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1.  Antifertility activity of Cryptolepis sanguinolenta leaf ethanolic extract in male rats 
Complementary medicine has grown over time with more botanicals emerging and remaining integral parts of medicare. Such botanicals include Cryptolepis sanguinolenta.
This study investigated the effect of Cryptolepis sanguinolenta leaf ethanolic extract on male reproductive system using rat model.
Control and treated rats were maintained on control diet. Treated rats also received graded doses of the extract.
When compared with the controls, Cryptolepis sanguinolenta treatment led to significant testosterone suppression associated with consequent significant rise in luteinizing hormone (LH) and decrease in sperm count. Treatment with Cryptolepis sanguinolenta did not result in significant attenuation of follicular stimulating hormone (FSH) levels and testicular morphometry. Sperm viability, motility, and morphology were also comparable in all groups.
These results suggest that Cryptolepis sanguinolenta possesses anti-androgenic and anti-spermatogenic properties with potential anti-aphrodisiac activity.
PMCID: PMC3409919  PMID: 22870014
Cryptolepis sanguinolenta; FSH; LH; sperm; testes; testosterone
2.  Effect of ethanolic extract of Cryptolepis sanguinolenta stem on in vivo and in vitro glucose absorption and transport: Mechanism of its antidiabetic activity 
Extracts from various morphological parts of Cryptolepis sanguinolenta are widely used traditionally in folklore medicine in many parts of the world for the management, control, and/or treatment of a plethora of human ailments, including diabetes mellitus. In order to scientifically appraise some of the ethnomedical uses of Cryptolepis sanguinolenta, the present study was undertaken to investigate its influence at varying doses on intestinal glucose absorption and transport in relation to its hypoglycemic and hypolipidemic effects in rat experimental paradigms.
Materials and Methods:
The animals used were divided into four groups. Control animals received 2 ml of distilled water, while treated groups received 50, 150, and 250 mg/kg bw of Cryptolepis sanguinolenta extract per oral respectively daily for 21 days.
Cryptolepis sanguinolenta led to a significant decrease in glucose transport and absorption. It also caused significant reductions in plasma glucose, total cholesterol, triglyceride, and LDL cholesterol. Biochemical changes observed were suggestive of dose dependence. Histopathological studies also showed increased sizes of β cells of the pancreas.
The findings in these normoglycemic laboratory animals suggest that Cryptolepis sanguinolenta has hypoglycemic and hypolipidemic activities, possibly by reducing glucose absorption and transport, and enhancing the structural and functional abilities of the β cells. This is the first study to report the effect of Cryptolepis sanguinolenta on intestinal glucose absorption. This effect could be attributed to its major bioactive principle, cryptolepine, an indoloquinoline alkaloid. This study thus lends credence to the use of Cryptolepis sanguinolenta in the management of diabetes mellitus.
PMCID: PMC3354926  PMID: 22701855
β cells; Cryptolepis sanguinolenta; diabetes; glucose; lipid
3.  A Review of the Anticancer Potential of The Antimalarial Herbal Cryptolepis Sanguinolenta and Its Major Alkaloid Cryptolepine 
Ghana Medical Journal  2013;47(3):137-147.
Cryptolepis sanguinolenta (Lindl.) Schltr (Periplocaceae), has a longstanding traditional use in the treatment of malaria in the West African region. Recent evidence suggests that the aqueous extract from the roots and the major alkaloid from the plant, cryptolepine, have prospects as cancer chemotherapeutic agents on account of their potent cytotoxicity to mammalian cells. Several mechanisms have been proposed to explain the cytotoxic activities of the agents. However, emerging evidence from their anti-inflammatory actions suggest that the mechanism of the cytotoxicity may be closely related to its anti-inflammatory activity. This review looks at the mechanisms of cryptolepis-induced cytotoxicity, its link with inflammation and its potential as anticancer agent. The elucidation of these interwoven mechanisms may be useful in the development of cryptolepine or other analogues as new anticancer agents.
PMCID: PMC3875281  PMID: 24391229
4.  Clinical Efficacy of a Tea-Bag Formulation of Cryptolepis Sanguinolenta Root in the Treatment of Acute Uncomplicated Falciparum Malaria 
Ghana Medical Journal  2010;44(1):3-9.
To evaluate the clinical efficacy and safety of a tea bag formulation of the root of C. sanguinolenta
This is a prospective descriptive open trial.
Patients were recruited from Korle-Bu, Mamprobi and Dansoman Polyclinics.
Forty-four subjects with uncomplicated malaria were recruited for the study.
