Brine shrimp lethality and cytotoxicity assay of Araucaria bidwillii Hook in human carcinoma cell lines

Ahamed, KFH Nazeer,Kumar, V,Manikandan, L,Wahile, Atul M,Mukherjee, Kakali,Saha, BP,Mukherjee, Pulok K Kyung Hee Oriental Medicine Research Center 2005 Oriental pharmacy and experimental medicine Vol.5 No.1

The leaf extracts of Araucaria bidwillii Hook. (Araucariaceae) were evaluated for their cytotoxic effect in various human cancer cell lines. Preliminary investigation by brine shrimp lethality assay indicated that $LC_{50}$ value of various successive extracts were found to be less than $1000\;{\mu}g/ml$, where the ethyl acetate extract showed maximum activity of less than $100\;{\mu}g/ml$. Further cytotoxic evaluation of various leaf extracts of Araucaria bidwilli Hook was carried out in four different human cancer cell lines-acute myeloblastic leukemia (HL-60), chronic myelogenic leukemia (K-562), breast adenocarcinoma (MCF-7) and cervical epithelial carcinoma (HeLa). Cytotoxicity was assessed by trypan blue dye exclusion method and 3-(4,5-dimethyl thiazole-2yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay. From the present investigation it was found that the ethyl acetate and methanol extract of Araucaria bidwilli Hook was found to be more effective in leukemic cell lines and was less effective in MCF-7 and HeLa. The $IC_{50}$ value of the ethyl acetate extract in leukemic cell lines was found to be $28.18\;and\;34.64\;{\mu}g/ml$ and methanol extract was found to be $33.11\;&\;39.81\;{\mu}g/ml$. It can be concluded that various extract from the leaves of Araucaria bidwillii Hook. posses cytotoxic activity tested in brine shrimps and various human carcinoma cell lines.

Antioxidant potential of aerial part of Asclepias curassavica. Linn (Family-Asclepiadaceae)

Raja, S,Ahamed, KFH Nazeer,Kumar, V,Mukherjee, Kakali,Bandyopadhyay, Arun,Mukherjee, Pulok K Kyung Hee Oriental Medicine Research Center 2005 Oriental pharmacy and experimental medicine Vol.5 No.2

Asclepias curassavica. Linn, an erect, simple (or) much branched perennial herb with a somewhat woody base, belonging to the family Asclepiadaceae. It has been reported to have multiple pharmacological effect of anti-inflammatory, cardiotonic, anticancer, anthelmintic and to treat piles and gonorrhoea. It is to be expected that several activities might be related to a possible antioxidant action from this plant. The hydro alcoholic extract of Asclepias curassavica was tested in vitro for its antioxidant activities, such as DPPH radical, nitric oxide radical, superoxide anion radical, lipid peroxidation assay, hydroxyl radical, reducing power, and total phenol content. The extract exhibited scavenging potential with $IC_{50}$ value of $8.7\;{\mu}g/ml,\;198.4\;{\mu}g/ml\;and\;21.7\;{\mu}g/ml$ for DPPH, nitric oxide and superoxide anion radicals. The values were found to higher than those of Vitamin-C, rutin, and curcumin, as standards. The extract showed 50% protection at the dose of $134.2\;{\mu}g/ml\;and\;41.4\;{\mu}g/ml$ in lipid peroxidation as well as deoxyribose degradation, those values more to that of standard, vitamin E $(IC50\;values,\;119.2\;{\mu}g/ml\;and\;32.5\;{\mu}g/ml,\;respectively)$. The reducing power of the extract depends on the concentration and amount of extract. Since a significant amount of polyphenol could be detected by the equivalent to $0.0495\;{\mu}g$ of pyrocatechol from 1 mg of extract. It can be concluded that hydro alcoholic extract of aerial part of Asclepias curassavica could be considered as potent antioxidant, which makes it suitable for the prevention of human disease.

Reversible histoarchitecture study of testis and cauda epididymis andchanges in cauda epididymal epithelial cell types on treatment with benzeneextract of Ocimum sanctum leaves in albino rats

Mukhtar Ahmed,R Nazeer Ahamed,RH Aladakatti,Deepthi KR 경희대학교 융합한의과학연구소 2008 Oriental Pharmacy and Experimental Medicine Vol.8 No.2

