ABTS radicals scavenging activity, and 161 ± 4 µg × mol (Fe (II)) g-1 ferric ion reducing power. The three top-scored hits, namely Hemidescine (-11.30 Kcal × mol-1), Beta-Amyrin (-10.00 Kcal × mol-1), and Quercetin (-9.80 Kcal × mol-1), were selected based on their binding affinities. MD simulation studies showed that all the protein-ligand complexes (Keap1-HEM, Keap1-BET, and Keap1-QUE) were highly stable during the entire simulation period, compared with the standard CPUY192018-Keap1 complex. Based on these findings, the three top-scored phytocompounds may be used as significant and safe Keap1 inhibitors, and could potentially be used for the treatment of oxidative-stress-induced health complications.

Biocatalytic and antibacterial visualization of green synthesized silver nanoparticles using Hemidesmus indicus.

In the present investigation, we described the green synthesis of silver nanoparticles using plant leaf extract of Hemidesmus indicus. The synthesized silver nanoparticles were characterized by UV-visible spectroscopy, fourier transform infra-red spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDX). TEM images proved that the synthesized silver nanoparticles were spherical in shape with an average particle size of 25.24 nm. To evaluate antibacterial efficacy, bacteria was isolated from poultry gut and subjected to 16S rRNA characterization and confirmed as Shigella sonnei. The in vitro antibacterial efficacy of synthesized silver nanoparticles was studied by agar bioassay, well diffusion and confocal laser scanning microscopy (CLSM) assay. The H. indicus mediated synthesis of silver nanoparticles shows rapid synthesis and higher inhibitory activity (34 ± 0.2 mm) against isolated bacteria S. sonnei at 40 µg/ml.

Shikimate pathway modulates the elicitor-stimulated accumulation of fragrant 2-hydroxy-4-methoxybenzaldehyde in Hemidesmus indicus roots.

Enzymatic route to fragrant 2-hydroxy-4-methoxybenzaldehyde (MBALD) formation in Hemidesmus indicus roots is not known. Earlier studies with H. indicus excised roots suggested a possible origin of MBALD via central phenylpropanoid pathway. Different elicitors (e.g., chitosan, methyl jasmonate, yeast extract) were tested for their relative efficiency in uplifting MBALD accumulation in roots, amongst which, treatment with yeast extract for 18 h showed maximum accumulation in excised roots. As benzoate pathways originate either directly from shikimate or via phenylpropanoid pathway, this study aimed at finding the roles of shikimate pathway in uplifting/enhancing MBALD accumulation in H. indicus roots upon elicitation. In fact, a sharp increase in shikimate dehydrogenase (SKDH; E.C. 1.1.1.25) along with phenylalanine ammonia-lyase (PAL; E.C. 4.3.1.24) activities was noted on a time-course basis in yeast extract-treated roots as compared to the untreated ones. PAL as well as phenylpropanoid C2 side-chain cleavage activities (leading to p-hydroxybenzaldehyde, the first benzoate product formed in the MBALD pathway) were compared in elicited roots, non-elicited roots and glyphosate-treated elicited roots at different concentrations of glyphosate. It was observed that glyphosate treatment, in addition to 25% suppressions of phenylalanine ammonia-lyase and C2 chain-cleavage enzyme activities as compared to elicited one, also resulted in around 40% suppression of MBALD accumulation, when used in conjunction with yeast extract treatment; in contrast, shikimic acid content was increased as compared to glyphosate untreated ones. These findings suggest that shikimate pathway plays an important role in modulating MBALD biosynthesis in H. indicus roots.

Phenylalanine ammonia-lyase-mediated biosynthesis of 2-hydroxy-4-methoxybenzaldehyde in roots of Hemidesmus indicus.

The fragrant rootstocks of Hemidesmus indicus are known to accumulate 2-hydroxy-4-methoxybenzaldehyde (MBALD), yet, the enzymatic route to this hydroxybenzoate is not known. Therefore, root organs of H. indicus hold promises to unravel the biosynthesis related to this phenolic fragrance. Chitosan treatment at 200mg/L concentration to the excised roots effectively increased phenolic accumulation in both the cortex and cork tissues reaching a peak after 24h treatment and decreasing thereafter. The activity of phenylalanine ammonia-lyase (PAL) enzyme in excised roots also increased upon chitosan elicitation, and the maximum specific activity was recorded after 12h of elicitation. Suppression of PAL in vivo by using a specific irreversible inhibitor aminooxyacetic acid (AOAA) resulted in the decrease in MBALD content, indicating its formation via phenylpropanoid pathway.

Potential of Hemidesmus indicus for phytoextraction of lead from industrially contaminated soils.

Many sites in the industrial region of Kattedan near Hyderabad, Andhra Pradesh (AP), India are contaminated with high concentrations of lead. The use of plants to remove toxic metals from soils (phytoremediation) is emerging as a potential strategy for cost-effective and environmentally sound remediation of contaminated soils. We studied remediation of soils contaminated with lead using a lead hyperaccumulating plant, Hemidesmus indicus. The ability of this plant to accumulate lead in shoots and roots was studied with pot experiments. The results showed that accumulation was maximum in roots for the first 1-3 weeks and later for a contact period of three months, the accumulation rate was maximum in shoots. In addition we used various chelating agents such as EDTA, HEDTA, DTPA and CDTA to determine the best chemical modifier for efficient lead removal from contaminated soils. The effect on lead accumulation of plant in the presence of various metal co-ions was also studied. An attempt was made for the decontamination of lead from five different "Real-life" soils of Kattedan using H. indicus.