Abstract  The antioxidant activities of Methanol/Chloroform (MC) extracts and fractions of five brown algae (Sargassum fusiforme, Sargassum kjellmanianum, Sargassum. pallidum, Sargassum thunbergii and Sargassum horneri) from China were evaluated through DPPH/hydroxyl radical-scavenging activity and reducing power. Total phenolic content was investigated using Folin-Ciocalteau reagent. The MC extract of S. kjellmanianum showed higher antioxidant activity than other seaweeds. However, the MC extracts of all the seaweeds showed lower DPPH radical-scavenging activity than Gallic Acid (GA) and ascorbic acid (AscA) at 50 mu g/ml. The MC extracts were further purified by liquid-liquid partition to afford four fractions: Petroleum Ether (PE), Ethyl Acetate (EA), Butanol (BU) and Aqueous (AQ), among which the EA was found to be the most effective fraction and showed higher antioxidant activity than AscA, except that the EA fraction of S. thunbergii presented lower reducing power than GA. Antioxidant activities of MC extracts showed an increase with increasing concentration (between 15 and 45 mu g/ml) indicating the dose dependency of these properties. The phenolic compounds with highest activities were concentrated in the EA fraction and were of medium polarity. The finding showed that the antioxidant-rich extracts or fractions had a significant potential to be use as a natural antioxidant agent.

Abstract  Epidemiologic studies of disinfection by-products have traditionally focused on total trihalomethane (TTHM) concentration as a surrogate for maternal exposure during pregnancy. We used birth certificate data on 196,000 infants to examine the effect of third-trimester exposures on various indices of fetal development. We examined the effect of town-average concentrations of TTHM and additional exposure metrics in relation to mean birth weight, mean gestational age, small for gestational age (SGA) infancy, and preterm delivery. Trihalomethane data (TTHM, chloroform, and bromodichloromethane) from 1995-1998 were available for 109 towns in Massachusetts. Data from 1997-1998 on haloacetic acid (total haloacetic acids, dichloroacetic acid, and trichloroacetic acid), 3-chloro-4-(dichloromethyl)-5- hydroxy-2(5H)-furanone (MX), and mutagenicity were available for a limited number of towns. We observed reductions in mean birth weight (12-18 g) for maternal trihalomethane exposures > the 90th percentile compared with those < the 50th percentile. Birth weight reductions were detected for chloroform exposures > 20 microg/L and TTHM exposures > 40 microg/L. Elevated trihalomethanes were associated with increases in gestational duration and a reduced risk of preterm delivery. We found evidence of an exposure-response effect of trihalomethanes on risk of SGA, with odds ratios (ORs) ranging from 1.09 to 1.23 for bromodichloromethane exposures > 5 microg/L. Elevated mutagenic activity was associated with SGA [OR = 1.25; 95% confidence interval (CI), 1.04 to 1.51] and mean birth weight (-27 g; 95% CI, -54 to -1). Although smaller in magnitude, our findings are consistent with previous studies reporting associations between trihalomethanes and SGA. These data also suggest a relationship between fetal development indices and mutagenic activity independent of exposure to trihalomethanes, haloacetic acids, and MX.

Abstract  Chloroform is being administered to male Osborne-Mendel rats and to female B6C3F1 mice at concentrations of 0 (negative control), 200, 400, 900 or 1800 ppm in the drinking water. Matched control groups of both species receive a volume of water identical to that consumed by the corresponding 1800 ppm groups. At this writing, the animals have completed 23 months on test. Negative control and CHCl3-treated rat groups have shown typical growth curves, with dose-related relative decrements in body weight evident throughout the study. Following decreases in CHCl3 groups during the first 8 weeks, rat water consumption values have continued to increase slowly, but persistent relative dose-related decrements are evident. No initial treatment-related decrements are evident. No initial treatment-related mortality was seen in the rats. Survival is 21, 41, 45, 76, 70 and 64% for the negative control, 200, 400, 900, 1800 ppm and matched control groups, respectively. Survival values for mice at three weeks were 99, 94, 74 and 76% for the 200, 400, 900 and 1800 ppm groups, respectively. Mortality was apparently related to markedly decreased fluid consumption among some of the treated mice. Subsequent mortality has been less than 15% for all mouse groups. Except for acclimation effects during the first 2-3 weeks, body weights for the treated mouse groups have been generally within 10% of negative control values. Tissue changes in decedents have been similar in treated and control groups, both in rats and mice. Terminal sacrifice and histologic evaluations will be initiated after completion of 24 months on test.

