Phytochemical profiling, human insulin stability and alpha glucosidase inhibition of Gymnema latifolium leaves aqueous extract: Exploring through experimental and in silico approach.

Diabetes mellitus Type 2 (DM2T) is a rapidly expanding metabolic endocrine disorder worldwide. It is caused due to inadequate insulin secretion by pancreatic beta cells as well as development of insulin resistance. This study aimed to investigate the anti-α-glucosidase, insulin stabilization effect, and non-cytotoxic nature of Gymnema latifolium leaf aqueous extract (GLAE). FTIR analysis revealed the functional groups of compounds present in GLAE. Through LC/ESI-MS/MS analysis, about 12 compounds which belongs to different classes, triterpene glycosides, flavonoids, phenolics, stilbene glycosides and chlorophenolic glycosides were identified. GLAE showed in vitro antioxidant activity. GLAE stabilized insulin by increasing its α-helical content. GLAE inhibited the mammalian α-glucosidase (IC50 = 144 µg/mL) activity through competitive mode (Ki = 61.30 µg/mL). GLAE did not affect the viability of normal cell line (Vero cell line) which shows its non-toxic nature. Molecular docking of phytocompounds identified in GLAE was done with human α-glucosidase and insulin. The top 2 compounds [Gymnema saponin V (GSV) and quercetin 3-(2-galloylglucoside) (QGG) with α-glucosidase; GSV and Z)-resveratrol 3,4'-diglucoside (RDG) with human insulin] with low binding free energy were subjected to 100 ns molecular dynamics simulation to ascertain the stable binding of ligand with protein. The MM/GBSA analysis revealed binding free energy of GSV/α-glucosidase and QGG /α-glucosidase to be - 20.9935 and, - 30.9461 kcal/mol, respectively. Altogether GLAE is valuable source of anti-α-glucosidase inhibitors and insulin stabilizing compounds, suggesting potential lead for further exploration as complementary medicine against DM2T.

Lawsonia inermis flower aqueous extract expressed better anti-alpha-glucosidase and anti-acetylcholinesterase activity and their molecular dynamics.

Lawsonia inermis (henna) has been used in traditional medicine throughout the world and biological property of its flower has been least explored. In the present study, the phytochemical characterization and biological activity (in vitro radical scavenging activity, anti-alpha glucosidase and anti-acetylcholinesterase) of aqueous extract prepared from henna flower (HFAE) was carried out by both Qualitative and quantitative phytochemical analysis and Fourier-transform infrared spectroscopy revealed the functional group of the phytoconstituents such as phenolics, flavonoids, saponin, tannins and glycosides. The phytochemicals present in HFAE was preliminary identified by liquid chromatography/electrospray ionization tandem mass spectrometry. The HFAE showed potent in vitro antioxidant activity and the HFAE inhibited mammalian α-glucosidase (IC50 = 129.1 ± 5.3 µg/ml; Ki = 38.92 µg/ml) and acetylcholinesterase (AChE; IC50 = 137.77 ± 3.5 µg/ml; Ki = 35.71 µg/ml) activity by competitive manner. In silico molecular docking analysis revealed the interaction of active constituents identified in HFAE with human α-glucosidase and AChE. Molecular dynamics simulation for 100 ns showed the stable binding of top two ligand/enzyme complexes with lowest binding energy such as 1,2,3,6-Tetrakis-O-galloyl-beta-D-glucose (TGBG)/human α-glucosidase, Kaempferol 3-glucoside-7-rhamnoside (KGR)/α-glucosidase, agrimonolide 6-O-ß-D-glucopyranoside (AMLG)/human AChE and KGR/AChE. Through MM/GBSA analysis, the binding energy for TGBG/human α-glucosidase, KGR/α-glucosidase, AMLG/human AChE and KGR/AChE was found to be -46.3216, -28.5772, -45.0077 and -47.0956 kcal/mol, respectively. Altogether, HFAE showed an excellent antioxidant, anti-alpha glucosidase and anti-AChE activity under in vitro. This study suggest HFAE with remarkable biological activities could be further explored for therapeutics against type 2 diabetes and diabetes-associated cognitive decline.Communicated by Ramaswamy H. Sarma.

Bio-efficacy of insecticidal molecule emodin against dengue, filariasis, and malaria vectors.

