Antimicrobial resistance in urban river ecosystems.

Antimicrobial resistance (AMR) with the ability to thwart clinical therapies and escalate mortality rates is emerging as one of the most pressing global health and environmental concerns. Urban rivers as an important subsystem of the environment offer galore of ecological services which benefit the city dwellers. However, with increased urbanization, industrialization, and heavy discharge of anthropogenic waste harboring antibiotics, heavy metals, pesticides, antibiotic resistance genes (ARGs), antibiotic resistant bacteria (ARB), urban rivers are becoming major reservoirs of ARGs and a hotspot for accelerated selection of ARB. These ARGs in urban rivers have the potential of being transferred to clinically important pathogens. In addition, urban rivers also act as important vectors for AMR spread. This is mainly due to the direct exposure of humans and animals to the heavily contaminated river water and high mobility of organisms (aquatic animals, pathogenic, non-pathogenic bacteria) as well as the genetic elements including ARGs and mobile genetic elements (MGEs) in the river. However, in spite of recent advocacy for comprehensive research programs aimed to investigate the occurrence, extent and major drivers of AMR in urban rivers globally, such studies are missing largely. This review encompasses the issues of AMR, major drivers and their vital roles in the evolution and spread of ARB with an emphasis on sources and hotspots of diverse ARGs in urban rivers contributing to co-occurrence of ARGs and MGEs. Further, the causal factors leading to adverse effects of antibiotic-load to river organisms with an elaboration on the current measures to eradicate the ARB, ARGs, and remove antibiotics from the urban river ecosystems are also discussed. A perspective review of current and emerging strategies with potentials of combating AMR in urban river ecosystems including advanced water treatment methodologies and floating islands or constructed wetlands.

Microplastic-associated pathogens and antimicrobial resistance in environment.

The ubiquitous use of microplastics and their release into the environment especially the water bodies by anthropogenic/industrial activities are the major resources for microplastic contamination. The widespread and often injudicious use of antimicrobial drugs or antibiotics in various sectors including human health and hygiene, agriculture, animal husbandry and food industries are leading to the release of antibiotics into the wastewater/sewage and other water bodies, particularly in urban setups and thus leads to the antimicrobial resistance (AMR) in the microbes. Microplastics are emerging as the hubs as well as effective carriers of these microbial pathogens beside their AMR-genes (ARGs) in marine, freshwater, sewage/wastewater, and urban river ecosystems. These drug resistant bacteria interact with microplastics forming synthetic plastispheres, the ideal niche for biofilm formations which in turn facilitates the transfer of ARGs via horizontal gene transfer and further escalates the occurrence and levels of AMR. Microplastic-associated AMR is an emerging threat for human health and healthcare besides being a challenge for the research community for effective management/address of this menace. In this review, we encompass the increasing prevalence of microplastics in environment, emphasizing mainly on water environments, how they act as centers and vectors of microbial pathogens with their associated bacterial assemblage compositions and ultimately lead to AMR. It further discusses the mechanistic insights on how microplastics act as hosts of biofilms (creating the plastisphere). We have also presented the modern toolbox used for microplastic-biofilm analyses. A review on potential strategies for addressing microplastic-associated AMR is given with recent success stories, challenges and future prospects.

Combating Drug-Resistant Bacteria Using Photothermally Active Nanomaterials: A Perspective Review.

Injudicious use of antibiotics has been the main driver of severe bacterial non-susceptibility to commonly available antibiotics (known as drug resistance or antimicrobial resistance), a global threat to human health and healthcare. There is an increase in the incidence and levels of resistance to antibacterial drugs not only in nosocomial settings but also in community ones. The drying pipeline of new and effective antibiotics has further worsened the situation and is leading to a potentially "post-antibiotic era." This requires novel and effective therapies and therapeutic agents for combating drug-resistant pathogenic microbes. Nanomaterials are emerging as potent antimicrobial agents with both bactericidal and potentiating effects reported against drug-resistant microbes. Among them, the photothermally active nanomaterials (PANs) are gaining attention for their broad-spectrum antibacterial potencies driven mainly by the photothermal effect, which is characterized by the conversion of absorbed photon energy into heat energy by the PANs. The current review capitalizes on the importance of using PANs as an effective approach for overcoming bacterial resistance to drugs. Various PANs leveraging broad-spectrum therapeutic antibacterial (both bactericidal and synergistic) potentials against drug-resistant pathogens have been discussed. The review also provides deeper mechanistic insights into the mechanisms of the action of PANs against a variety of drug-resistant pathogens with a critical evaluation of efflux pumps, cell membrane permeability, biofilm, and quorum sensing inhibition. We also discuss the use of PANs as drug carriers. This review also discusses possible cytotoxicities related to the therapeutic use of PANs and effective strategies to overcome this. Recent developments, success stories, challenges, and prospects are also presented.

