Extraction of bulk DNA from Thar Desert soils for optimization of PCR-DGGE based microbial community analysis

A reliable method for characterizing microbial communities on the basis of their differences in the 16S ribosomal RNA (rRNA) gene sequences in the hot arid zone sandy soils has been optimized. A desert plant ( Calligonum polygonoides ) was chosen to provide the rhizospheric soil samples, collected from three different agro-ecological locations. Total community DNA was efficiently extracted at small-scale level using direct lysis with hot sodium dodecyl sulphate (SDS), glass bead beating and finally subjecting the sandy soil to liquid nitrogen freeze-thaw cycles. To amplify V3 region of bacterial 16S rRNA gene, universal conserved primers were used. Second round polymerase chain reaction (PCR) was attempted to increase product concentration and to minimize the effect of inhibitory substances. To enhance the detection sensitivity of the denaturing gradient gel electrophoresis (DGGE), the effect of change in template DNA concentration was studied. The separation of bands were greatly enhanced in the fingerprints obtained after the second round of PCR representing low abundant species which were not differentiated at single optimized concentration of DNA

Sinteza, spazmolitsko i antidijaretsko djelovanje nekih 1-supstituiranih derivata imidazola

A series of 1-substituted imidazoles 1a-d and 2a-d were synthesized and screened for antispasmodic and antidiarrheal activities. Antispasmodic activity was tested at various concentrations on isolated tissue preparations; concentration-response curves were plotted and compared with atropine. All compounds were found to inhibit contraction of the guinea pig ileum. Castor oil-induced diarrhea model in rats was used for evaluation of antidiarrheal activity. Parameters such as intestinal transit and volume of intestinal fluid were measured for antidiarrheal activity at 40 mg kg-1 dose and compared with the standard drug loperamide at a 6 mg kg1 dose. Defecation frequency in the test group was found to be significantly lower (p < 0.01) compared to the control group and comparable with that of the standard. The present study reveals that the compounds exert antidiarrheal activity through possible inhibition of intestinal movement and reduction of capillary permeability in the abdominal cavity.U radu je opisana sinteza serije novih 1-supstituiranih imidazola 1a-d i 2a-d te evaluacija njihovog spazmolitskog i antidijaretskog djelovanja. Spazmolitsko djelovanje različith koncentracija ispitivanih spojeva (10, 50 i 100 µg mL1) ispitano je na izoliranim tkivnim preparatima; krivulje ovisnosti djelovanja o koncentraciji uspoređene su s djelovanjem atropina. Svi ispitivani spojevi inhibirali su kontrakcije ileuma zamorca. Dijarea inducirana ricinusovim uljem u štakora korištena je za vrednovanje antidijaretskog djelovanja. Intestinalni prijenos i volumen intestinalne tekućine mjereni su pri dozi od 40 mg kg1 i uspoređeni s učinkom standarda loperamida (doza 6 mg kg1). Učestalost defekacije u testiranoj skupini bila je značajno niža (p < 0,01) u usporedbi s kontrolnom skupinom i standardom. Provedena ispitivanja pokazuju da antidijaretsko djelovanje ispitivanih spojeva može biti posljedica inhibicije intestinalnih pokreta i redukcije kapilarne permeabilnosti u abdominalnoj šupljini

GCMS analysis and antimicrobial action of latex of <i>Euphorbia caducifolia</i>

Euphorbia caducifolia is conspicuously and regularly represented in the flora of the Rajasthan state. Latex of E. caducifolia is used to cure skin infections, cutaneous eruption, leucoderma and applied to cuts and wounds for speedy healing. The GCMS analysis of fraction isolated from latex showed presence of methyl palmitate, 5,9-heptadecadienoate, methyl 11 octadecenoate, methyl octadecenoate and 3,7,11,15-tetramethyl- 2-hexadecene-l-ol. Isolated fraction of E. caducifolia (IFEC) and latex of E. caducifolia (ECL) were tested against S. aureus, M. luteus, B. subtilis, E. coli, S. typhi, A. niger and C. albicans. IFEC was found to be more effective against fungal species, and MIC was found to be 150 and micro;g/ml against A. niger. [J Intercult Ethnopharmacol 2012; 1(2.000): 119-123

Study on the effect of different parts of Nyctanthes arbortristis linn. on the hematological parameters of mice