In the present study, an attempt has been made to assess whether the effect of benzene extract of Ocimum sanctum leaves on the ultrastructural changes in the epithelial cells of the cauda epididymis, its subsequent recovery in the seminiferous epithelium and fertility of male albino rats. Wistar strain male albino rats were orally administered benzene extract of 250 mg/kg body weight of O. sanctum leaves followed by subsequent recovery maintaining suitable controls for 48 days. Results indicate decrease in the weights of testis, epididymis and seminal vesicles. Other accessory organs were not affected. Total count, cell and nuclei diameters of germ cells and Leydig cells were reduced. Cauda epididymis exhibited significant reduction in epithelial height and nuclei diameter of epithelial cells. Cells showed vacuolization with exhibit of signs of degeneration. Ultra study revealed that, in general, the cauda epididymis was affected and in particular, the principal, clear and basal cells were highly disturbed. Further, there was decrease in the size of lipid droplets, mitochondria, Golgi complex, endoplasmic reticulum and accumulation of lysosomal bodies. Fertility performance test showed no implantation in female rats mated with O. sanctum treated rats. Moreover, their recovery after withdrawal of treatment was observed suggesting that the effect of the treatment is transient and reversible. A recovery period resulted in normal spermatogenesis and fertility, suggesting reversible antispermatogenic and antifertility effects of the plant. In the present study, an attempt has been made to assess whether the effect of benzene extract of Ocimum sanctum leaves on the ultrastructural changes in the epithelial cells of the cauda epididymis, its subsequent recovery in the seminiferous epithelium and fertility of male albino rats. Wistar strain male albino rats were orally administered benzene extract of 250 mg/kg body weight of O. sanctum leaves followed by subsequent recovery maintaining suitable controls for 48 days. Results indicate decrease in the weights of testis, epididymis and seminal vesicles. Other accessory organs were not affected. Total count, cell and nuclei diameters of germ cells and Leydig cells were reduced. Cauda epididymis exhibited significant reduction in epithelial height and nuclei diameter of epithelial cells. Cells showed vacuolization with exhibit of signs of degeneration. Ultra study revealed that, in general, the cauda epididymis was affected and in particular, the principal, clear and basal cells were highly disturbed. Further, there was decrease in the size of lipid droplets, mitochondria, Golgi complex, endoplasmic reticulum and accumulation of lysosomal bodies. Fertility performance test showed no implantation in female rats mated with O. sanctum treated rats. Moreover, their recovery after withdrawal of treatment was observed suggesting that the effect of the treatment is transient and reversible. A recovery period resulted in normal spermatogenesis and fertility, suggesting reversible antispermatogenic and antifertility effects of the plant.

Dose dependent effect of benzene extract of Ocimum sanctum leaves on cauda epididymal spermatozoa of albino rats

Mukhtar Ahmed,R Nazeer Ahamed,RH Aladakatti,Ghodesawar MG 경희대학교 융합한의과학연구소 2009 Oriental Pharmacy and Experimental Medicine Vol.9 No.4

An attempt has been made to assess whether the dose dependent effect of benzene extract of Ocimum sanctum leaves on the morphological changes in the cauda epididymal spermatozoa and sperm parameters in male albino rats. Scanning Electron Microscope observations illustrate the disturbance in plasma membrane as well as acrosomal membrane. Most of the sperms appear morphologically abnormal in the mid region of the tail; there is formation of balloon like cytoplasmic droplet. Sperm parametric study exhibits decrease in the total sperm count, sperm motility, forward velocity and increase in the percentage of abnormal sperms in dose dependent manner on treatment benzene extract of Ocimum sanctum leaves. The results suggest that the effects may have resulted from a general disturbance in the proteins and alteration in cauda epididymal milieu probably due to androgen deficiency consequent upon antiandrogenic property of Ocimum sanctum leaves. An attempt has been made to assess whether the dose dependent effect of benzene extract of Ocimum sanctum leaves on the morphological changes in the cauda epididymal spermatozoa and sperm parameters in male albino rats. Scanning Electron Microscope observations illustrate the disturbance in plasma membrane as well as acrosomal membrane. Most of the sperms appear morphologically abnormal in the mid region of the tail; there is formation of balloon like cytoplasmic droplet. Sperm parametric study exhibits decrease in the total sperm count, sperm motility, forward velocity and increase in the percentage of abnormal sperms in dose dependent manner on treatment benzene extract of Ocimum sanctum leaves. The results suggest that the effects may have resulted from a general disturbance in the proteins and alteration in cauda epididymal milieu probably due to androgen deficiency consequent upon antiandrogenic property of Ocimum sanctum leaves.

Hypericum source of natural antimicrobials

Mukherjee, Pulok K.,Wahile, Atul,Ahamed, KFH Nazeer,Rajan, S. Kyung Hee Oriental Medicine Research Center 2003 Oriental pharmacy and experimental medicine Vol.3 No.3

Plants of the genus Hypericum (Family - Hypericaceae) are herbs, shrubs or small trees and are distributed chiefly in the temperate regions of the world. About 400 different species of Hypericum are available throughout the globe and 20 species occur in India, including a few cultivated in gardens. Almost all plants of the genus Hypericum are widely used in folk medicine. Several potent phytoconstituents from different Hypericum species have led to the isolation of antibacterial, antifungal and cytotoxic compounds. With the development of resistance and cross resistance with different microorganisms and the evolution of so many deadly diseases the screening and evaluation of the phytoconstituents so much so the development of varied phytoconstituents for the drug development for these deadly diseases is utmost essential in every aspects. The present review on the antimicrobial use of different Hypericum reports the findings from and extensive literature search on the Hypericum species around the globe that have been assessed for antimicrobial and antiviral activity. An attempt has been made through this review to summarize the information in this aspect in order to highlight the promising species of this genus which are worthy for further investigation as leads for drug development. Over 31 different Hypericum species have been reported to possess such activities with their varied number of phytoconstituents. Sixteen different constituents of six different classes of phytoconstituents have been reported to be present in different varieties of Hypericum, which may be considered responsible for this activity.