Embelin-loaded chitosan gold nanoparticles interact synergistically with ciprofloxacin by inhibiting efflux pumps in multidrug-resistant Pseudomonas aeruginosa and Escherichia coli.

A global upsurge in emergence and spread of antibiotic resistance (ABR) in bacterial populations is a serious threat for human health. Unfortunately, ABR is no longer confined to nosocomial environments and is frequently reported from community microbes as well. The ABR is resulting in shrinking potent antibiotics pool and thus necessitating novel and alternative therapies and therapeutics. Current investigation was aimed to assess the synergistic potential of a synthesized, phytomolecule-loaded, polysaccharide-stabilized metallic nanoparticles (NPs) against Pseudomonas aeruginosa (PA) and Escherichia coli (EC) isolated from river waters. ABR profiling of these strains characterized them as multidrug resistant (MDR). Synthesized embelin (Emb, isolated from Embelia tsjeriam-cottam)-loaded, chitosan-gold (Emb-Chi-Au) NPs were assessed for their potential synergistic activity with ciprofloxacin (CIP) via checker-board assay and time-kill curve analysis. The NPs reduced the minimal inhibitory concentration (MIC) of CIP by 16- and 4-fold against MDR PA (PA-r) and EC (EC-r) strains, respectively. Fractional inhibitory concentration (FIC) indices with ≤0.5 values confirmed the synergy between the Emb-Chi-Au NPs and CIP, which was further confirmed at ½ MICs in both PA-r and EC-r via time-kill curve analysis. In order to decipher the mode of action, efflux pump inhibitory effects of Emb-Chi-Au NPs were evaluated in terms of the increase in the EtBr mediated fluorescence in control versus NP-treated MDR strains. Molecular docking based in silico simulations were used to predict the interactions between Emb and the active sites of the efflux pump related proteins in PA-r (MexA, MexB and OprM) and EC-r (AcrA, AcrB and TolC), which revealed the probable bond formation between Emb and respective amino acid residues.

Plant synthetic biology for producing potent phyto-antimicrobials to combat antimicrobial resistance.

Inappropriate and injudicious use of antimicrobial drugs in human health, hygiene, agriculture, animal husbandry and food industries has contributed significantly to rapid emergence and persistence of antimicrobial resistance (AMR), one of the serious global public health threats. The crisis of AMR versus slower discovery of newer antibiotics put forth a daunting task to control these drug-resistant superbugs. Several phyto-antimicrobials have been identified in recent years with direct-killing (bactericidal) and/or drug-resistance reversal (re-sensitization of AMR phenotypes) potencies. Phyto-antimicrobials may hold the key in combating AMR owing to their abilities to target major microbial drug-resistance determinants including cell membrane, drug-efflux pumps, cell communication and biofilms. However, limited distribution, low intracellular concentrations, eco-geographical variations, beside other considerations like dynamic environments, climate change and over-exploitation of plant-resources are major blockades in full potential exploration phyto-antimicrobials. Synthetic biology (SynBio) strategies integrating metabolic engineering, RNA-interference, genome editing/engineering and/or systems biology approaches using plant chassis (as engineerable platforms) offer prospective tools for production of phyto-antimicrobials. With expanding SynBio toolkit, successful attempts towards introduction of entire gene cluster, reconstituting the metabolic pathway or transferring an entire metabolic (or synthetic) pathway into heterologous plant systems highlight the potential of this field. Through this perspective review, we are presenting herein the current situation and options for addressing AMR, emphasizing on the significance of phyto-antimicrobials in this apparently post-antibiotic era, and effective use of plant chassis for phyto-antimicrobial production at industrial scales along with major SynBio tools and useful databases. Current knowledge, recent success stories, associated challenges and prospects of translational success are also discussed.