Emodin, a compound isolated from Aspergillus terreus, was studied using chromatographic and spectroscopic methods and compound purity (96%) was assessed by TLC. Furthermore, high larvicidal activity against Aedes aegypti-AeA (LC50 6.156 and LC90 12.450 mg/L), Culex quinquefasciatus-CuQ (8.216 and 14.816 mg/L), and Anopheles stephensi-AnS larvae (6.895 and 15.24 mg/L) was recorded. The first isolated fraction (emodin) showed higher pupicidal activity against AeA (15.449 and 20.752 mg/L). Most emodin-treated larvae (ETL) showed variations in acetylcholine esterase, α and ß-carboxylesterases, and phosphatase activities in the 4th instar, indicating the intrinsic differences in their biochemical changes. ETL had numerous altered tissues, including muscle, gastric caeca, hindgut, midgut, nerve ganglia, and midgut epithelium. Acute toxicity of emodin on brine shrimp Artemia nauplii (54.0 and 84.5 mg/L) and the zebrafish Danio rerio (less toxicity observed) was recorded. In docking studies, Emodin interacted well with odorant-binding-proteins of AeA, AnS, and CuQ with docking scores of - 8.89, - 6.53, and - 8.09 kcal mol-1, respectively. Therefore, A. terreus is likely to be effective against mosquito larvicides.

Characterization of NiONPs synthesized by aqueous extract of orange fruit waste and assessed their antimicrobial and antioxidant potential.

This research was performed to evaluate the nickel oxide nanoparticles (NiONPs) fabricating potential of orange fruit waste (OFW) aqueous extract. Moreover characterize the synthesized OFW-NiONPs through standard techniques such as UV-vis. spectrophotometer, Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and Scanning Electron Microscope (SEM) analyses. Furthermore, the antimicrobial and antioxidant potential of OFW-NiONPs were studied against most common microbial pathogens (Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, and Aspergillus niger) and free radicals (2,2-diphenyl-1-picrylhydrazyl (DPPH), H2O2, OH, and FRAP). A sharp absorbance peak was obtained at 324 nm under UV-vis spectrum analysis that confirmed that the synthesis of OFW-NiONPs and it has been capped and stabilized by numbers of active functional groups studied through FTIR analysis. SEM and DLS analyses revealed that the cubic and triangle shaped OFW-NiONPs with the size intensity distribution was ranging from 21 nm to 130 nm. Interestingly, the OFW-NiONPs showed remarkable antimicrobial activity against the common microbial pathogens in the order of E. coli > A. niger > K. pneumoniae > B. subtilis > S. aureus at increased concentration of 200 µg mL-1. Similarly, the synthesized OFW-NiONPs also possess significant free radicals scavenging activity against DPPH, OH, and FRAP. These results conclude that this OFW-NiONPs can be considered for some biomedical applications after the investigations of some in-vivo research.

Silver nanoparticles loaded graphene-poly-vinylpyrrolidone composites as an effective recyclable antimicrobial agent.

Silver nanoparticles (AgNPs) are often used as antibacterial agents. Here, graphene-silver nanoparticles (G-Ag) and graphene-silver nanoparticles poly-vinylpyrrolidone (G-AgPVPy) were prepared by chemical reduction and in-situ polymerization of vinylpyrrolidone (VPy). The prepared G-Ag and G-AgPVPy composites were characterized using various techniques. The size of the AgNPs on the graphene surface in the prepared G-Ag and G-AgPVPy composites was measured as ∼20 nm. The graphene sheets size in the G-Ag and G-AgPVPy composites were measured as 6.0-2.0 µm and 4.0-0.10 µm, respectively, which are much smaller than graphene sheets in graphite powder (GP) (10.0-3.0 µm). The physicochemical analysis confirmed the formation of G-Ag and G-AgPVPy composites and even the distribution of AgNPs and PVPy on the graphene sheets. The synthesized composites (G-AgPVPy, G-Ag) exhibited a broad-spectrum antibacterial potential against both Gram-negative and Gram-positive bacteria. The lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were calculated as >40 µg/mL using G-Ag and GP, while G-AgPVPy showed as 10 µg/mL against Staphylococcus aureus. Among GP, G-Ag, and G-AgPVPy, G-AgPVPy disturbs the cell permeability, damages the cell walls, and causes cell death efficiently. Also, G-AgPVPy was delivered as a significant reusable antibacterial potential candidate. The MIC value (10 µg/mL) did not change up to six subsequent MIC analysis cycles.