Abstract  Given that there are many autopsy cases in which erectile dysfunction (ED) treatment drugs can be detected from elderly men who are diagnosed to have died of cardiovascular disorders, the degree of cardiovascular risk posed by ED treatment drugs is still controversial. Moreover, counterfeit ED drugs or illegal dietary supplements containing ED drugs are also threats to public health. In this study, we established a detailed procedure for simultaneous analysis of typical ED drugs (sildenafil, vardenafil, tadalafil) and their metabolites in human blood and urine by isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS-MS). Each sample of whole blood and urine containing the three ED treatment drugs, their metabolites, and deuterated internal standards (ISs) was diluted with alkalinized water, loaded onto an Oasis HLB cartridge, washed with dilute ammonium hydroxide solution, and eluted with chloroform. The eluate was acidified with methanol and concentrated HCl and evaporated to dryness. The resulting residue was reconstituted with methanol and mobile phase solution, and 5 mu l of the solution was injected into an LC-MS-MS instrument. The determinations were made by multiple reaction monitoring using each product ion. The recovery rates of the drugs, metabolites, and ISs from whole blood and urine ranged from 80.7 to 127%. Good linearity was obtained for all drugs and their metabolites in the range of 1-100 ng/ml in whole blood and urine with correlation coefficients greater than 0.99. The detection limits (signal-to-noise ratio = 3) for all compounds were not higher than 0.05 ng/ml. Intraday and interday accuracy and precision data were also satisfactory for all compounds in whole blood and urine. Actual measurements of sildenafil and N-desmethyl sildenafil were also demonstrated by analysis of whole blood and urine specimens from two male volunteers after ingestion of a 25-mg tablet of sildenafil.

Abstract  The antimicrobial activity and phytochemicals of the leaves and bark of Vitex negundo L. was evaluated against three Gram-positive bacteria viz. Staphylococcus epidermidis, Bacillus subtilis, Staphylococcus aureus and five Gram-negative bacteria viz. Escherchia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Vibrio cholerae and Vibrio alginolyteus. Both polar and nonpolar extracts viz. petroleum ether, chloroform, ethanol, methanol and aqueous extracts were prepared and studied for antibacterial activity using disc diffusion, agar cup and broth dilution methods. Results showed promising antibacterial activity of all the extracts of both leaf and bark against E. coli, followed by S. aureus. Ethanol and methanol extracts of the leaf showed inhibition activity against both Gram-positive and Gram-negative bacteria where as petroleum ether and chloroform extracts of bark had better antibacterial activity against Gram-positive bacteria. MIC showed that 5% of the extracts were active in a concentration of 0.312 mg/ml; 27.5% in a concentration of 0.625 mg/ml and 88.75% in a concentration of 1.25 mg/ml; were active against different human pathogenic bacteria. At concentration about 2.5 mg/ml, 100% of inhibition was recorded against both Gram-positive and Gram-negative bacteria. The result obtained with ethanol and methanol extracts of leaves; petroleum ether and chloroform extract of bark exhibited significant antibacterial activity, a property that supports traditional use of the plant in the treatment of some diseases as broad spectrum antibacterial agents.

Abstract  BACKGROUND: Scaevola spinescens is an endemic Australian native plant with a history of use as a medicinal agent by indigenous Australians. Yet the medicinal bioactivities of this plant are poorly studied.

MATERIALS AND METHODS: S. spinescens solvent extracts were tested for antimicrobial activity, antiviral activity and toxicity in vitro.