Biotic elicitors enhance diosgenin production in Helicteres isora L. suspension cultures via up-regulation of CAS and HMGR genes.

In an attempt to find an alternative and potent source of diosgenin, a steroidal saponin in great demand for its pharmaceutical importance, Helicteres isora suspension cultures were explored for diosgenin extraction. The effect of biotic elicitors on the biosynthesis of diosgenin, in suspension cultures of H. isora was studied. Bacterial as well as fungal elicitors such as Escherichia coli, Bacillus subtilis, Saccharomyces cerevisiae and Aspergillus niger were applied at varying concentrations to investigate their effects on diosgenin content. The HPLC based quantification of the treated samples proved that amongst the biotic elicitors, E. coli (1.5%) proved best with a 9.1-fold increase in diosgenin content over respective control cultures. Further, the scaling-up of the suspension culture to shake-flask and ultimately to bioreactor level were carried out for production of diosgenin. During all the scaling-up stages, diosgenin yield obtained was in the range between 7.91 and 8.64 mg l-1, where diosgenin content was increased with volume of the medium. The quantitative real-time PCR (qRT-PCR) analysis showed biotic elicitors induced the expression levels of regulatory genes in diosgenin biosynthetic pathway, the 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and cycloartenol synthase (CAS), which can be positively correlated with elicited diosgenin contents in those cultures. The study holds significance as H. isora represents a cleaner and easy source of diosgenin where unlike other traditional sources, it is not admixed with other steroidal saponins, and the scaled-up levels of diosgenin achieved herein have the potential to be explored commercially.

Phytochemical profile, anti-oxidant, anti-inflammatory, and anti-proliferative activities of Pogostemon deccanensis essential oils.

Essential oils (EOs) obtained from aerial parts of Pogostemon deccanensis were analyzed for GC-MS profiling, and evaluated for antioxidant, anti-inflammatory, and anti-proliferative activities. GC-MS analysis revealed a total of 47 constituents, establishing the EOs rich in sesquiterpene with > 20 sesquiterpenes constituting around 77% of the total EO yield. Major constituents included Curzerene (Benzofuran, 6-ethenyl-4,5,6,7-tetrahydro-3,6-dimethyl-5-isopropenyl-, trans-) (26.39%) and epi-Cadinol (22.68%), Ethanone, 1-(2,4,6-trihydroxyphenyl) (6.83%, Acetophenones), and Boldenone (3.47%, anabolic steroid). EOs found to be rich in phytochemicals attributed for antioxidant potentials of aromatic/medicinal plants, viz., flavonoids (2.71 µg quercetin equivalents g-1 EO), total phenols (3.94 µg gallic acid equivalents (GAE) g-1 EO), carotenoids (14.3 µg ß-carotene equivalents g-1 EO), and ascorbic acid (2.21 µg ascorbic acid equivalents g-1 EO). P. deccanensis EOs exhibited striking antioxidant activities assessed by wide range of assays including ferric reducing antioxidant potential (FRAP, 255.3 GAE at 2 µg mL-1 EO), total antioxidant activity (TAA, 264.3 GAE at 2 µg ml-1) of EO, DPPH (65% inhibition at 2 µg mL-1), and OH (58% inhibition at 2 µg mL-1) scavenging. Interestingly, EOs showed considerably higher anti-lipid peroxidation activity than the standard antioxidant molecule ascorbic acid, with 50% protection by 1.29 µg mL-1 EO against 20.0 µg mL-1 standard. EOs showed strong anti-inflammatory activity with 50% inhibition at 1.95 µg mL-1 EO. The anti-proliferative activity of EOs was tested against mouse cancer cell line and the EOs proved a potent anti-proliferative agent with only 2.1% cell survival at 2 µg mL-1 EO, whereas the EOs were largely non-toxic-to-normal (non-cancerous) cells with approximately 80% cell survival at the 2 µg mL-1 EOs. This being the first attempt of phytochemical profiling and wide array of biological activities of P. deccanensis EOs holds significance as the striking activities were observed at very low concentrations, in some cases at lower than the commercial standards, and has, therefore, great potential for pharmaceutical or commercial exploration.