Patients presented with clinical symptoms of malaria. Laboratory investigations conducted included malaria parasite counts, haematological indices and biochemical tests which were obtained before, during and after a 5-day treatment period, up to Day 28 posttreatment. All patients in the study were symptomatic with significant parasitaemia. Patients were given one teabag three times a day, that is, morning, noon and night, for five days of treatment.
Fifty percent of the patients were cleared of their P. falciparum parasitaemia by 72 hours, and all by Day 7. Presenting symptoms of fever, chills, nausea and vomiting cleared rapidly, all by Day 3, but resolution of haematological and biochemical abnormalities associated with malaria was generally slow, a feature seen in malaria post-treatment. The overall cure rate was 93.5% due to two cases of recrudescence on Days 21 and 28. The laboratory findings did not suggest any toxicity.
On the basis of fever clearance and disappearance of parasitaemia by Day 7, the formulation has been shown to be non-toxic and highly effective in the treatment of acute uncomplicated malaria.
PMCID: PMC2956309  PMID: 21326984
Clinical efficacy; Cryptolepis sanguinolenta; falciparum malaria; Ghana; tea bag formulation
5.  Antimicrobial activity and phytochemical analyses of Polygonum aviculare L. (Polygonaceae), naturally growing in Egypt 
Polygonum aviculare (Polygonaceae) is an herb commonly distributed in Mediterranean coastal regions in Egypt and used in folkloric medicine. Organic and aqueous solvent extracts and fractions of P. aviculare were investigated for antimicrobial activities on several microorganisms including bacteria and fungi. Phytochemical constituents of air-dried powered plant parts were extracted using aqueous and organic solvents (acetone, ethanol, chloroform and water). Antimicrobial activity of the concentrated extracts was evaluated by determination of the diameter of inhibition zone against both Gram-negative and Gram-positive bacteria and fungi using paper disc diffusion method.
Results of the phytochemical studies revealed the presence of tannins, saponins, flavonoids, alkaloids and sesquiterpenes and the extracts were active against both Gram-negative and Gram-positive bacteria. Chloroform extract gave very good and excellent antimicrobial activity against all tested bacteria and good activity against all tested fungi except Candida albicans. Structural spectroscopic analysis that was carried out on the active substances in the chloroform extract led to the identification of panicudine (6-hydroxy-11-deoxy-13 dehydrohetisane).
Evaluation of the antimicrobial activity of panicudine indicated significant activity against all tested Gram-negative and Gram-positive organisms. Panicudine displayed considerable activity against the tested fungi with the exception of C. albicans. Antimicrobial activity of the extracts was unaffected after exposure to different heat treatments, but was reduced at alkaline pH. Studies of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of panicudine on the tested organisms showed that the lowest MIC and the MBC were demonstrated against Salmonella paratyphi, Bacillus subtilis and Salmonella typhi and the highest MIC and MBC were against Staphylococcus aureus.
PMCID: PMC3730555  PMID: 23961059
Polygonum aviculare; Antimicrobial activity; Phytochemical analysis; Minimum inhibitory concentration; Minimum bactericidal concentration
8.  Antimicrobial activities of the rhizome extract of Zingiber zerumbet Linn 
To investigate antimicrobial effects of ethanolic extract of Zingiber zerumbet (Z. zerumbet) (L.) Smith and its chloroform and petroleum ether soluble fractions against pathogenic bacteria and fungi.
The fresh rhizomes of Zingiber zerumbet were extracted in cold with ethanol (4.0 L) after concentration. The crude ethanol extract was fractionated by petroleum ether and chloroform to form a suspension of ethanol extract (15.0 g), petroleum ether fraction (6.6 g) and chloroform soluble fraction (5.0 g). The crude ethanol extract and its petroleum ether and chloroform fractions were evaluated for antibacterial and antifungal activity against thirteen pathogenic bacteria and three fungi by the disc diffusion method. Commercially available kanamycin (30 µg/disc) was used as standard disc and blank discs impregnated with the respective solvents were used as negative control.
At a concentration of 400 µg/disc, all the samples showed mild to moderate antibacterial and antifungal activity and produced the zone of inhibition ranging from 6 mm to 10 mm. Among the tested samples, the crude ethanol extract showed the highest activity against Vibrio parahemolyticus (V. parahemolyticus). The minimum inhibitory concentration (MIC) of the crude ethanol extract and its fractions were within the value of 128-256 µg/mL against two Gram positive and four Gram negative bacteria and all the samples showed the lowest MIC value against V. parahemolyticus (128 µg/mL).
It can be concluded that, potent antibacterial and antifungal phytochemicals are present in ethanol extract of Z. zerumbet (L).