RESULTS: All extracts displayed antibacterial activity in the disc diffusion assay. The methanol extract proved to have the broadest specificity, inhibiting the growth of 7 of the 14 bacteria tested (50%). The water, ethyl acetate, chloroform, and hexane extracts inhibited the growth of 6 (42.9%), 5 (35.7%), 5 (35.7%), and 4 (28.6%) of the 14 bacteria tested, respectively. S. spinescens methanolic extracts were equally effective against Gram-positive (50%) and Gram-negative bacteria (50%). All other extracts were more effective at inhibiting the growth of Gram-negative bacteria. All extracts also displayed antiviral activity in the MS2 plaque reduction assay with the methanol, water, ethyl acetate, chloroform, and hexane extracts inhibiting 95.2 ± 1.8%, 72.3 ± 6.3%, 82.6 ± 4.5%, 100 ± 0% and 47.7 ± 12.9% of plaque formation, respectively. All S. spinescens extracts were nontoxic in the Artemia fransiscana bioassay with no significant increase in mortality induced by any extract at 24 and 48 h. The only increase in mortality was seen for the water extract at 72 h, although even this extract displayed low toxicity, inducing only 41.7 ± 23.3% mortality.

CONCLUSIONS: The lack of toxicity of the S. spinescens extracts and their inhibitory bioactivity against bacteria and viruses validate the Australian Aboriginal usage of S. spinescens and indicates its medicinal potential.

Abstract  Sea buckthorn berries are therapeutically used as folk medicine for a variety of diseases, however, the scientific evidence is hardly available to support their role. This study explored their chemical constituents and their role as antioxidant and antibacterial agents. Three common solvents such as petroleum ether (40° - 60°C), chloroform and methanol were successively used for the extraction of active principles from sea buckthorn berries. Five major fractions (F1-F5) were isolated from the active methanol extract by column and thin layer chromatography. An attempt was made to identify the chemical nature of pooled fractions by available spectral means. Antioxidant potential of methanol extract and its fractions was measured by DPPH, formation of phosphomolybdenum complex and TBA methods. The hole-plate diffussion method was used to find out the antibacterial activity. A very brief structure-activity relationship of the potent antioxidant and antimicrobial compounds is discussed. Methanolic extract and its fractions contain numerous phenolic compounds such as flavonoids, which may be responsible for antioxidant and antibacterial effects.

Abstract  During screening for antibiotic producing microorganisms from environmental soil samples, the supernatant of a bacterial isolate was found to have antibacterial and antifungal activity on the standard indicator species. The standard cylinder-plate method was used to determine the inhibitory effect of the crude supernatant of each isolate on 6 bacterial and 3 fungal standard strains by measuring the diameter of inhibition zone. The highest inhibition zone on Aspergillus niger belonged to culture broth of isolate FAS1 by 25 mm, and this isolate was the most efficient microorganism to inhibit standard bacterial and fungal species. Based on morphological and biochemical properties as well as 16S rDNA gene analysis, the selected isolate (isolate FAS(1)) belonged to Bacillus genus. Investigation on the ability of different culture media for antibiotic production led to select Luria-Bertani media for further studies. Treatment of the culture broth of the isolate FAS(1) using typical protease didn't decrease the antimicrobial activity of the supernatant. After extracting of culture broth of the selected isolate by ethyl acetate as an organic solvent, the inhibitory effect was mainly increased. More investigation was done by bioautography method where the ethyl acetate fraction of the broth culture was separated on TLC by chloroform:methanol, 60:40 as mobile phase and R(f) were calculated for inhibition spots.