Plant small RNAs: the essential epigenetic regulators of gene expression for salt-stress responses and tolerance.

Saline environment cues distort the plant growth, development and crop yield. Epigenetics has emerged as one of the prime themes in plant functional genomics for molecular-stress-physiology research, as copious studies have provided new visions into the epigenetic control of stress adaptations. The epigenetic control is associated with the regulation of the expression of stress-related genes which also comprises many steady alterations inherited in next cellular generation as stress memory. These epigenetic amendments also implicate induction of small RNA (sRNA)-mediated fine-tuning of transcriptional and post-transcriptional regulations of gene expression. These tiny (19-24 nt) RNA species, particularly microRNAs (miRNAs) besides endogenous small interfering RNA (siRNA) have emerged as important responsive entities for epigenetic modulation of salt-stress effects on plants. There is a recent upsurge in development of tools and databases useful for prediction, identification and validation of small RNAs (sRNAs) and their target messenger RNAs (mRNAs). Therefore, these small but key regulatory molecules have received a wide attention in post-genomic era as potential targets for engineering stress tolerance in major glycophytic crops, though it is yet to be explored optimally. This review aims to provide critical updates on plant sRNAs as key epigenetic regulators of plant salt-stress responses, their target prediction and validation, computational tools and databases available for plant small RNAs, besides discussing their roles in salt-stress regulatory networks and adaptive mechanisms in plants, with special emphasis on their exploration for engineering salinity tolerance in plants.

Inhibiting Bacterial Drug Efflux Pumps via Phyto-Therapeutics to Combat Threatening Antimicrobial Resistance.

Antibiotics, once considered the lifeline for treating bacterial infections, are under threat due to the emergence of threatening antimicrobial resistance (AMR). These drug-resistant microbes (or superbugs) are non-responsive to most of the commonly used antibiotics leaving us with few treatment options and escalating mortality-rates and treatment costs. The problem is further aggravated by the drying-pipeline of new and potent antibiotics effective particularly against the drug-resistant strains. Multidrug efflux pumps (EPs) are established as principal determinants of AMR, extruding multiple antibiotics out of the cell, mostly in non-specific manner and have therefore emerged as potent drug-targets for combating AMR. Plants being the reservoir of bioactive compounds can serve as a source of potent EP inhibitors (EPIs). The phyto-therapeutics with noteworthy drug-resistance-reversal or re-sensitizing activities may prove significant for reviving the otherwise fading antibiotics arsenal and making this combination-therapy effective. Contemporary attempts to potentiate the antibiotics with plant extracts and pure phytomolecules have gained momentum though with relatively less success against Gram-negative bacteria. Plant-based EPIs hold promise as potent drug-leads to combat the EPI-mediated AMR. This review presents an account of major bacterial multidrug EPs, their roles in imparting AMR, effective strategies for inhibiting drug EPs with phytomolecules, and current account of research on developing novel and potent plant-based EPIs for reversing their AMR characteristics. Recent developments including emergence of in silico tools, major success stories, challenges and future prospects are also discussed.

MicroRNAs As Potential Targets for Abiotic Stress Tolerance in Plants.

The microRNAs (miRNAs) are small (20-24 nt) sized, non-coding, single stranded riboregulator RNAs abundant in higher organisms. Recent findings have established that plants assign miRNAs as critical post-transcriptional regulators of gene expression in sequence-specific manner to respond to numerous abiotic stresses they face during their growth cycle. These small RNAs regulate gene expression via translational inhibition. Usually, stress induced miRNAs downregulate their target mRNAs, whereas, their downregulation leads to accumulation and function of positive regulators. In the past decade, investigations were mainly aimed to identify plant miRNAs, responsive to individual or multiple environmental factors, profiling their expression patterns and recognizing their roles in stress responses and tolerance. Altered expressions of miRNAs implicated in plant growth and development have been reported in several plant species subjected to abiotic stress conditions such as drought, salinity, extreme temperatures, nutrient deprivation, and heavy metals. These findings indicate that miRNAs may hold the key as potential targets for genetic manipulations to engineer abiotic stress tolerance in crop plants. This review is aimed to provide recent updates on plant miRNAs, their biogenesis and functions, target prediction and identification, computational tools and databases available for plant miRNAs, and their roles in abiotic stress-responses and adaptive mechanisms in major crop plants. Besides, the recent case studies for overexpressing the selected miRNAs for miRNA-mediated enhanced abiotic stress tolerance of transgenic plants have been discussed.