PMCID: PMC3614197  PMID: 23569803
Zingiber zerumbet (L.) Smith; Antibacterial; Antifungal; Minimum inhibitory concentration (MIC); Antimicrobial activity; Rhizome; Antifungal activity; Phytochemicals; Disc diffusion method; Soluble fraction; Pathogenic bacteria
9.  Antimicrobial Efficacy and Phytochemical Analysis of Indigofera Trita Linn. 
An in vitro antimicrobial activity and phytochemical analysis of various extracts of Indigofera trita L. viz. petroleum ether, chloroform, acetone, ethanol and aqueous extracts were carried out. A total of 21 microorganisms (19 bacteria and 2 fungal strains) were used for antimicrobial activity by disc diffusion method and a standard procedure was used to identify the phytochemical constituents. Petroleum ether extract showed moderate inhibitory activity against Staphylococcus aureus (14.40 mm), S. epidermidis (14.20 mm), Salmonella paratyphi A (12.80 mm), Streptococcus mutans (12.20 mm), Escherichia coli, Proteus vulgaris, S. typhi and Burkholderia cepacia (12.00 mm). The chloroform extract also showed antimicrobial activity against S. epidermidis (14.20 mm), S. typhimurium (12.60 mm), S. paratyphi A, S. brunei and Yersinia enterocolitica (12.00 mm). The acetone extract of I. trita showed considerable inhibitory activity against S. epidermidis (18.20 mm), S. typhimurium (14.60 mm), S. infantis (13.80 mm), S. aureus (13.40 mm), Y. enterocolitica (13.00 mm) and Enterobacter aerogenes (12.00 mm) were documented. Ethanol extract showed significant antimicrobial activity against S. epidermidis (18.60 mm), S. paratyphi A (14.60 mm), Y. enterocolitica (13.40 mm), S. typhi (12.40 mm), S. aureus, E. aerogenes, S. typhimurium and S. infantis (12.00 mm). Aqueous extract of I. trita considerably inhibited S. epidermidis (13.80 mm), S. paratyphi A and Y. enterocolitica (12.20 mm), E. aerogenes and Haemophilus parahaemolyticus (12.00 mm). All the five extracts showed a minimal antifungal activity when compared to antibacterial activity. The result revealed that the antimicrobial properties of I. trita might be associated with the presence of phenolic compounds, flavonoids, tannins, glycosides, saponins, phytosterols and alkaloids.
PMCID: PMC3777595  PMID: 24146483
Inidigofera trita; Phytoconstituents; Antimicrobial activity; Antifungal activity; Disc diffusion method
10.  Bauhinia variegata Leaf Extracts Exhibit Considerable Antibacterial, Antioxidant, and Anticancer Activities 
BioMed Research International  2013;2013:915436.
The present study reports the phytochemical profiling, antimicrobial, antioxidant, and anticancer activities of Bauhinia variegata leaf extracts. The reducing sugar, anthraquinone, and saponins were observed in polar extracts, while terpenoids and alkaloids were present in nonpolar and ethanol extracts. Total flavonoid contents in various extracts were found in the range of 11–222.67 mg QE/g. In disc diffusion assays, petroleum ether and chloroform fractions exhibited considerable inhibition against Klebsiella pneumoniae. Several other extracts also showed antibacterial activity against pathogenic strains of E. coli, Proteus spp. and Pseudomonas spp. Minimum bactericidal concentration (MBC) values of potential extracts were found between 3.5 and 28.40 mg/mL. The lowest MBC (3.5 mg/mL) was recorded for ethanol extract against Pseudomonas spp. The antioxidant activity of the extracts was compared with standard antioxidants. Dose dependent response was observed in reducing power of extracts. Polar extracts demonstrated appreciable metal ion chelating activity at lower concentrations (10–40 μg/mL). Many extracts showed significant antioxidant response in beta carotene bleaching assay. AQ fraction of B. variegata showed pronounced cytotoxic effect against DU-145, HOP-62, IGR-OV-1, MCF-7, and THP-1 human cancer cell lines with 90–99% cell growth inhibitory activity. Ethyl acetate fraction also produced considerable cytotoxicity against MCF-7 and THP-1 cell lines. The study demonstrates notable antibacterial, antioxidant, and anticancer activities in B. variegata leaf extracts.
PMCID: PMC3777169  PMID: 24093108
11.  Crystallization and preliminary X-ray analysis of cryptolepain, a novel glycosylated serine protease from Cryptolepis buchanani  
Cryptolepain is a stable glycosylated novel serine protease was crystallized by hanging-drop method. Crystal data was processed up to 2.25 Å with acceptable statistics and structure determination of the enzyme is under way.