Hematological effect of the ethanolic extracts of the flowers, barks, seeds and leaves of Nyctanthes arbortristis Linn were estimated at the dose levels of 200, 400 and 600 mg/kg body weight (I.P.), which were selected from the study of their LD50 value. From the overall study it was observed that the ethanolic extracts have antianemic, hematostatic effect and are hematologically nontoxic. Whereas the ethanolic extracts of the seeds and leaves at the dose of 600mg/kg may possess immunostimulatory effect

Antidiabetic Activity of Pterospermum acerifolium Flowers and Glucose Uptake Potential of Bioactive Fraction in L6 Muscle Cell Lines with Its HPLC Fingerprint

The present study was designed to estimate the detailed antidiabetic activity of Pterospermum acerifolium (L.) Willd flowers. In vitro alpha amylase inhibition study was carried out on 50% ethanol extract of flowers (PAFEE) and its various fractions. The active ethyl acetate fraction (PAFEF) was subfractionated into three subfractions (PAFE1, PAFE2, and PAFE3) and subjected to acute toxicity studies followed by antidiabetic screening in vivo by streptozotocin-nicotinamide induced type II diabetes. Diabetic animals treated with PAFE2 (30 mg/kg) reduced the levels of fasting blood glucose, significantly (P<0.001) compared to that of diabetic control animals. Histological studies on drug treated groups did not show remarkable positive changes in β-cells. PAFE2 showed 32.6±1.93% glucose uptake over control and, in the presence of PI3K inhibitor wortmannin, declined to 13.7±2.51%. HPLC analysis of PAFE2 reveals the presence of quercetin and apigenin as major constituents and both are inhibiting the glycogen phosphorylase enzyme in molecular modelling studies. The study evidenced strongly that the probable glucose lowering mechanism of action of active subfraction PAFE2 is by increasing the glucose uptake in peripheral tissues and by inhibition of gluconeogenesis

Extraction of bulk DNA from Thar Desert soils for optimization of PCR-DGGE based microbial community analysis

A reliable method for characterizing microbial communities on the basis of their differences in the 16S ribosomal RNA (rRNA) gene sequences in the hot arid zone sandy soils has been optimized. A desert plant ( Calligonum polygonoides ) was chosen to provide the rhizospheric soil samples, collected from three different agro-ecological locations. Total community DNA was efficiently extracted at small-scale level using direct lysis with hot sodium dodecyl sulphate (SDS), glass bead beating and finally subjecting the sandy soil to liquid nitrogen freeze-thaw cycles. To amplify V3 region of bacterial 16S rRNA gene, universal conserved primers were used. Second round polymerase chain reaction (PCR) was attempted to increase product concentration and to minimize the effect of inhibitory substances. To enhance the detection sensitivity of the denaturing gradient gel electrophoresis (DGGE), the effect of change in template DNA concentration was studied. The separation of bands were greatly enhanced in the fingerprints obtained after the second round of PCR representing low abundant species which were not differentiated at single optimized concentration of DNA

Antioxidant and DNA damage protective properties of anthocyanin-rich extracts from <i>Hibiscus</i> and <i>Ocimum</i>: a comparative study

<div><p>Anthocyanin extracts (AEs) from <i>Ocimum tenuiflorum</i> (leaf), <i>Hibiscus rosa-sinensis</i> (petal) and <i>Hibiscus sabdariffa</i> (calyx) were investigated and compared for <i>in vitro</i> antioxidant activity and DNA damage protective property. Total phenolic content (TPC) and total anthocyanin content (TAC) of the AEs were determined and the major anthocyanins were characterised. <i>In vitro</i> antioxidant activities were assessed by ferric-reducing antioxidant power (FRAP) assay, 2,2-diphenyl-1-picryl hydrazyl (DPPH) radical-scavenging activity, 2-deoxy-d-ribose degradation assay and lipid peroxidation assay. The protective property of the AEs was also examined against oxidative DNA damage by H₂O₂ and UV using pUC19 plasmid. All the AEs particularly those from <i>O. tenuiflorum</i> demonstrated efficient antioxidant activity and protected DNA from damage. Strong correlation between antioxidant capacity and TPC and TAC was observed. Significant correlation between antioxidant capacity and TPC and TAC ascertained that phenolics and anthocyanins were the major contributors of antioxidant activity.</p></div