Antimicrobial potentials of Helicteres isora silver nanoparticles against extensively drug-resistant (XDR) clinical isolates of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a leading opportunistic pathogen and its expanding drug resistance is a growing menace to public health. Its ubiquitous nature and multiple resistance mechanisms make it a difficult target for antimicrobial chemotherapy and require a fresh approach for developing new antimicrobial agents against it. The broad-spectrum antibacterial effects of silver nanoparticles (SNPs) make them an excellent candidate for use in the medical field. However, attempts made to check their potency against extensively drug-resistant (XDR) microbes are meager. This study describes the biosynthesis and biostabilization of SNPs by Helicteres isora aqueous fruit extract and their characterization by ultraviolet-visible spectroscopy, transmission electron microscopy, dynamic light scattering, X-ray diffraction, and Fourier transform infrared spectroscopy. Majority of SNPs synthesized were of 8--20-nm size. SNPs exhibited dose-dependent antibacterial activities against four XDR P. aeruginosa (XDR-PA) clinical isolates as revealed by growth curves, with a minimum inhibitory concentration of 300 µg/ml. The SNPs exhibited antimicrobial activity against all strains, with maximum zone of inhibition (16.4 mm) in XRD-PA-2 at 1000 µg/ml. Amongst four strains, their susceptibilities to SNPs were in the following order: XDR-PA-2 > XDR-PA-4 > XDR-PA-3 > XDR-PA-1. The exposure of bacterial cells to 300 µg/ml SNPs resulted into a substantial leakage of reducing sugars and proteins, inactivation of respiratory chain dehydrogenases, and eventual cell death. SNPs also induced lipid peroxidation, a possible underlying factor to membrane porosity. The effects were more pronounced in XDR-PA-2 which may be correlated with its higher susceptibility to SNPs. These results are indicative of SNP-induced turbulence of membranous permeability as an important causal factor in XDR-PA growth inhibition and death.

In vitro regeneration and ploidy level analysis of Eulophia ochreata Lindl.

Various parameters including explant-type, medium compositions, use of phytohormones and additives were optimized for direct and indirect regeneration of E. ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs) proved to be the best explants for shoot initiation, proliferation and callus induction. Murashige and Skoog's (MS) medium containing 2.5 mg L(-1) 6-benzylaminopurine (BAP), 1.0 mg L(-1) kinetin (Kin) and additives (adenine sulfate, arginine, citric acid, 30 mg L(-1) each and 50 mg L(-1) ascorbic acid) was optimal for shoot multiplication (12.1 shoots and 7.1 PLBs per explant with synchronized growth), which also produced callus. Shoot number was further increased with three successive subcultures on same media and approximately 40 shoots per explant were achieved after 3 cycles of 30 days each. Additives and casein hydrolysate (CH) showed advantageous effects on indirect shoot regeneration via protocorm-derived callus. Optimum indirect regeneration was achieved on MS containing additives, 500 mg L(-1) CH, 2.5 mg L(-1) BAP and 1.0 mg L(-1) Kin with 30 PLBs and 6 shoots per callus mass (approximately 5 mm size). The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L(-1) indole-3-butyric acid, 200 mg L(-1) activated charcoal and additives. The rooted plantlets were hardened and transferred to greenhouse with 63% survival rate. Flow-cytometry based DNA content analysis revealed that the ploidy levels were maintained in in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.

Hairy Root Induction in Helicteres isora L. and Production of Diosgenin in Hairy Roots.