Larvicidal and histopathology effect of endophytic fungal extracts of Aspergillus tamarii against Aedes aegypti and Culex quinquefasciatus.

BACKGROUND: Mosquitoes biolarvicides remain the most important method for mosquito control. The previous studies have shown Aspergillus sp.-expressed larvicidal properties against mosquito species. The present study evaluated larvicidal and histopathological effect of an endophytic fungus Aspergillus tamarii isolated from theCactus stem (Opuntia ficus-indica Mill). METHOD: The molecular identification of isolated A. tamarii was done by PCR amplification (5.8s rDNA) using a universal primer (ITS-1 and ITS-2). The secondary metabolites of A. tamarii was tested for larvicidal activity against Aedes aegypti and Culex quinquefasciatus. Larvicidal bioassay of different concentrations (- 100, 300, 500, 800 and 1000 µg/mL) isolated extracts were done according to the modified protocol. Each test included a set of control groups (i.e. DMSO and distilled water). The lethal concentrations (LC50 and LC90) were calculated by probit analysis. Experimental monitoring duration was 48 h. RESULTS: The ethyl acetate extract from A. tamarii fungus resulted - excellent mosquitocidal effect against Ae. aegypti and Cx. quinquefasciatus mosquitoes, with least LC50 and LC90 values. -After 48 h, the Ae. aegypti expressed better results (LC50 = 29.10, 18.69, 16.76, 36.78 µg/mL and the LC90 = 45.59, 27.66, 27.50, 54.00 µg/mL) followed by Cx. quinquefaciatus (LC50 = 3.23, 24.99, 11.24, 10.95 µg/mL and the LC90 = 8.37, 8.29, 21.36, 20.28 µg/mL). The biochemical level of A. tamarii mycelium extract on both larvae was measured and the results shown a dose dependent activity on the level of AchE, α- and ß-carboxylesterase assay. Gas Chromatography and Mass Spectroscopy (GC-MS) profile of A. tamarii extract reflected three compounds i.e. preg-4-en-3-one, 17. α-hydroxy-17. ß-cyano- (7.39%), trans-3-undecene-1,5-diyne (45.77%) and pentane, 1,1,1,5-tetrachloro- (32.16%) which which might had attributed to larvae mortality. CONCLUSION: The findings of - present study shows that the use of endophytic A. tamarii fungal metabolites for control of dengue and filariasis vectors is promising and needs a semifield and small scale filed trials.

Pouteria campechiana leaf extract and its bioactive compound myricitrin are mosquitocidal against Aedes aegypti and Culex quinquefasciatus

Objective: To test the mosquitocidal potential of leaf extracts of Pouteria campechiana prepared with different solvents and elucidate the structure of an isolated mosquitocidal compound. Methods: The leaf extracts of Pouteria campechiana prepared with three solvents (petroleum benzene, ethyl acetate and acetone) and potential bioactive fractions were tested against various stages of Aedes aegypti and Culex quinquefasciatus by using the WHO protocols, and the chemical profile and its functional groups were identified by GC-MS and Fourier transmission-infrared spectroscopy (FT-IR). The structure of bioactive compound was characterized by nuclear magnetic resonance (NMR) spectral technique. Results: The preliminary phytochemical results revealed the presence of alkaloids, amino acids, flavonoids, quinones, saponins, steroids, tannins, and terpenoids in the acetone extract. A significant toxic potential was observed in the acetone extract against both Aedes aegypti and Culex quinquefasciatus mosquitoes. The acetone extract exhibits remarkable larvicidal (LC

Cloning and Expression of the Organophosphate Pesticide-Degrading α-ß Hydrolase Gene in Plasmid pMK-07 to Confer Cross-Resistance to Antibiotics.