Cryptolepain is a stable glycosylated novel serine protease purified from the latex of the medicinally important plant Cryptolepis buchanani. The molecular weight of the enzyme is 50.5 kDa, as determined by mass spectrometry. The sequence of the first 15 N-terminal resides of the protease showed little homology with those of other plant serine proteases, suggesting it to be structurally unique. Thus, it is of interest to solve the structure of the enzyme in order to better understand its structure–function relationship. X-ray diffraction data were collected from a crystal of cryptolepain and processed to 2.25 Å with acceptable statistics. The crystals belong to the orthorhombic space group C2221, with unit-cell parameters a = 81.78, b = 108.15, c = 119.86 Å. The Matthews coefficient was 2.62 Å3 Da−1 with one molecule in the asymmetric unit. The solvent content was found to be 53%. Structure determination of the enzyme is under way.
PMCID: PMC2330135  PMID: 17277443
cryptolepain; serine proteases
12.  Changes in Carbohydrate and Nutrient Contents Throughout a Reproductive Cycle Indicate that Phosphorus is a Limiting Nutrient in the Epiphytic Bromeliad, Werauhia sanguinolenta 
Annals of Botany  2006;97(5):745-754.
• Background and Aims This study examined the physiological basis of the cost of reproduction in the epiphytic bromeliad Werauhia sanguinolenta, growing in situ in a tropical lowland forest in Panama.
• Methods Entire mature plants were sampled repeatedly over the course of 2 years, which represents the common interval between reproductive events. Due to the uncertainty concerning the appropriate currency of resource allocation to reproduction, the temporal changes of the contents of total non-structural carbohydrates (TNC) and of all major nutrient elements in different plant parts were studied (stems, green leaves, non-green leaf bases, roots and reproductive structures when present).
• Key Results Although TNC varied with time in all compartments, this variation was more related to seasonal fluctuations than to reproductive status. The contents of the nutrient elements, N, P, K, Mg and S, on the other hand, showed significant differences between reproductive and non-reproductive individuals, while Ca did not change with reproductive status. Differences in nutrient contents were most pronounced in stems. Seeds were particularly enriched in P, much less so in N and the other nutrient elements. Model calculations of nutrient fluxes indicate that a plant needs about 2 years to accumulate the amount of P invested in a fruit crop, while the estimated uptake rates for N were much faster.
• Conclusions Since most mature individuals of this species fruit every other year, it is hypothesized that P is the prime limiting factor for reproduction. These findings therefore add to an increasing body of evidence that P rather than N is limiting growth and reproduction in vascular epiphytes.
PMCID: PMC2803411  PMID: 16497701
Barro Colorado Island; Bromeliaceae; cost of reproduction; nitrogen; non-structural carbohydrates; nutrient uptake; phosphorus; reproductive investment; seeds; Werauhia sanguinolenta
13.  Tagetes erecta Linn. and its mosquitocidal potency against Culex quinquefasciatus 
To investigate mosquitocidal effects of ethanolic extract of flowers of Tagetes erecta (T. erecta) and its chloroform and petroleum ether soluble fractions against the larvae of Culex quinquefasciatus (Cx. quinquefasciatus).
The fresh flowers of T. erecta were extracted in cold with ethanol (5.0 L) and after concentration, the ethanol extract was fractionated with chloroform and petroleum ether to afford a brownish syrupy suspension of ethanol extract (50.0 g), petroleum ether soluble fraction (18.6 g) and chloroform soluble fraction (23.8 g). The larvicidal effect of ethanol extract and their solvent fractions were determined by the standard procedure of WHO against different instars of Cx. quinquefasciatus.
Among the tested samples the chloroform soluble fractions showed the highest toxicity and consequently, the lowest LC50 values (14.14 µg/mL, 17.06 µg/mL, 36.88 µg/mL and 75.48 µg/mL) for all the instars larvae of Cx. quinquefasciatus. The larvae showed comparative tolerance in the course of increasing age and time.
It can be concluded that the flowers of T. erecta are very effective natural larvicide and could be useful against Cx. quinquefasciatus.
PMCID: PMC3609190  PMID: 23569756
Tagetes erecta Linn.; Culex quinquefasciatus Say; Mosquitocidal; Larvae; Larvicidal effect; Toxicity
Ancient Science of Life  1992;12(1-2):251-256.
The ethanolic extract of roots of Salacia macrosperma (Hippocrataceae) exhibited significant in vitro antimicrobial activity. The ethanolic extract was fractionated with different solvents and all the fractions were screened for their antimicrobial spectrum against eight gram-positive, five gram-negative and ten fungal strains. Chloroform fraction followed by benzene fraction of enthanolic extract showed significant antimicrobial effect against all the microorganisms tested. The dose-dependent activity of fractions was evaluated against selected microbial strains and compared with appropriate standards.
