Application of microscopy and spectroscopy in investigating anti-cancer potential of Achyranthes aspera L. leaves.

The genus Achyranthes belong to the family Amaranthaceae which constitutes an important group of herbs and shrubs with immense medicinal value. The present research work was conducted to investigate the anticancer potential of Achyranthes aspera L. leaves by focusing on the antioxidant, aniproliferative and antimitotic activities of leaf extracts. Plant extraction was carried out by soxhelt method with different solvents. Phytochemical characterization of the plants extracts using chemical methods identified the presence of cardiac glycosides, saponins, coumarins, proteins, tannins, flavonoids and triterpenes. Alkaloid was present in methanolic and ethanolic extract. High performance liquid chromatography showed presence of different concentration of myricetin, quercetin and kaempferol in different extracts with the highest concentration of myricetin (84.53 µg/mL) in n-butanolic extract. The extracts were then tested for antioxidant activity using 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging assay by spectrophotometric method. In DPPH radical scavenging assay, antioxidant activity of A. aspera ranged between 79.78 ± 0.034% and 58.63 ± 0.069%. Highest antioxidant activity was observed for methanolic extract and lowest for acetone. Antimitotic activity was determined by using Allium cepa assay in which microscopic investigation was carried out to observe normal and abnormal phases of mitosis. In this assay, n-butanolic extract had highest antimitotic activity with minimum mitotic index at 2 mg/mL (57 ± 0.0351%). The plant extracts also caused chromosomal and mitotic aberrations which were clearly observed under 40× and 100× magnification of compound microscope. Antiproliferative activity was determined by using yeast cell model in which light microscope with hemocytometer was used for cell counting. In case of Antiproliferative activity, the ethyl acetate extract of A. aspera had highest antiproliferative activity with lowest cell viability (22.14 ± 0.076%) at highest extract concentration (2 mg/mL) while methanol extract of A. aspera had highest antiproliferative activity with lower cell viability (24.24 ± 0.057%) at lowest extract concentration (0.25 mg/mL). The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent. RESEARCH HIGHLIGHTS: Achyranthes aspera L. leaves have various phytochemicals which contribute to its medicinal properties Various extracts of the leaves of A. aspera L. possess antioxidant, antimitotic and antiproliferative potential The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent.

Phytochemical characterization and phospholipase A2 inhibitory effect of Vitex negundo L. root extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Snake bites are a critical health issue in many parts of the world particularly in Asian countries lacking efficient health facilities in rural areas. Cobra is the most common snake type in Asia and is responsible for a large number of mortalities particularly in rural areas. Plants are usually considered the most effective and easy-to-approach treatment for snake bites in rural areas of various countries. Vitex negundo L. is an important medicinal plant traditionally used to treat snake bite envenomation in many countries of Asia. AIM OF THE STUDY: From literature survey of plants traditionally used in the treatment of snake bites in Asian countries including India, Pakistan and Sri Lanka, roots of V. negundo were selected for the present study. Anti-snake venom potential of its roots was assessed through various in vitro assays targeting the phospholipase A2 enzyme. MATERIALS AND METHODS: V. negundo roots were sequentially extracted in different organic solvents to get fractions and in methanol to get total extract. The extracts were evaluated for phospholipase A2 (PLA2) inhibitory potential through inhibition of venom-induced hemolysis, ADP-induced platelet aggregation, PLA2-induced fatty acid hydrolysis and anticoagulant effect of cobra venom. Antioxidant power was determined using DPPH and superoxide radical scavenging assays. GC-MS and HPLC analysis was performed for the total methanol extract. RESULTS: Strong PLA2 inhibitory effect was observed for all the extracts. The ethyl acetate, acetone and methanol fractions significantly inhibited toxic effects of cobra venom under in vitro conditions. Radical scavenging potential of these fractions was also significantly high as compared to non-polar fractions in both DPPH and superoxide scavenging assays. Phytochemical analysis indicated high phenolic and flavonoid contents in these fractions. GC-MS and HPLC analysis of total methanol extract confirmed the presence of bis(2-ethylhexyl) phthalate, phenol, o-Guaiacol, palmitic acid-methyl ester, methyl stearate, quercetin and kaempferol in the plant. CONCLUSION: The study concluded that the roots of V. negundo, particularly their polar extracts, have strong PLA2 inhibitory effect against cobra venom confirming their traditional use to manage snake bites. The roots of this plant can be further studied for isolation of plant-based antisera.

Scanning electron microscopic screening; Can it be a taxonomic tool for identification of traditional therapeutic plants.