Pesticide residual persistence in agriculture soil selectively increases the pesticide-degrading population and transfers the pesticide-degrading gene to other populations, leading to cross-resistance to a wide range of antibiotics. The enzymes that degrade pesticides can also catabolize the antibiotics by inducing changes in the gene or protein structure through induced mutations. The present work focuses on the pesticide-degrading bacteria isolated from an agricultural field that develop cross-resistance to antibiotics. This cross-resistance is developed through catabolic gene clusters present in an extrachromosomal plasmid. A larger plasmid (236.7 Kbp) isolated from Bacillus sp. was sequenced by next-generation sequencing, and important features such as α-ß hydrolase, DNA topoisomerase, DNA polymerase III subunit beta, reverse transcriptase, plasmid replication rep X, recombination U, transposase, and S-formylglutathione hydrolase were found in this plasmid. Among these, the α-ß hydrolase enzyme is known for the degradation of organophosphate pesticides. The cloning and expression of the α-ß hydrolase gene imply nonspecific cleavage of antibiotics through a cross-resistance phenomenon in the host. The docking of α-ß hydrolase with a spectrum of antibiotics showed a high G-score against chloramphenicol (-3.793), streptomycin (-2.865), cefotaxime (-5.885), ampicillin (-4.316), and tetracycline (-3.972). This study concludes that continuous exposure to pesticide residues may lead to the emergence of multidrug-resistant strains among the wild microbial flora.

Pesticide degrading natural multidrug resistance bacterial flora.

Multidrug-resistant (MDR) bacteria are a growing threat to humans across the world. Antibiotic resistance is a global problem that has developed through continuous antibiotic use, combinatorial antibiotic use, pesticide-antibiotic cross-resistance, and horizontal gene transfer, as well as various other modes. Pesticide-antibiotic cross-resistance and the subsequent expansion of drug-resistant bacteria are critically documented in this review, the primary focus of which is to assess the impact of indiscriminate pesticide use on the development of microbial communities with parallel pesticide and multidrug resistance. The consumption of pesticide-contaminated food products and the use of broad-spectrum antibiotics by humans and in livestock animals have favored the development of both antibiotic and pesticide-resistant bacterial flora via natural selection. Pesticide resistance mainly develops through defensive bacterial adaptations such as biofilm formation, induced mutations, and horizontal/vertical gene transfer through plasmids or transposons, as well as through the increased expression of certain hydrolytic enzymes. Pesticide resistance genes are always transferred as gene clusters, and they may also carry genes essential for antibiotic resistance. Moreover, for some induced mutations, the mutated active site of the affected enzyme may allow degradation of both pesticides and antibiotics, resulting in cross-resistance. A few studies have shown that the sub-lethal exposure of wild-type strains to herbicides induces antibiotic resistance. This review concludes that xenobiotic exposure leads to cross-resistance in wild microbial flora, which requires further study to develop therapeutic approaches to overcome the threats of MDR bacteria and superbugs.

Emergence of multi drug resistance among soil bacteria exposing to insecticides.

Impacts of pesticide exposure on the soil microbial flora and cross resistance to antibiotics have not been well documented. Development of antibiotic resistance is a common issue among soil bacteria which are exposing to pesticides continuously at sub-lethal concentration. The present study was focused to evaluate the correlation between pesticide exposures and evolution of multi drug resistance among isolates collected from soil applied with insecticides. Twenty five insecticide (Monochrotophos) degrading bacteria were isolated from contaminated agricultural soil. The bacterial isolates Bacillus Sps, Bacillus cereus, Bacillus firmus and Bacillus thuringiensis were found to be resistant against chloramphenical, monochrotophos, ampicillin, cefotaxime, streptomycin and tetracycline antibiotics used. Involvement of plasmid in drug as well as insecticide resistant was confirmed through plasmid curing among selected bacterial strains. Bacillus Sps (MK-07), Bacillus cereus (MK-11), Bacillus firmus (MK-13) and Bacillus thuringiensis (MK-24) lost their resistant against insecticides and antibiotics once after removal of plasmid by exposing to 2% sodium dodecyl sulphate. The plasmid was transformed back to bacteria which produced similar derivatives when cultured in Minimal Salt medium (pH 7.0) supplemented with 0.4% of insecticide. Homology modeling was used to prove that organophosphorus hydrolase and able to metabolize all the antibiotics showed positive interaction with high docking score. The present study revealed that persistent of insecticides in the agricultural soil may lead to increasing development of multidrug resistance among soil bacteria.