Mature seeds of Helicteres isora L. were collected from seven geographical locations of Maharashtra and Goa (India) and evaluated for diosgenin (a bioactive steroidal sapogenin of prime importance) extraction and quantification. Chemotypic variations were evidenced with diosgenin quantity ranging from 33 µg g(-1) seeds (Osmanabad forests) to 138 µg g(-1) (Khopoli region). Nodal and leaf explants from in vitro-raised seedlings were used for callus and Agrobacterium-mediated transformation, respectively. Compact, hard, whitish-green callus (2.65 g explant(-1)) was obtained on MS + 13.32 µM BAP + 2.32 µM Kin after 30 days of inoculation. Various parameters including types of explant and Agrobacterium strain, culture density, duration of infection and various medium compositions were optimized for hairy root production. A. rhizogenes strain ATCC-15834 successfully induced hairy roots from leaf explants (1 cm(2)) with 42 % efficiency. Transgenic status of the roots was confirmed by PCR using rolB and VirD specific primers. Hairy roots showed an ability to synthesize diosgenin. Diosgenin yield was increased ~8 times in hairy roots and ~5 times in callus than the seeds of wild plants. Enhanced diosgenin content was associated with proline accumulation in hairy roots. This is the first report on induction of hairy roots in H. isora.

Antibacterial & antiplasmid activities of Helicteres isora L.

BACKGROUND & OBJECTIVES: The multiple drug resistance (MDR) is a serious health problem and major challenge to the global drug discovery programmes. Most of the genetic determinants that confer resistance to antibiotics are located on R-plasmids in bacteria. The present investigation was undertaken to investigate the ability of organic extract of the fruits of Helicteres isora to cure R-plasmids from certain clinical isolates. METHODS: Active fractions demonstrating antibacterial and antiplasmid activities were isolated from the acetone extracts of shade dried fruits of H. isora by bioassay guided fractionation. Minimal inhibitory concentration (MIC) of antibiotics and organic extracts was determined by agar dilution method. Plasmid curing activity of organic fractions was determined by evaluating the ability of bacterial colonies (pre treated with organic fraction for 18 h) to grow in the presence of antibiotics. The physical loss of plasmid DNA in the cured derivatives was further confirmed by agarose gel electrophoresis. RESULTS: The active fraction did not inhibit the growth of either the clinical isolates or the strains harbouring reference plasmids even at a concentration of 400 microg/ml. However, the same fraction could cure plasmids from Enterococcus faecalis, Escherichia coli, Bacillus cereus and E. coli (RP4) at curing efficiencies of 14, 26, 22 and 2 per cent respectively. The active fraction mediated plasmid curing resulted in the subsequent loss of antibiotic resistance encoded in the plasmids as revealed by antibiotic resistance profile of cured strains. The physical loss of plasmid was also confirmed by agarose gel electrophoresis. INTERPRETATION & CONCLUSIONS: The active fraction of acetone extract of H. isora fruits cured R-plasmids from Gram-positive and Gram-negative clinical isolates as well as reference strains. Such plasmid loss reversed the multiple antibiotic resistance in cured derivatives making them sensitive to low concentrations of antibiotics. Acetone fractions of H. isora may be a source to develop antiplasmid agents of natural origin to contain the development and spread of plasmid borne multiple antibiotic resistance.

Cytotoxic activity of 9,10-dihydro-2,5-dimethoxyphenanthrene-1,7-diol from Eulophia nuda against human cancer cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Eulophia nuda L. (Orchidaceae) is a medicinally important terrestrial orchid used for the treatment of tumours and various health problems by the local healers throughout the Western Ghats region in Maharashtra (India). AIM OF THE STUDY: To isolate the active molecule from Eulophia nuda and to study its cytotoxic potential against human cancer cells. MATERIALS AND METHODS: The crude methanolic extract of Eulophia nuda tubers was fractionated by stepwise gradient of the solvents-chloroform-methanol to isolate the pure compound. Isolated pure compound was assessed for its cytotoxic potential against human breast cancer cell lines, MCF-7 and MDA-MB-231 using MTT assay. Structure elucidation of the isolated active compound was carried out by extensive spectroscopic analysis including (1)H NMR, (13)C NMR, NOESY, COSY, LC-MS and IR. RESULTS: The isolated active molecule was identified as phenanthrene derivative 9,10-dihydro-2,5-dimethoxyphenanthrene-1,7-diol. This compound showed good antiproliferative activity against human breast cancer cell lines MCF-7 (91%) and MDA-MB-231 (85%) at 1000 microg/ml concentration. CONCLUSION: 9,10-Dihydro-2,5-dimethoxyphenanthrene-1,7-diol from Eulophia nuda tubers showed good growth suppressive effect against human cancer cell lines MCF-7 and MDA-MB-231 making it a potential biomolecule against human cancer.