The limiting factor in the utilization of herbal medicine is the risk of replacement or substitution of their plant resources. Therefore, the authentication of medicinal plants before its use in herbal medicines is essentially required. Hence, the prime objective of this study was to provide some reliable morpho-anatomical and morpho-palynological tools for the identification of nine traditional therapeutic plants of district Lahore. For this study, scanning electron microscopic and light microscopic approaches has been employed. Results reported great variations in qualitative and quantitative morpho-anatomical features of the epidermal cell, stomatal complexes, trichomes, silica bodies and oil droplets. For example, Aegle marmelos L. can be distinguished from Cestrum diurnum L. based on epidermal cell shapes, as irregular cells in former and lobed cells shape in later one. Similarly unicellular, bicellular, and multicellular trichomes could be an identification basis for Cynodon dactylon L., Lantana camara L., and Pongamia pinnata L., respectively. In combination with these morpho-anatomical characters, SEM-based morpho-palynological characterization also reported diagnostic value. Both the Aegle marmelos and Lantana camara possessed similar cell shape (irregular) but can be delimited based on their pollen morphology as spheroidal pollen was reported in Aegle marmelos whereas prolate in Lantana camara. Moreover, quantitative characters also aid well for species delimitation, such as prolate pollen found in both Cestrum diurnum and Terminalia chebula Retz. but both of these varied in their P/E ratio, that is, 0.97 and 0.88, respectively. The leaf epidermal characters in correlation with morph-palynological traits showed potential for taxonomic resolution at the species level. RESEARCH HIGHLIGHTS: This study provides valuable morpho-anatomical and morpho-palynological tools for the nine traditional therapeutic plants of the district, Lahore, Pakistan. Light and scanning electron microscopic studies provided useful systematic data for the identification of studied plants. This systematic data include moprho-anatomical and morpho-palynological keys for the studied plants. This data can be used for future identification purposes.

Biological and optical properties of sol-gel derived ZnO using different percentages of silver contents.

The role of Ag dopant on ZnO thin films are studied. Ag doped ZnO thin films were deposited by sol-gel dip coating on glass substrates. The X-ray diffraction analysis shows hexagonal wurtzite with preferred orientation along the (101) plane. The crystallite size decreases from 33.40 nm to 28.37 nm with increase in silver doping percentage. Optical examination shows that the band gap decrease with an increase in the Ag doping in ZnO. The structural and optical results prove that Ag has substituted Zn in ZnO lattice. Silver doped ZnO is effective against Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) bacteria. The roughness and the surface oxygen species accelerate the bacteria killing properties of Ag doped ZnO. They have pharmacological utility as a replacement of the antibiotics, bactericide and disinfectants. The TGA study showed that the thermal stability of the Ag doped ZnO takes place between 380-435 °C.

Probe of ZrTiO2 thin films with TiO2-ZrO2 binary oxides deposited by dip coating technique.

Zirconium Titanium dioxide (TiO2: Zr) thin films were synthesized on glass substrates by the dip coating technique. The structural, optical, magnetic, photo-catalytic and antibacterial properties were explored for the films synthesized with zirconium concentration 10, 15, 20, 25 and 30 at.%. X-ray diffraction (XRD) revealed that the thin films contain anatase-rutile-brookite TiO2 and cubic-ZrO2 phases. Moreover, it was noticed that ZrO2 retarded the anatase-to-rutile transformation at high Zr doping percentage.The crystallite size of ZrO2 phase was in the range of 10.93-18.22 nm, whereas it was in the range of 13.5-21.59 nm for TiO2 phase for same Zr doping percentage. The band gap of films was in the range of 2.99-3.17 eV. The band gap value decreased with increase in Zr-doping percentage due to the creation localized levels near the conduction band providing a large number of electrons to reach the conduction band. Films showed room temperature ferro-magnetism due to the defects produced by Zr in TiO2 lattice. The antibacterial efficiency of ZrTiO2 thin films was investigated by utilizing disc diffusion method. Band gap was quite low, then pure TiO2 in the range 2.99-3.14 eV with Zr percentage 10-30% atom. The ZrTiO2 thin films with 20 and 25% Zr showed better antibacterial activity against Escherichia coli than 10, 15 and 30% Zr. It was found that ZrTiO2 thin films were effective solar light photo-catalyst.

Pharmacological screening of Hypericum androsaemum extracts for antioxidant, anti-lipid peroxidation, antiglycation and cytotoxicity activity.

Oxidative stress and glycation processes have a combined effect on diabetes related complications. Crude plant extracts and plant derived compounds possessing both antiglycation and antioxidant activities have a high therapeutic potential for treating these complications. Antioxidant, antiglycation, anti-lipid per oxidation and cytotoxic activities of crude methanol extract and solvent fractions of Hypericum androsaemum L. (Hypericaceae) were evaluated and correlated with total content of phenolics and flavonoids. Significant radical scavenging activity was observed for the methanol extract against 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical used as a basis for antioxidant activity with IC50 value of 92.70±2.85 µg mL(-1) (96.20±2.34% inhibition at 500 µg mL(-1)). In case of anion scavenging activity the results were not very significant (33.20±1.22% inhibition at 500 µg mL(-1)). Anti-lipid per oxidation activity was highest for n-hexane fraction (67.83±1.33% inhibition at 500 µg mL(-1)) while the ethyl acetate fraction had the highest antiglycation activity (62.77±2.54% inhibition at 500 µg mL(-1)). Statistically significant correlation was determined for antioxidant and antiglycation activity and phenolic and flavonoid contents. In cytotoxicity assay all the extracts had IC50 values >30 µg mL(-1) as compared to the standard cycloheximide (IC50 value 0.084±0.1 µg mL(-1)). The polar extracts of H. androsaemum can be a good source of non-toxic compounds with antioxidant, anti-lipid per oxidation and antiglycation activities.