PMCID: PMC3336611  PMID: 22556596
15.  Comparative antimicrobial activity of callus and natural plant extracts of Solanum trilobatum L 
Ancient Science of Life  2009;28(3):3-5.
Comparison of natural plant and callus extracts of Solanum trilobatum L. was studied against two bacteria and fungi, for their antimicrobial activity using cup diffusion method. Various solvents such as chloroform, petroleum ether and ethanol were used. The leaf and stem segments of the plant were culturedon Murashige and S koog basal medium supplemented with various growth regulators. Maximum callus was recorded on medium containing 0.5 mg/lNAA and 0.5 mgj IKinetin. The results reveals that the stem and leaf callus extracts has shown significant activity against the tested microorganisms than the natural sample.
PMCID: PMC3336318  PMID: 22557312
Solanum trilobatum; callus; kinetin; extracts; antimicrobial activity
16.  Antimicrobial potential of Ricinus communis leaf extracts in different solvents against pathogenic bacterial and fungal strains 
To investigate the in vitro antimicrobial activities of the leaf extract in different solvents viz., methanol, ethanol and water extracts of the selected plant Ricinus communis.
Agar well diffusion method and agar tube dilution method were carried out to perform the antibacterial and antifungal activity of methanol, ethanol and aqueous extracts.
Methanol leaf extracts were found to be more active against Gram positive bacteria (Bacillus subtilis: ATCC 6059 and Staphylococcus aureus: ATCC 6538) as well as Gram negative bacteria (Pseudomonas aeruginosa: ATCC 7221 and Klebsiella pneumoniae) than ethanol and aqueous leaf extracts. Antifungal activity of methanol and aqueous leaf extracts were also carried out against selected fungal strains as Aspergillus fumigatus and Aspergillus flavus. Methanolic as well as aqueous leaf extracts of Ricinus communis were effective in inhibiting the fungal growth.
The efficient antibacterial and antifungal activity of Ricinus communis from the present investigation revealed that the methanol leaf extracts of the selected plant have significant potential to inhibit the growth of pathogenic bacterial and fungal strains than ethanol and aqueous leaf extracts.
PMCID: PMC3621469  PMID: 23593573
Antibacterial; Antifungal; Relative percentage inhibition; Ricinus communis; Methanol; Ethanol
17.  Antibacterial activities and phytochemical analysis of Cassia fistula (Linn.) leaf 
Cassia fistula Linn. which belongs to family Leguminosae is a medium-sized tree and its different parts are used in ayurvedic medicine as well as home remedies for common ailments. Sequential extraction was carried out using solvents viz. petroleum ether, chloroform, ethanol, methanol and water from leaf of the plant were investigated for preliminary phytochemical and antibacterial property. Results of the study showed that all the extracts had good inhibitory activity against Gram-positive test organism. Although all five extracts showed promising antibacterial activity against test bacterial species, yet maximum activity was observed in ethanol extract. The minimum inhibitory concentration ranged in between 94 to 1 500 μg/ml. Evaluation of phytochemicals such as alkaloids, flavonoids, carbohydrates, glycosides, protein and amino acids, saponins, and triterpenoids revealed the presence of most of constituents in polar extracts (ethanol, methanol, and aqueous) compared with nonpolar extracts (petroleum ether and chloroform). Furthermore, the ethanol extract was subjected to TLC bioautography and time-kill study against Staphylococcus epidermidis. All the findings exhibit that the leaf extracts have broad-spectrum activity and suggest its possible use in treatment of infectious diseases.
PMCID: PMC3217677  PMID: 22171295
Cassia fistula; human pathogenic bacteria; minimum inhibitory concentration; Similipal Biosphere Reserve; TLC bioautography
18.  Blechnum Orientale Linn - a fern with potential as antioxidant, anticancer and antibacterial agent 
Blechnum orientale Linn. (Blechnaceae) is used ethnomedicinally for the treatment of various skin diseases, stomach pain, urinary bladder complaints and sterilization of women. The aim of the study was to evaluate antioxidant, anticancer and antibacterial activity of five solvent fractions obtained from the methanol extract of the leaves of Blechnum orientale Linn.
Five solvent fractions were obtained from the methanol extract of B. orientale through successive partitioning with petroleum ether, chloroform, ethyl acetate, butanol and water. Total phenolic content was assessed using Folin-Ciocalteu's method. The antioxidant activity was determined by measuring the scavenging activity of DPPH radicals. Cytotoxic activity was tested against four cancer cell lines and a non-malignant cell using MTT assay. Antibacterial activity was assessed using the disc diffusion and broth microdilution assays. Standard phytochemical screening tests for saponins, tannins, terpenoids, flavonoids and alkaloids were also conducted.
