Synthesis, characterisation and in vitro anticancer activity of conjugated protease inhibitor-silver nanoparticles (AgNPs-PI) against human breast MCF-7 and prostate PC-3 cancer cell lines.

The conjugated silver nanoparticles using biomolecules have attracted great attention of researchers because physical dimensions and surface chemistry play important roles in toxicity and biocompatibility of AgNPs. Hence, in the current study, synthesis of bio-conjugated AgNPs with protein protease inhibitor (PI) isolated from Streptomyces spp. is reported. UV-visible spectra of PI and AgNPs showed stronger peaks at 280 and 405 nm, confirming the synthesis of conjugated AgNPs-PI. TEM and SEM images of AgNPs-PI showed spherical-shaped nanoparticles with a slight increase in particle size and thin amorphous layer around the surface of silver nanomaterial. Circular dichroism, FT-IR and fluorescence spectral studies confirmed AgNPs-PI conjugation. Conjugated AgNPs-PI showed excellent anticancer potential than AgNPs and protease inhibitor separately on human breast MCF-7 and prostate PC-3 cell lines. The findings revealed that surface modification of AgNPs with protein protease inhibitor stabilised the nanomaterial and increased its anticancer activity.

Inhibitory kinetics and mechanism of pentacyclic triterpenoid from endophytic Colletotrichum gigasporum against pancreatic lipase.

The burden of obesity is increasing all over the world. Except for Orlistat, no effective anti-obesity drug is currently available. Therefore, a search for the new anti-obesity compound is need of time. This study demonstrates macromolecular interaction and inhibitory effect of pentacyclic triterpenoids (PTT) on pancreatic lipase (PL). In the present study PTTs from endophytic Colletotrichum gigasporum were found to show significant inhibitory activity against PL with IC50 of 16.62 ± 1.43 µg/mL. The PTT isolated through bioassay-guided isolation showed a dose-dependent (R2 = 0.915) inhibition against porcine PL and the results were comparable with the standard (Orlistat). Based on inhibition kinetic data, the gradual increase in Km (app) with increasing PTT concentration indicated that the mode of interaction of PTT with PL was a competitive type, and it directly competed with the substrate (pNPB) for the active site of PL. In vivo studies in Wistar rats at the oral dose (100 mg/kg body weight) of PTT significantly decreased (p < 0.05) incremental plasma triglyceride levels as compared to group B and TG absorption was down-regulated up to 49.18% vis a vis group D animals. The isolated PTT was identified as lupeol based on chromatographic and spectral data. The endophytic isolate was identified as Colletotrichum gigasporum based on morphology and ITS gene sequencing. The present study indicated that PTT had the potential to be used as a natural PL inhibitor in the treatment of obesity and the isolated endophyte can be a valuable bioresource for it.

Ethnopharmacology, phytochemistry, and biotechnological advances of family Apocynaceae: A review.

The family Apocynaceae is one of the largest and important families in angiosperm. Several members of this family have medicinal properties and have been in the treatment of various ailments. Most of them are consumed as food by tribal people whereas a few plants are used as source of poison. Members of family Apocynaceae are rich in alkaloids, terpenoids, steroids, flavonoids, glycosides, simple phenols, lactones, and hydrocarbons. Other compounds such as sterols, lignans, sugars, lignans, and lactones have been isolated and systematically studied. Few studies have reported antioxidant, anti-inflammatory, antimicrobial, and cytotoxic activities of crude extracts as well as single compound(s) isolated from various members of the family Apocynaceae. Holarrhena antidysenterica, Rauvolfia serpentina, Carissa carandas, and Tabernaemontana divaricata are the extensively studied plants in this family. The present review provides a detailed outlook on ethnopharmacology, phytochemistry, and biological activities of selected members of this family. Moreover, it also covers the biotechnological advances used for large-scale production of bioactive compounds of therapeutic interest along with plant tissue culture-based approaches for conservation of this medicinally valuable family.

Evaluation of Larvicidal Activity of Essential Oil from Leaves of Coccinia grandis against Three Mosquito Species.

BACKGROUND: To study the chemical constituents and larvicidal activity of essential oil extracted from the leaves of Coccinia grandis against three mosquito species. METHODS: Essential oil was extracted by hydro distillation using clevenger apparatus and was analyzed for chemical constituents by gas chromatography-mass spectrophotometry (GC-MS). Larvicidal activity was recorded after 12 and 24h of post-exposure against three mosquito species, Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus. Dead larvae were identified when they failed to move after probing with a needle in the siphon or cervical region. The LC50 and LC90 values for three mosquito larvae were calculated by Probit analysis. RESULTS: The GC-MS analysis revealed that essential oil contains 23 different constituents. Out of these 23 constituents, major constituents identified were n-tetracosane (39.18%), n-eicosane (30.04%), tetratriacotane (2.97%), 7-octadecanal (2.81%), and tricosane (2.31%). Essential oil from leaves of Coccinia grandis exhibited significant larvicidal activity against An. stephensi with LC50 and LC90 values 39.41ppm and 123.24ppm, respectively. This was followed by Ae. aegypti and Cx. quinquefasciatus with LC50 and LC90 values of 48.20ppm, 131.84ppm and 52.80ppm, 135.48ppm, respectively after 24h of exposure. CONCLUSION: The results could be useful in developing a cost effective, ecofriendly, region specific and practical strategy for the control of mosquito vectors.