Abstract  Context: Medicinal plants are a largely unexplored source of drug repository. Urtica dioica L. (Urticaceae) is used in traditional medicine to treat diverse conditions. Objective: The present study describes the antidiabetic, antiinflammatory, antibacterial activity, and toxicological studies of Urtica dioica. Materials and methods: U. dioica leaves were subjected to solvent extraction with hexane, chloroform, ethyl acetate, methanol, and aqueous, respectively, and screened for antidiabetic (300 mg/kg bw by glucose tolerance test; GTT), antiinflammatory (200 mg/kg bw by rat paw edema assay) and antibacterial activities [by disc-diffusion and minimum inhibitory concentration (MIC) assays]. Toxicological studies were carried on Artemia salina and Wistar rats; phytochemical analyses were carried out, using chromatographic and spectroscopic techniques. Results: The aqueous extract of U. dioica (AEUD) significantly (p < 0.001; 67.92%) reduced the blood glucose level during GTT in Wistar rats with an effective dose of 300 mg/kg bw in dose-dependent studies. High-performance liquid chromatography with photodiode-array detection (HPLC-DAD) analysis showed the presence of hydroxycinnamic acid derivatives and flavonoids in AEUD. Hexane Fraction-2 (HF2) exhibited both antiinflammatory activity (48.83% after 3 h), comparable to that of indomethacin (53.48%), and potent antibacterial activity with MIC values ranging from 31.25-250 µg/mL against all the tested strains. Gas chromatography-mass spectrometry (GC-MS) analysis showed fatty acid esters and terpenes as the major constituents of HF2. Toxicity tests showed higher safety margin of all the solvent extracts with LC(50) > 1000 μg/mL each on A. salina. Discussion and conclusion: Our results showed that the U. dioica leaves are an interesting source of bioactive compounds, justifying their use in folk medicine, to treat various diseases.

Abstract  OBJECTIVE: To determine adulticidal activitiy of hexane, ethyl acetate, benzene, chloroform and methanol leaf extracts of Cardiospermum halicacabum against Culex quinquefasciatus (Cx. quinquefasciatus), Aedes aegypti (Ae. aegypti) and Anopheles stephensi (An. stephensi).

MATERIALS AND METHODS: The bioassay was conducted in an experimental kit consisting of two cylindrical plastic tubes both measuring 125 x 44 mm following the WHO method; Mortality of the mosquitoes was recorded after 24 h.

RESULTS: The adulticidal activity of plant extracts showed moderate toxic effect on the adult mosquitoes after 24 h of exposure period. However, compared to other solvents highest mortality was found in methanol extract of C. halicacabum against all the three mosquitoes. Among them An. stephensi produce the highest LC50 and LC90 (186.00 and 346.06 ppm) values.

CONCLUSIONS: From the results it can be concluded the crude extract of C. halicacabum was an excellent potential for controlling Cx. quinquefasciatus, Ae. aegypti and An. stephensi mosquitoes.

Abstract  ETHNOPHARMACOLOGICAL RELEVANCE: The present investigation was aimed to justify the pharmacological basis in traditional use of Clerodendrum colebrookianum as antihypertensive agent in north-east India.

MATERIALS AND METHODS: The aqueous extract (AECc), its aqueous, n-butanol (nBFCc), Ethyl-acetate (EtFCc) and Chloroform fractions of C. colebrookianum leaves were evaluated for antihypertensive potential by using fructose-induced hypertension model in rats and in isolated frog heart. The ex-vivo muscarinic action in isolated rat ileum, in-vitro assay for Rho-kinase (ROCK -II), phosphodiesterase-5 (PDE-5) and angiotension converting enzyme (ACE) were also carried out to establish the mechanism of action of samples. The total phenolic and flavonoied contents in test samples were estimated to establish phyto-pharmacological relationship.

RESULTS: The 100μg/mL test samples were showed calcium antagonism in rat ileum and at 50μg/mL and 75μg/mL doses exhibited ROCK-II and PDE-5 inhibition respectively where, EtFCc was caused maximum 68.62% (ROCK-II) and 52.28% (PDE-5) inhibition, but none of the sample was exhibit effect in ACE at 100μg/mL. The test samples also showed negative inotropic and chronotropic effect on isolated frog heart and significant (P<0.001) reduction in systolic blood pressure and heart rate in hypertensive rats compared to control. The total phenolic content maximum 80μg gallic acid equivalents in nBFCc and flavonoids content maximum 69.57μg Quercetin equivalent in AECc were estimated.

