Antibiofilm and quorum sensing inhibitory activity of Achyranthes aspera on cariogenic Streptococcus mutans: an in vitro and in silico study.

CONTEXT: Traditionally, many cultures use chewing sticks for oral hygiene maintenance. When properly used, these chewing sticks are found to be efficient due to the combined effect of mechanical cleaning, enhanced salivation and the antimicrobial action of leached out plant compounds. OBJECTIVE: Achyranthes aspera L. (Amaranthaceae), an ethanomedicinal herb was evaluated for its potential to inhibit growth and biofilm formation by cariogenic isolate Streptococcus mutans as an alternative means of caries management by quorum quenching (QQ). MATERIALS AND METHODS: Biofilm forming cariogenic isolates were isolated and their susceptibility to the petroleum ether, benzene, methanol, aqueous extracts of A. aspera was evaluated. Gas chromatography-mass spectrometry (GC-MS), phytochemical analyses and structure-based virtual screening for their quorum sensing (QS) inhibitory activities, drug-likeness and bioavailability were also carried out. RESULTS: The biofilm inhibition percentage obtained for methanol, benzene, petroleum ether and aqueous extracts (125 µg/mL) were ≤94%, ≤74%, ≤62% ≤42%, respectively. GC-MS analyses indicated 61 compounds, of which betulin recorded efficient interaction exhibiting comparable binding energy of -8.72 with S. mutans glycosyltransferase (GTF-SI) whereas 3,12-oleandione exhibited binding energy -5.92 with OmpR subfamily QS regulatory DNA-binding response regulator. Computer-assisted molecular descriptor and Lipinski's rule violation calculation uncovered the presence of more drug-like compounds. DISCUSSION AND CONCLUSION: Anticaries bioactive compounds of A. aspera with higher QS response regulator binding energy, low toxicity and optimal pharmacokinetic properties were revealed. These compounds with possible QQ ability hold the potential for use as anticaries drug leads and antibiofilm preventative medicine.

Simple and rapid protocol for the isolation of PCR-amplifiable DNA from medicinal plants.

Medicinal plant species has a valuable economic importance because of its usage as pharmaceuticals, nutritional, as well as its use in popular medication. For DNA-based techniques, nanogram quantities of the purified DNA are requisite to amplify and yield sufficient amounts of PCR products. SDS-based DNA isolation method was used to extract DNA from 11 species of different aromatic and medicinal plants collected from Saudi Arabia. Three hundred milligrams of fresh shredded plant material was necessary. The DNA purity was further confirmed by agarose gel, restriction endonuclease digestion and microsatellite primed-polymerase chain reaction (MP-PCR). DNA yields ranged from 10-20 µg (in 100-µL elution volumes) from all plant material evaluated. The DNA obtained was free of any contaminating proteins, polysaccharides and colored pigments. The extracted genomic DNA was found suitable for restriction digestion and PCR amplification. Our experimental procedure provides an easy, suitable, non-toxic, cheap, and quick process for the amplification of DNA from medical plant tissue.

Molecular characterization of the pathogenic plant fungus Rhizoctonia solani (Ceratobasidiaceae) isolated from Egypt based on protein and PCR-RAPD profiles.

Twenty-one isolates of Rhizoctonia solani were categorized into three anastomosis groups consisting of AG-4-HG-I (eight isolates), AG-2-2 (nine isolates) and AG-5 (four isolates). Their pathogenic capacities were tested on cotton cultivar Giza 86. Pre-emergence damping-off varied in response to the different isolates; however, the differences were not significant. Soluble proteins of the fungal isolates were electrophoresed using SDS-PAGE and gel electrophoreses. A dendrogram of the protein banding patterns by the UPGMA of arithmetic means placed the fungal isolates into distinct groups. There was no evidence of a relationship between protein dendrogram, anastomosis grouping or level of virulence or geographic origin. The dendrogram generated from these isolates based on PCR analysis with five RAPD-PCR primers showed high levels of genetic similarity among the isolates from the same geographical locations. There was partially relationship between the genetic similarity and AGs or level of virulence or geographic origin based on RAPD dendrogram. These results demonstrate that RAPD technique is a useful tool in determining the genetic characterization among isolates of R. solani.