Experimental Validation of Antidiabetic Potential of Cayratia trifolia (L.) Domin: An Indigenous Medicinal Plant.

The present study was undertaken to evaluate antidiabetic and antioxidant activities of Cayratia trifolia root extract against streptozotocin induced diabetes in experimental rats to scientifically validate its use against diabetes in some parts of India. Ethanolic extract, showing the highest activity in in vitro experiments, was prepared in saline and administered orally to streptozotocin induced albino Wistar diabetic rats for 21 days. Biochemical parameters liver and muscles glycogen and in vivo antioxidant activity in normal, diabetic control, standard (metformin) and treated animals were determined and compared. Attempt was made to isolate, purify and characterize one of the major secondary metabolites in extract by range of chromatographic and spectroscopic techniques. Treatment of streptozotocin induced diabetic rats with ethanolic root extract (500 mg/kg) caused significant (P < 0.01) reduction in blood glucose (312-178 mg/dL), increase in body weight (181-219 g) and serum insulin (1.28-2.26 IU/dL). It also maintained lipid profile and tests of liver and kidney functions within normal range as compared to diabetic control rats and almost at par with standard drug metformin. The oxidative stress induced decline in glutathione and catalase in liver and kidney tissues showed recovery nearly to normal level as a function of treatment. The GC-MS profile of the extract showed relatively high concentration of ß-sitosterol which was characterized by different spectroscopic and chromatographic techniques. The result scientifically and comprehensively validate the reported use of roots of this indigenous plant against diabetes. A strong antioxidant activity of the ethanolic root extract suitably compliments the antidiabetic effect.

A glycoprotein α-amylase inhibitor from Withania somnifera differentially inhibits various α-amylases and affects the growth and development of Tribolium castaneum.

BACKGROUND: Identification and characterisation of plant defensive molecules enrich our resources to design crop protection strategies. In particular, plant-derived proteinaceous inhibitor(s) of insect digestive enzymes appear to be a safe, sustainable and attractive option. RESULTS: A glycoprotein having non-competitive α-amylase inhibitory activity with a molecular weight of 8.3 kDa was isolated and purified from seeds of Withania somnifera α-amylase inhibitor (WSAI). Its mass spectrometry analysis revealed 59% sequence coverage with Wrightide II-type α-amylase inhibitor from Wrightia religiosa. A dose-dependent inhibition of α-amylases from Aspergillus oryzae, Bacillus subtilis, Helicoverpa armigera and Tribolium castaneum was recorded. Interestingly, WSAI did not inhibit human salivary α-amylase significantly. When adults of T. castaneum were fed with WSAI (1.6 mg g-1 ), decrease in consumption, growth and efficiency of conversion of ingested food was evident, along with over fourfold increases in feeding deterrence index. A decline in larval residual α-amylase activity after feeding of WSAI resulted in a reduction in longevity of T. castaneum. CONCLUSION: The study reflects the significance of WSAI in affecting the overall growth and development of T. castaneum. Pre- and post-harvest pest resistive capability makes WSAI a potential candidate for insect pest management. Further, the effectiveness of this inhibitor could be explored either in formulations or through a transgenic approach. © 2016 Society of Chemical Industry.

Isolation and survey of antibiotic sensitivity in nosocomial infections in north Maharashtra region.

BACKGROUND: The present work was aimed to study the prevalence of the drug resistance among Gram positive and Gram negative organisms isolated from clinical samples obtained from various private nursing homes of Jalgaon city and surrounding areas in the state of Maharashtra. MATERIALS AND METHODS: Different pathogenic organisms were isolated from one thousand seven hundred and ninety clinical samples obtained from various nursing homes of Jalgaon region of North Maharashtra, MS, India during the period of August, 2010 to July, 2011. Antibiotic sensitivity testing of isolated pathogenic organisms was done with fifty different antibiotics of 13 different groups by the disk diffusion technique using Mueller Hinton Agar Plates according to CLSI (formerly NCCLS) guidelines. RESULTS: Out of total samples analyzed 1171 samples showed the presence of infection while in 619 samples no growth of organisms was seen on culture medium. Microbial growth revealed presence of 11 different organisms (E. coli, Klebsiella spp., Proteus spp., Pseudomonas spp., Enterobacter spp., Salmonella spp., Acinetobater spp., Streptococccus spp., Staphylococcus aureus (MRSA and MSSA)) including Candida spp. Sensitivity profile of these organisms towards different antibiotics is reported here. CONCLUSION: The study resulted in generation of representative status of the antibiotic sensitivity pattern of commonly found organisms in nosocomial infections in the region. This could be useful for the clinicians to choose correct antibiotic and ensure the judicious use of the same for their patients.