CONCLUSIONS: These observations established the traditional claim and thus C. colebrookianum could be a potent antihypertensive agent for use in future. The antihypertensive effect mediated by cholinergic action and following ROCK - II, PDE-5 inhibition of C. colebrookianum.

Abstract  Lipid extraction from biological samples is a critical and often tedious preanalytical step in lipid research. Primarily on the basis of automation criteria, we have developed the BUME method, a novel chloroform-free total lipid extraction method for blood plasma compatible with standard 96-well robots. In only 60 min, 96 samples can be automatically extracted with lipid profiles of commonly analyzed lipid classes almost identically and with absolute recoveries similar or better to what is obtained using the chloroform-based reference method. Lipid recoveries were linear from 10-100 µl plasma for all investigated lipids using the developed extraction protocol. The BUME protocol includes an initial one-phase extraction of plasma into 300 µl butanol:methanol (BUME) mixture (3:1) followed by two-phase extraction into 300 µl heptane:ethyl acetate (3:1) using 300 µl 1% acetic acid as buffer. The lipids investigated included the most abundant plasma lipid classes (e.g., cholesterol ester, free cholesterol, triacylglycerol, phosphatidylcholine, and sphingomyelin) as well as less abundant but biologically important lipid classes, including ceramide, diacylglycerol, and lyso-phospholipids. This novel method has been successfully implemented in our laboratory and is now used daily. We conclude that the fully automated, high-throughput BUME method can replace chloroform-based methods, saving both human and environmental resources.

Abstract  Biological screening of Scrophularia nodosa crude extract and its fractions (hexane, chloroform, ethyl-acetate, n-butanol and aqueous) was carried out on phytotoxicity, cytotoxicity, antibacterial, antifungal and analgesic activities. Crude extract and its fractions produced 50-100% phytotoxicity at 1000μ/mL concentration whereas 25-77% phytotoxicity was observed at 10μ/mL concentration. The fractions exhibited significant antibacterial and antifungal effects. The non-toxic results of this plant were recorded in Brine Shrimps Bioassay method at all concentrations. Similarly no significant insecticidal activity was observed in crude extracts and fractions. Analgesic activity results of S. nodosa in mice were found highly significant in crude extract as compared to fractions. In writhing test crude extract at 500 mg/kg showed 65.6% highest inhibitory response in mice.

Abstract  CONTEXT: Carpolobia lutea G. Don (Polygalaceae) leaf is reputable as an antidiarrheal agent among the Efik and Ibibio tribe of Akwa Ibom State, Nigeria. The crude extract is reported to show antidiarrheal and antiulcer effects in rodents.

OBJECTIVE: The isolation and characterization of drug molecules from the leaf fraction with antidiarrheal bioactivity and determination of mechanism of action are reported.

MATERIAL AND METHODS: Gradient extraction by maceration yielding n-hexane, chloroform, ethyl acetate, and ethanol fractions (770 mg/kg) were used to establish the fractions suitable for drug discovery. The antidiarrheal effect of the leaf fractions of Carpolobia lutea was evaluated using castor oil-induced diarrhea, castor oil-induced intestinal transit, and enteropooling.

RESULTS: Results indicate that all fractions produced a significant (p < 0.01-0.001) decrease in castor oil-induced diarrhea in rats. This effect was not antagonized by isosorbide dinitrate (150 mg/kg, p.o), diphenoxylate (5 × 10⁻³ mg/kg p.o) and yohimbine (1 mg/kg, s.c.) except for the chloroform fraction. The ethyl acetate fraction produced 100% inhibition of intestinal transit, an effect greater than pure drug. Phytochemical analysis of the ethyl acetate fraction yielded polyphenolic compounds.

CONCLUSION: The leaf fractions contain two types of antidiarrheal agents, one mediating its effect through α₁-presynaptic adrenoceptor while the other does not. Polyphenols isolated may in part lend credence for observed antidiarrheal activity.