Antiplasmid activity of 1'-acetoxychavicol acetate from Alpinia galanga against multi-drug resistant bacteria.

ETHNOPHARMACOLOGICAL RELEVANCE: Alpinia galanga (L.) Swartz is traditionally used in the treatment of various ailments across India, China, and Southeast Asian countries. In India it is a reputed drug in indigenous system of medicine and largely used as antibacterial and antiseptic. In southern India the rhizomes has been used as a domestic remedy against bacterial infections. AIM OF THE STUDY: To identify a potential antiplasmid compound from Alpinia galanga against multi-drug resistant bacteria. MATERIALS AND METHODS: The crude rhizome extract of Alpinia galanga was prepared in acetone. Antibacterial activity was checked by MIC and antiplasmid activity was checked by SIC. The principal compound responsible for the antiplasmid activity, in the crude extract, was identified by bioassay guided fractionation using hexane-acetone. Antibiotic resistance profile of plasmid harboring strains and plasmid cured strains was determined by disc diffusion method. RESULTS: The crude acetone extract of the rhizomes of Alpinia galanga exhibited antiplasmid activity against Salmonella typhi, Escherichia coli and vancomycin resistant Enterococcus faecalis with an efficiency of 92%, 82% and 8% respectively at 400 microg/ml SIC. The principal compound responsible for the activity was identified as 1'-acetoxychavicol acetate. 1'-Acetoxychavicol acetate demonstrated the ability to cure plasmid encoded antibiotic resistance in various multi-drug resistant bacterial strains of clinical isolates such as Enterococcus faecalis, Salmonella typhi, Pseudomonas aeruginosa, Escherichia coli and Bacillus cereus with curing efficiency of 66%, 75%, 70%, 32% and 6% respectively at SIC of 400-800 microg/ml. CONCLUSION: 1'-Acetoxychavicol acetate mediated R-plasmid curing significantly reduced the minimal inhibitory concentration of antibiotics required to inhibit growth of bacteria, thus making the antibiotic treatment more effective.

A potential plasmid-curing agent, 8-epidiosbulbin E acetate, from Dioscorea bulbifera L. against multidrug-resistant bacteria.

Bioassay-guided fractionation of an aqueous methanolic extract of Dioscorea bulbifera L. bulbs was performed using organic solvents. A novel plasmid-curing compound was identified as 8-epidiosbulbin E acetate (EEA) (norditerpene) on the basis of modern spectroscopic analysis and X-ray crystallography. EEA exhibited broad-spectrum plasmid-curing activity against multidrug-resistant (MDR) bacteria, including vancomycin-resistant enterococci. EEA cured antibiotic resistance plasmids (R-plasmids) from clinical isolates of Enterococcus faecalis, Escherichia coli, Shigella sonnei and Pseudomonas aeruginosa with 12-48% curing efficiency. The reference plasmids of Bacillus subtilis (pUB110), E. coli (RP4), P. aeruginosa (RIP64) and Salmonella typhi (R136) were cured with efficiency ranging from 16% to 64%. EEA-mediated R-plasmid curing decreased the minimal inhibitory concentration of antibiotics against MDR bacteria, thus making antibiotic treatment more effective. The antibiotic resistance pattern revealed that the compound was effective in the reversal of bacterial resistance to various antibiotics. In addition, the compound did not show any cytotoxicity against a broad range of human cancer cell lines, namely MCF-7 (breast cancer), SiHa (cervical cancer) and A431 (epidermal carcinoma), and hence has the potential to be used as a lead compound for drug discovery programmes.