Production of lovastatin by wild strains of Aspergillus terreus.

A wild fungal strain of Aspergillus terreus, labeled as PM3, was isolated by using the Candida albicans bioassay and confirmed by 18S r DNA analyses. Lovastatin was produced by submerged and solid state fermentations. Of the 30 isolated fungal strains, 11 showed lovastatin production with Aspergillus terreus PM3 being the best with a yield of 240 mg/L at the 10th day of submerged fermentation. Carboxymethylcellulose had a stimulatory effect on lovastatin production. It restricted uncontrolled filamentous growth, induced pellet formation and, thereby, improved lovastatin yield. In solid state fermentation (SSF), of the agro wastes from five crops (bran of wheat and rice, husks of red gram and soybean, and green gram straw), wheat bran showed maximum lovastatin production (12.5 mg/g of dry substrate) at pH 7.1 and a temperature of 30 +/- 2 degrees C. Development of a lovastatin production process based on wheat bran as a substrate in SSF is economically attractive as it is a cheap and readily available raw material in agriculture-based countries.

Antidiarrheal activity, chemical and toxicity profile of Berberis aristata.

CONTEXT: This study evaluated the in vitro and in vivo antidiarrheal activity, oral acute toxicological profile, and developed a chemical fingerprint of Berberis aristata Linn. (Berberidaceae). MATERIALS AND METHODS: The ethanol (by maceration) and aqueous (by Soxhlet) extracts of Berberis aristata bark were used for the study. The study involved the antimicrobial (minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) by micro dilution) and antidiarrheal (castor oil induced diarrhea, charcoal motility) tests. The active principle, berberine was characterized by different spectroscopic and chromatographic techniques. RESULTS: The MIC and MBC (of both extracts) against all strains of Shigella were recorded between 125 to 500 µg/mL and 300 to 600 µg/mL, respectively. The MIC and MBC values of berberine are almost comparable to standard ciprofloxacin. UV and IR spectroscopy along with HPTLC and HPLC studies showed presence of berberine in the extracts. The onset of castor oil induced diarrhea was delayed and number of diarrheal episodes was reduced by both the extracts in a dose dependent manner. Similarly, the length of intestine traveled by the feed was also significantly reduced in the charcoal motility test by both the extracts. LD(50) of >5000 mg/kg body weight was observed for both extracts in the acute oral toxicity studies with Swiss albino mice. CONCLUSION: The results validate in vivo and in vitro antidiarrheal activity of Berberis aristata extracts and provide its chemical fingerprint.

Partial characterization and insecticidal properties of Ricinus communis L foliage flavonoids.

Aqueous leaf extract of Ricinus communis L (Euphorbiaceae), a cultivated plant in tropical countries, showed excellent insecticidal activity against Callosobruchus chinensis L (Coleoptera: Bruchidae). We have isolated and tested flavonoids as insecticidal and antimicrobial agents. The isolated flavonoids showed potential insecticidal, ovicidal and oviposition deterrent activities against C chinensis L. However, antimicrobial activity against the common microbial infestants of stored pulses, of which C chinensis is a major pest, was found to be insignificant. Two bands having Rf 0.63 and 0.69 were seen on HPTLC plates using mobile phase benzene + ethyl acetate + methanol + formic acid (12 + 4.5 + 2 + 1.5 by volume) as eluant. The Rf values and scanning of the spectrum in the ultraviolet region, showed identity with the flavonoids quercetin and kaempferol. This was further confirmed using HPLC and IR and UV spectrometry. HPLC and HPTLC chromatograms also suggested quercetin to be the major flavonoid present in the hydrolyzed aqueous leaf extract of R communis.

Antimicrobial and pesticidal activity of partially purified flavonoids of Annona squamosa.

Foliar extracts of Annona squamosa (Family: Annonaceae) were screened for antimicrobial and insecticidal activity against the common microbial infestants of pulses and the stored grain pest pulse beetle, Callosobruchus chinensis (Coleoptera: Bruchidae). Flavonoids isolated from aqueous extracts of A squamosa showed antimicrobial activity against all the common microbial contaminants of pulses and 80% insecticidal activity against C chinensis at a concentration of 0.07 mg ml-1. Various physico-chemical tests, chromatographic and spectroscopic studies with partially purified aqueous extract indicated the presence of flavonol type flavonoids. This may provide a useful beginning for the development of botanical pesticides for post-harvest safeguard of pulses.