Abstract  Psoralea corylifolia (Leguminosae) is traditionally used for the treatment of skin conditions, fever and as anthelmintic. The aim of present study was to standardize the crude extract of P. corylifolia and determine its pharmacological activities. The crude extract of P. corylifolia was standardized by high performance liquid chromatography (HPLC), Fourier transform infra red (FT-IR), Fourier transform near infra red (FT-NIR) and ultra violet (UV) spectrophotometric methods. Pharmacological study of the crude extract of P. corylifolia and its fractions were carried out on smooth muscle of rabbit intestine. The results revealed quick decrease in the normal intestinal movement followed by a gradual dose dependant increase in the rhythmic activity of intestine. The antispasmodic response of the crude extract was found most significant at 20 mg/ml (78.13%). The chloroform fraction of the crude extract exhibited maximum antispasmodic response at 10 mg/ml (45.16%). n-butanol and aqueous fractions produced 39.29 and 13.79% antispasmodic effect, respectively, whereas, ethyl acetate fraction produced spasmogenic effect (21.9%). The crude extract of P. corylifolia exhibited positive antifungal activity against Candida albicans and positive antibacterial activity against Staphylococcus aureus. Significant analgesic effect was also observed with 300 and 500 mg/kg dose at P < 0.05. The analgesic effect of the plant extract (500 mg/kg) was found to be higher than that of diclofenac sodium (50 mg/kg; positive control). Brine shrimp lethality assay was performed and the plant extract did not exhibit any toxicity.

Abstract  Euphorbia wallichii a perennial herb growing mainly in Himalayas has been widely used in folk medicines for its medicinal properties. In the present study, the crude methanolic root extract (CME) and its fractions; n-Hexane Fraction (NHF), n-Butanol Fraction (NBF), Chloroform Fraction (CHF), Ethyl acetate Fraction (EAF) and Aqueous Fraction (AQF) of this plant specie were investigated for antioxidant and cytotoxic activities and phytochemical analysis. Antioxidant activity was determined by using 2,2-diphenyl-1-picryl-hydrazyl free radical (DPPH) and DNA protection assay performed on pBR322 plasmid DNA. In both these assays, promising results were obtained for CME as well as other fractions. The IC50 values for DPPH assay were in a range of 7.89 to 63.35 mu g/ml in which EAF showed the best antioxidant potential and almost all the tested samples showed certain level of DNA protection. The cytotoxic activity was assessed by using Sulforhodamine B (SRB) assay on human cell lines; H157 (Lung Carcinoma) and HT144 (Malignant Melanoma). The IC50 values of the tested samples ranged from 0.18 to 1.4 mg/mL against H157 cell line whereas against HT144 cell line the IC50 values ranged from 0.46 to 17.88 mg/mL with NBF fraction showing maximum potential for both. Furthermore, the phytochemical analysis of CME and its fractions showed the presences of flavonoids, saponins, tannins, terpenoides and cardiac glycosides with varying concentrations.

Abstract  Petroleum ether, ethyl acetate, chloroform, butanol and methanol extracts of the leaves of Marrubium alysson L. were screened for in vitro antiviral activity against human cytomegalovirus (HCMV) strain AD-169 (ATCC Ref. VR 538) and coxsackie B virus type 3 (CoxB-3) using a cytopathic effect (CPE) reduction assay. Total phenolic content of the extracts were determined by the Folin-Ciocalteu colorimetric method. The most potent inhibition was observed with methanol extract, which inhibited HCMV and Cox B3 viruses replication at 100 mu g/ml without showing cytotoxic effects. Good activities were also found with the ethyl acetate, butanol and chloroform extracts which exhibited antiviral effect against both HCMV and Cox B3 at 100 to 250 mu g/ml.