The ethyl acetate, butanol and water fractions possessed strong radical scavenging activity (IC50 8.6-13.0 μg/ml) and cytotoxic activity towards human colon cancer cell HT-29 (IC50 27.5-42.8 μg/ml). The three extracts were also effective against all Gram-positive bacteria tested: Bacillus cereus, Micrococcus luteus, methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA) and Stapylococcus epidermidis(minimum inhibitory concentration MIC 15.6-250 μg/ml; minimum bactericidal concentration MBC 15.6-250 μg/ml). Phytochemical analysis revealed the presence of flavonoids, terpenoids and tannins. Ethyl acetate and butanol fractions showed highest total phenolic content (675-804 mg gallic acid equivalent/g).
The results indicate that this fern is a potential candidate to be used as an antioxidant agent, for colon cancer therapy and for treatment of MRSA infections and other MSSA/Gram-positive bacterial infectious diseases.
PMCID: PMC2881084  PMID: 20429956
19.  Antibacterial Screening of Aegle Marmelos, Lawsonia Inermis and Albizzia Libbeck 
Three medicinal plant Aegle marmelos, Lawsonia inermis, Albizzia libbeck were extracted by soxhlet apparatus using petroleum ether, ethanol, chloroform and aqueous as solvent. Among those extract, the petroleum ether was considered as effective one. The extracts were subjected to preliminary phytochemical screening and the three plants with four extracts were tested against three Gram positive bacteria (B.cereus, B.subtilis, S. aureus) and three Gram negative bacteria (E.coli, P.vulgaris, and P.aeruginosa) by disc diffusion method. Maximum inhibition (3.8cm) was recorded in Lawsonia inermis. It also showed inhibitory action against all the six pathogen tested. The zone of inhibition of the extracts was compared with the standard antibiotics Streptomycin and Spectinomycin. The study suggests that the plant is promising the development of phytomedicine for antimicrobial properties.
PMCID: PMC2816442  PMID: 20162092
Antibacterial activity; Aegle marmelos; Lawsonia inermis; Albizzia libbeck
20.  Bacteriocin from Bacillus subtilis as a novel drug against diabetic foot ulcer bacterial pathogens 
To isolate and identify Bacillus subtilis (B. subtilis) from soil and to characterize and partially purify the bacteriocin. To evaluate the antimicrobial activity against four diabetic foot ulcer bacterial pathogens.
Genotypic identification was done based on Bergey's manual of systemic bacteriology. Antimicrobial susceptibility test was done by Kirby-Bauer disc diffusion method. Colonies were identified by colony morphology and biochemical characterization and also compared with MTCC 121 strain. Further identification was done by 16S rRNA sequencing. Inhibitory activities of partially purified bacteriocin on all the DFU isolates were done by agar well diffusion method. The strain was identified to produce bacteriocin by stab overlay assay. Bacteriocin was extracted by organic solvent extraction using chloroform, further purified by HPLC and physical, and chemical characterization was performed.
The four isolates showed high level of resistance to amoxyclav and sensitivity to ciprofloxacin. HPLC purification revealed that the extracts are bacteriocin. The phylogenetic tree analysis results showed that the isolate was 99% related to B. subtilis BSF01. The results reveled activity to all the four isolates and high level of activity was seen in case of Klebsiella sp.
Partially purified bacteriocin was found to have antimicrobial activity against the four diabetic foot ulcer bacterial pathogens, which can thus be applied as a better drug molecule on further studies. The strain B. subtilis are found to be safe for use and these antimicrobial peptides can be used as an antimicrobial in humans to treat DFU bacterial pathogens.