Various fractions of Hypericum x moserianum and Hypericum ericoides possess antiglycation, anti-lipid peroxidation, antioxidative activities and non-toxic effects in vitro.

In the present study, two species Hypericum x moserianum and Hypericum ericoides which belong to genus Hypericum were evaluated for their potential antiglycation, antioxidant, anti lipid peroxidation and cytotoxic activities. These species are widely used in folk medicine and to the best of our knowledge there were no previous reports regarding antioxidant, anti-glycation and cytotoxicity studies of these species. Among the crude methanol extracts and fractions of both the species, the ethyl acetate fraction of H. x moserianum exhibited promising antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) with IC50 129.084±1.215µg/ml, followed by methanol extract (IC50=232.083 ± 1.215µg/ml) and aqueous fraction (IC50=266.962 ±2.213 µg/ml). The ethyl acetate fraction of H. ericoides exhibited IC50 value of 295.088 ± 2.320 µg/ml. In antiglycation assay, the ethyl acetate fraction of H. x moserianum showed 52.096% inhibition at 500µg/ml. For lipid peroxidation assay, the dichloromethane, aqueous and n-hexane fractions of H. x moserianum showed 67.241, 66.147 and 64.213% inhibition respectively, while aqueous fraction of H. ericoides exhibited 67.404% inhibition at 500µg/ml. In cytotoxicity assay, all fractions of both the species were found to be non-toxic on mouse fibroblast 3T3 cells with IC50 value greater than 30µg/ml as compared to cycloheximide with IC50 value 0.073±0.1µg/ml used as a standard. It was concluded from the study that among the two species, crude methanolic and ethyl acetate fractions were more active regarding the antioxidant, anti-glycation activities while dichloromethane, aqueous and n-hexane fractions possessed anti-lipid peroxidation activity.

A review on biological, nutraceutical and clinical aspects of French maritime pine bark extract.

Bark extract of Pinus pinaster has a long history of ethnomedicinal use and is available commercially as herbal dietary supplement with proprietary name pycnogenol. It is used as a food supplement to overcome many degenerative disorders. Rohdewald (2002) wrote the first comprehensive review of extract highlighting its antioxidative nature and its role in different diseases. Later, Watson (2003) and Gulati (2005) in their reviews about cardiovascular health, described the extract as a best neutraceutical agent in this regard. The objective of this paper is to review the current research on this extract in terms of extraction methods, its pharmacological, toxicological and nutraceutical effects and clinical studies. Web sites of Google Scholar, Pubmed and Medline were searched for articles written in English and published in peer-reviewed journals from 2006 to 2009 and sixty-nine research articles were extracted. Of these, two are about extraction advancement and analysis while the rest relate to its clinical, biological and nutraceutical aspects.

A review of the antibacterial activity of Hypericum perforatum L.

Hypericum perforatum L. (Hypericaceae) is a perennial herb that is commonly known as St. John's Wort. The plant has been valued for its important biological and chemical perspectives and its use in the treatment of infectious diseases has been documented in ethnobotanical reports. Most recent interest in H. perforatum has focused on its antidepressant effects, and only recently has its antimicrobial activity been evaluated against a number of bacterial and fungal strains. The present review gives a comprehensive summary of the ethnobotanical uses, chemical constituents and biological effects (antibacterial and antifungal) of this species. A comprehensive account of the chemical constituents including anthraquinone derivatives (naphthodianthrones), flavonoids, prenylated phloroglucinols, tannins and volatile oils is also included. Various types of preparations, ointments, creams and extracts prepared with and compounds isolated from this species have been found to possess a broad spectrum of biological and pharmacological effects such as antidepressant effects, wound-healing, antiviral and antimicrobial activity. The antibacterial activity of crude extracts can be related to the use of the herb as a wound healer in ancient times. The sole antibacterial principle isolated to date is a tetraketone, hyperforin, also thought to be responsible for the antidepressant activity of the herb. The available literature indicates that it has a higher antibacterial activity against Gram-positive than Gram-negative bacteria, and alcoholic extracts (methanolic/ethanolic) were shown to possess more pronounced activity than aqueous extracts. Based on the chemical and pharmacological characteristics of H. perforatum, we concluded that this species has beneficial therapeutic properties and has the potential for use as an effective adaptogenic herbal remedy.