Abstract  The seed products of Carica papaya have been proven as potential male contraceptives in laboratory animals. In this study, chromosomal aberrations were investigated in spermatogonia of albino rats and rabbits, following oral administration of Methanol Sub-Fraction (MSF) of Carica papaya seeds. The experimental animals were divided into three groups, each group had 5 male Wistar rats and rabbits. In the first group, double distilled water served as negative control. The second group received monomeric acrylamide at 72.5 mg kg(-1) body weight; two doses with a gap of 3 h served as positive control and in the third group, the MSF was orally administered at 500 mg kg(-1) body weight (10 x contraceptive dose; CD); two doses with a gap of 3 h. The chromosomal fragments, dicentrics, rings, exchanges, damaged chromosomes and total chromosomal aberrations in MSF treated rats and rabbits were not significantly different when compared with negative control animals, however, were found to be reduced significantly (p<0.001) when compared with positive control group. The results suggested that the MSF did not induce chromosomal aberrations.

Abstract  The present investigation was designed to evaluate the antibacterial, antifungal properties and physiochemical screening of the various leaves and stem extracts of Andrographis lineata. Acetone, ethanol, methanol, petroleum ether and chloroform extracts of shade dried plant leaf and stem of A. lineata were tested for antibacterial, antifungal and phytochemical analysis. The antibacterial properties of various extracts of leaves and stem of A. lineata were assayed using the standard disc diffusion method, against five strains of bacterial species, namely, Proteus vulgaris, Escherichia coli, Klebsiela pneumoniae, Staphylococcus aureus and Pseudomonas aeruginosa. Among different solvent extracts studied, the methanol leaf extract showed highest antibacterial activity against staphylococcus aureus (19.41 mm); followed by petroleum ether extract (18.30 mm). The highest inhibition zone observed for acetone extract of A. lineata leaves against P. vulgaris was 17.40 mm followed by ethanol extract (16.10 mm). The highest activity of chloroform extract against K. pneumoniae was 14.07 mm. Phytochemical investigation confirmed the presence of flavonoids, saponins, gums and mucilages, triterpenoids, steroids, glycosides, phenolic compounds and tannins. The acetone leaf extracts of A. lineata showed highest antifungal activity against Penicillium pinophilum (13.40 mm). The methanol leaf extracts showed maximum activity against Aspergillus niger (12.15 mm). The petroleum ether leaf extracts showed highest activity against P. pinophilum (12.11 mm). The ethanol leaf extracts showed significant activity against Aspergillus flavus (11.24 mm). The chloroform leaf extracts showed higher activity against A. niger (9.75 mm). The leaf extract showed more inhibitory effect than the stem extracts. The present research justifies the claimed uses of this herb in the traditional system of medicine to treat different diseases.

Abstract  The present study shows the phytochemical analysis and antibacterial activity of leaf and callus of Centella asiatica. Leaf explants of C. asiatica were cultured on MS medium supplemented with different concentration of plant growth regulators for callus initiation. The maximum percentage of callusing was achieved in medium supplemented with 6-benzylaminopurine 4.0 mg/Land 2,4-dichlorophenoxyacetic acid 2.0 mg/L. In the preliminary phytochemical screening, alkaloids, glycosides, terpenoids, steroids, flavonoids, tannins, saponins and reducing sugars were present in most of the tested extracts of leaf and in vitro grown callus of C. asiatica. Methanol, acetone, chloroform and water extracts of leaf and callus were evaluated for in vitro antibacterial activity against Bacillus cereus, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa by agar plate well diffusion method. All the extracts from leaf and callus of C. asiatica showed significant antibacterial activity against the tested organisms. However, methanol extracts of leaf and callus showed maximum inhibitory effect.

Abstract  Physalis minima is an important medicinal plant of Indian System of Medicine. This plant is reported for its diuretic, laxative and antiinflammatory activities. However, the plant is not well scrutinized for its antimicrobial potential. The major chemical constituents reported from the plant are phenolics and alkaloids, which suggest that the plant may turn out to be a potent antiinfective agent. The aim of the study was to find out the antibacterial potential of mature berries of P. minima using streak plate, well diffusion, determination of minimum inhibitory concentration and bioautographic methods against a battery of Gram positive and Gram negative bacterial strains. Results of the study showed that methanol and chloroform extracts of P. minima exhibited potent inhibitory activity against all the bacterial strains tested. Minimum inhibitory concentration found out was 100 μg in both the extracts. Bioautography assay showed polar compounds present in the crude extract are responsible for the antimicrobial action.