PMCID: PMC3805101  PMID: 24093784
16S rRNA; Antimicrobial activity; Micrococcus leuteus; HPLC; Physicochemical characterization
21.  Antimicrobial Property of Extracts of Indian Lichen against Human Pathogenic Bacteria 
Context. Usnea ghattensis G. Awasthi (Usneaceae) endemic fruticose lichen found growing luxuriantly in Northern Western Ghats of India, it also contains Usnic acid as a major chemical and tested against some human pathogenic bacteria. Objective. To explore antimicrobial properties of Usnea ghattensis against some human pathogenic bacteria. Materials and Methods. The lichen was extracted in acetone, methanol, and ethanol. In vitro antimicrobial activity was tested initially by Kirby-Bauer technique of disc diffusion method and was confirmed by minimum inhibitory concentration using Broth microdilution method according to the NCCLS guidelines. Results. Ethanol extract was most effective against Bacillus cereus and Pseudomonas aeruginosa with a zone of inhibition 29.8 ± 0.6 mm and 12.3 ± 0.5 mm diameters at a concentration of 0.2 mg/mL. Acetone and methanol extract demonstrated almost similar activity against Staphylococcus aureus and the zone of inhibition was 24.6 ± 0.5 and 24.7 ± 0.4 mm. Only methanol extract was showing activity against Streptococcus faecalis with a 13.5 ± 0.8 mm zone. MIC value noted against Staphylococcus aureus and Streptococcus faecalis was 6.25 μg/mL and 25 μg/mL, whereas against Bacillus cereus and Pseudomonas aeruginosa, MIC calculated was 3.125 μg/mL and 200 μg/mL, respectively. Conclusion. The present study demonstrates the relatively higher activity of this lichen against not only gram (+) but significantly also against gram (−) bacteria. This indicates that this lichen might be a rich source of effective antimicrobial agents.
PMCID: PMC3767054  PMID: 24062769
22.  Preliminary phytochemical and antibacterial screening of Sesuvium portulacastrum in the United Arab Emirates 
Pharmacognosy Research  2012;4(4):219-224.
The present study describes the phytochemical profile and antimicrobial activity of Sesuvium portulacastrum.
Materials and Methods:
Three extracts of S. portulacastrum obtained by extraction in aqueous, ethanolic and dichloromethane solvents, respectively, were compared for their antimicrobial activity and ethanolic extract further subjected to gas chromatography-mass spectrometry (GC-MS) analysis to find out the nature of the compounds responsible for the antimicrobial activity. The antibacterial activities were assessed by measuring the diameter of the inhibition zones, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values.
Compared to the aqueous and dichloromethane extract, the ethanolic extract showed better antimicrobial activity against Staphylococcus aureus and E. coli, indicating its potential application related to noscomial infections. GC-MS results revealed 22, 23-Dihydrostigmasterol, Benzoic acid, 3,4,5-trihydroxy-(Gallic acid), (2R,3R)-(-)-Epicatechin and Capsaicin in the ethanolic extract to be the molecules responsible for the antimicrobial activity of S. portulacastrum.
To the best of our knowledge, this is the first report on analysis of antimicrobial components from S. portulacastrum in United Arab Emirates (UAE), and our results confer the utility of this plant extract in developing a novel broad spectrum antimicrobial agent.
PMCID: PMC3510875  PMID: 23225966
Antibacterial; gas chromatography-mass spectrometry; phytochemical; Sesuvium portulacastrum
23.  Assessment of phytochemicals, antimicrobial and cytotoxic activities of extract and fractions from Fagonia olivieri (Zygophyllaceae) 
In Pakistan Fagonia olivieri (Zygophyllaceae) is commonly used in the indigenous system of medicine for treatment of conditions like diabetes, cancer, fever, asthma, toothache, stomach troubles and kidney disorders. This study evaluated the crude methanol extract of F. olivieri (FOM) and its derived fractions for their antimicrobial and cytotoxic activities as well as the classes of phytochemical.
Dried powder of whole plant of F. olivieri was extracted with methanol (FOM) and the resultant was fractionated to give n-hexane fraction (FOH), chloroform fraction (FOC), ethyl acetate fraction (FOE), n-butanol fraction (FOB) and residual aqueous fraction (FOA). Methanol extract and its derived fractions were subjected to phytochemical screening using standard procedures. Also the extract and fractions were assayed for antibacterial, antifungal and cytotoxic activities using agar well diffusion technique, agar tube dilution method and brine shrimps lethality test, respectively.
The results obtained for phytochemical analysis indicate the presence of saponins and alkaloids in all the tested extract and fractions while anthraquinones were not detected. The results showed that all the bacterial strains tested in this study were susceptible to at least one of the fractions tested. However, FOE and FOB were the best antibacterial fractions and showed antibacterial activity against maximum number of bacterial strains. The results showed that Escherichia coli was the most sensitive bacterium while Bordetella bronchiseptica and Enterobacter aerogenes were less susceptible against various fractions. Maximum percent inhibition for growth was recorded for the fungus Aspergillus flavus with FOE whereas growth of Aspergillus fumigatus and Fusarium solani was inhibited by FOM and its all derived fractions. Minimum LC50 (24.07 mg/L) for brine shrimp assay was recorded for FOE followed by LC50 of FOC (26.1 mg/L) and FOB (30.05 mg/L) whereas maximum LC50 was exhibited by FOH (1533 mg/L).