Abstract  Methanol, aqueous and chloroform extracts of Cleome viscosa and Cleome burmanni were tested for anthelmintic potential against the Indian earthworm Pheritima posthuma. Different concentrations of the extracts ranging from 50-2000 μg/ml were tested and results expressed as time required for paralysis and death of the worms. Piperazine citrate was used as a reference standard and DMSO (1%) as the negative control. The methanol extracts of Cleome viscosa and Cleome burmanni exhibited significant anthelmintic activity. Methanol extract of Cleome viscosa at a concentration of 2000 μg/ml was detected to be the most effective treatment dose. Thin layer chromatography of methanol extracts of both plants revealed the presence of terpenoids.

Abstract  CONTEXT: Eucalyptus camaldulensis Dehnh. (Myrtaceae) and Eucalyptus torelliana F. Muell are used in Nigerian traditional medicine for the treatment of cough associated with tuberculosis (TB) and other respiratory infections.

OBJECTIVE: Hexane, chloroform, methanol extracts, and isolated compounds of E. camaldulensis and E. torelliana were screened for activity against Mycobacterium tuberculosis H37Rv (MtbH37Rv) to authenticate the traditional use of these plants.

MATERIALS AND METHODS: The microplate alamar blue assay (MABA) method was used to investigate the anti-M. tuberculosis activities. Bioassay-guided fractionation of the hexane extract of E. torelliana leaf was performed, and isolated compounds were characterized by MS, 1D- and 2D-NMR.

RESULTS: The extracts inhibited the growth of MtbH37Rv [minimum inhibitory concentration (MIC) 4-64 µg/mL]. Spectroscopic characterization led to the identification of two compounds, hydroxymyristic acid methylester (1) and a substituted pyrenyl ester, a sterol (2). Compounds 1 and 2 had MIC of 49.45 and 46.99 µg/mL; IC(50) >100 and 38.21 µg/mL; selectivity index (SI) >2.02 and 0.81, respectively, and a minimum bactericidal concentration (MBC) of 62.50 µg/mL.

DISCUSSION AND CONCLUSIONS: The anti-TB activities of these plants on M. tuberculosis H37Rv support their use in traditional medicine for the treatment of coughs associated with TB and reveals the presence of anti-Mtb active compounds in the plants. These findings not only demonstrate a new potential area of therapeutic value of E. camaldulensis and E. torelliana, but also illustrate the role of esters as anti-Mtb active principles in ethnobotanical preparations and as lead compounds in the development of new and effective anti-Mtb drugs.

Abstract  OBJECTIVE: To determine the adulticidal and repellent activities of different solvent leaf extracts of Eclipta alba (E. alba) and Andrographis paniculata (A. paniculata) against malarial vector, Anopheles stephensi (An. stephensi).

METHODS: Adulticidal efficacy of the crude leaf extracts of E. alba and A. paniculata with five different solvents like benzene, hexane, ethyl acetate, methanol and chloroform was tested against the five to six day old adult female mosquitoes of An. stephensi. The adult mortality was observed after 24 h under the laboratory conditions. The repellent efficacy was determined against An. stephensi mosquito species at three concentrations viz., 1.0, 2.5 and 5.0 mg/cm(2) under laboratory conditions.

RESULTS: Among the tested solvents the maximum efficacy was observed in the methanol extract. The LC(50) and LC(90) values of E. alba and A. paniculata against adults of An. stephensi were 150.36, 130.19 ppm and 285.22, 244.16 ppm, respectively. No mortality was observed in controls. The chi-square values were significant at P<0.05 level. Methanol extract of E. alba and A. paniculata was produce maximum repellency against An. stephensi.

CONCLUSIONS: From the results it can be concluded the crude extract of E. alba and A. paniculata was an excellent potential for controlling An. stephensi mosquitoes.