These results indicated the use of F. olivieri to treat infections with emphasis to isolate and characterize the active principle responsible for antibacterial, antifungal and cytotoxic activities and its exploitation as therapeutic agent.
PMCID: PMC3717109  PMID: 23842440
Antibacterial activity; Brine shrimps lethality assay; Cytotoxicity; Fagonia olivieri; Phytochemicals
24.  Anticancer property of Bryophyllum pinnata (Lam.) Oken. leaf on human cervical cancer cells 
Bryophyllum pinnata (B. pinnata) is a common medicinal plant used in traditional medicine of India and of other countries for curing various infections, bowel diseases, healing wounds and other ailments. However, its anticancer properties are poorly defined. In view of broad spectrum therapeutic potential of B. pinnata we designed a study to examine anti-cancer and anti-Human Papillomavirus (HPV) activities in its leaf extracts and tried to isolate its active principle.
A chloroform extract derived from a bulk of botanically well-characterized pulverized B. pinnata leaves was separated using column chromatography with step- gradient of petroleum ether and ethyl acetate. Fractions were characterized for phyto-chemical compounds by TLC, HPTLC and NMR and Biological activity of the fractions were examined by MTT-based cell viability assay, Electrophoretic Mobility Shift Assay, Northern blotting and assay of apoptosis related proteins by immunoblotting in human cervical cancer cells.
Results showed presence of growth inhibitory activity in the crude leaf extracts with IC50 at 552 μg/ml which resolved to fraction F4 (Petroleum Ether: Ethyl Acetate:: 50:50) and showed IC50 at 91 μg/ml. Investigations of anti-viral activity of the extract and its fraction revealed a specific anti-HPV activity on cervical cancer cells as evidenced by downregulation of constitutively active AP1 specific DNA binding activity and suppression of oncogenic c-Fos and c-Jun expression which was accompanied by inhibition of HPV18 transcription. In addition to inhibiting growth, fraction F4 strongly induced apoptosis as evidenced by an increased expression of the pro-apoptotic protein Bax, suppression of the anti-apoptotic molecules Bcl-2, and activation of caspase-3 and cleavage of PARP-1. Phytochemical analysis of fraction F4 by HPTLC and NMR indicated presence of activity that resembled Bryophyllin A.
Our study therefore demonstrates presence of anticancer and anti-HPV an activity in B. pinnata leaves that can be further exploited as a potential anticancer, anti-HPV therapeutic for treatment of HPV infection and cervical cancer.
PMCID: PMC3353166  PMID: 22405256
Bryophyllum pinnata; Human Papillomavirus; HeLa cells; AP1; NMR; HPTLC
25.  Antibacterial Activity of some Medicinal Mangroves against Antibiotic Resistant Pathogenic Bacteria 
The antibacterial activity of the leaves and bark of mangrove plants, Avicennia marina, A. officinalis, Bruguiera sexangula, Exoecaria agallocha, Lumnitzera racemosa, and Rhizophora apiculata was evaluated against antibiotic resistant pathogenic bacteria, Staphylococcus aureus and Proteus sp. Soxhlet extracts of petroleum ether, ethyl acetate, ethanol and water were prepared and evaluated the antibacterial activity using agar diffusion method. Most of the plant extracts showed promising antibacterial activity against both bacterial species. However, higher antibacterial activity was observed for Staphylococcus aureus than Proteus sp. The highest antibacterial activity was shown by ethyl acetate of mature leaf extracts of E. agallocha for Staphylococcus aureus. All ethyl acetate extracts showed higher inhibition against S. aureus while some extracts of chloroform, ethyl acetate and ethanol gave inhibition against Proteus sp. None of the petroleum ether and aqueous extracts showed inhibition against Proteus sp. All fresh plant materials did also show more antibacterial activity against both bacterial strains than did dried plant extracts. Antibacterial activity of fresh and dried plant materials reduced for both bacterial strains with time after extraction. Since L. racemosa and A. marina gave the best inhibition for bacterial species, they were used for further investigations. Charcoal treated plant extracts of L. racemosa and A. marina were able to inhibit both bacterial strains more than those of untreated plant extracts. Phytochemical screening of mature leaf, bark of L. racemosa and leaf extracts of A. marina has been carried out and revealed that leaf and bark contained alkaloids, steroids, triterpenoids and flavonoids. None of the above extracts indicate the presence of saponins and cardiac glycosides. Separated bands of extracts by TLC analysis showed antibacterial activity against S. aureus.
PMCID: PMC2929774  PMID: 20838519
Antibacterial activity; inhibition; mangroves; soxhlet extraction

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