M. A. Akbarsha

A herbal medicine for the treatment of lung cancer

Lung cancer is the leading cause of cancer-related deaths throughout the world. Extracts of medicinal plants are believed to contain different chemopreventive or chemotherapeutic compounds. In this study, we determined the anti-cancer property of one of the traditional Indian medicine Rasagenthi Lehyam (RL) for the treatment of lung cancer. Two lung cancer cell lines (A-549 and H-460) and one normal bronchial epithelial (BEAS-2B) cell line were used to test the chemotherapeutic effect of RL. Out of five fractions of RL, chloroform fraction of RL (cRL) demonstrated a significant inhibition of cell proliferation and induction of apoptosis in A-549 and H-460 cells but not in normal BEAS-2B cells. The cRL fraction up-regulated the pro-apoptotic genes p53 and Bax and induced caspase-3 activation, and down-regulated the pro-survival gene Bcl-2 in both the lung cancer cell lines. Also, nuclear export of p53 was seen in cRL-treated lung cancer cells. In addition, cRL induced G2/M arrest of cell cycle and enhanced the radio-sensitivity of both the lung cancer cell lines. This study suggests that cRL may prove to be a potent anti-cancer agent that may be used for the treatment of lung cancer. However, further studies are required to bring cRL into the mainstream of medicine in the treatment of lung cancer. (Mol Cell Biochem xxx: 125–133, 2005)

Seasonal variation in spermatogenic and androgenic activities in a caecilian testis (Ichthyophis tricolor)

Caecilians are an order of amphibians with distribution confined to several of the tropical countries. They are subterranean animals and practice a form of internal fertilization. Several species are viviparous. Published reports on seasonal variation in male reproduction in caecilians are limited to a very few species. A study was undertaken to examine the seasonal variation of testicular activity with respect to spermatogenesis and steroidogenesis in a caecilian, Ichthyophis tricolor , fairly abundant in the Western Ghats of Kerala, India, using light and transmission electron microscopy, terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL) staining and biochemical determination of 3-hydroxy-Δ 5 -steroid dehydrogenase (Δ 5 -3β-HSDH) activity. Three phases in spermatogenic activity, active spermatogenesis (July–November), early regression (December–March) and spermatogenic quiescence (April–June) are identified. Apoptosis seems to be the mechanism of germ cell death during the regression phase. Leydig cells are interstitial and not peritubular. Activity of testicular Δ 5 -3β-HSDH follows the same pattern as spermatogenic activity, although histometric data on the interstitial tissue reveal higher areas during the regression and quiescent phases. Our findings will be useful in evolving strategies for conservation of caecilians with special reference to Kerala, India.

Stages in follicle cell/oocyte interface during vitellogenesis in caecilians Ichthyophis tricolor and Gegeneophis ramaswamii: a transmission electron-microscopic study

We describe the ultrastructural organization of the vitellogenic follicle stages in two caecilian species. Monthly samples of slices of ovary of Ichthyophis tricolor and Gegeneophis ramaswamii from the Western Ghats of India were subjected to transmission electron-microscopic analysis, with special attention to the follicle cell/oocyte interface. In order to maintain uniformity of the stages among the amphibians, all the stages in the caecilian follicles were assigned to stages I–VI, the vitellogenic and post-vitellogenic follicles being assigned to stages III–VI. Stage III commences with the appearance of precursors of vitelline envelope material in the perivitelline space. Stages IV and V have been assigned appropriate substages. During the transition of stage III to stage VI oocytes, a sequential change occurs in the manifestations of follicle cells, perivitelline space, vitelline envelope and oocyte cortex. The vitelline envelope becomes a tough coat through the tunnels of which the macrovilli pass to interdigitate between the microvilli. The oocyte surface forms pinocytic vesicles that develop into coated pits and, later, coated vesicles. Contributions of the oocyte cortex to the vitelline envelope and of the follicle cells to yolk material via synthesis within them are indicated. The follicle cell/oocyte interface of vitellogenic follicles of these two caecilians resembles that in anurans and urodeles, with certain features being unique to caecilians. Thus, this paper throws light on the possible relationships of caecilians to anurans and urodeles with special reference to ovarian follicles.

Assembly of ovarian follicles in the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii: light and transmission electron microscopic study

Though much is known about various aspects of reproductive biology of amphibia, there is little information on the cellular and mechanistic basis of assembly of ovarian follicles in this group. This is especially true of the caecilians. Therefore, taking advantage of the abundant distribution of caecilians in the Western Ghats of India, two species of caecilians, Ichthyophis tricolor and Gegeneophis ramaswamii, were subjected to light and transmission electron microscopic analysis to trace the sequential changes during the assembly of ovarian follicles. The paired ovaries of these caecilians are elongated sac-like structures each including numerous vitellogenic follicles. The follicles are connected by a connective tissue stroma. This stroma contains nests of oogonia, primary oocytes and pregranulosa cells as spatially separated nests. During assembly of follicles the oocytes increase in size and enter the meiotic prophase when the number of nucleoli in the nucleus increases. The mitochondrial cloud or Balbiani vitelline body, initially localized at one pole of the nucleus, disperses through out the cytoplasm subsequently. Synaptonemal complexes are prominent in the pachytene stage oocytes. The pregranulosa cells migrate through the connective tissue fibrils of the stroma and arrive at the vicinity of the meiotic prophase oocytes. On contacting the oocyte, the pregranulosa cells become cuboidal in shape, wrap the diplotene stage oocyte as a discontinuous layer and increase the content of cytoplasmic organelles and inclusions. The oocytes increase in size and are arrested in diplotene when the granulosa cells become flat and form a continuous layer. Soon a perivitelline space appears between the oolemma and granulosa cells, completing the process of assembly of follicles. Thus, the events in the establishment of follicles in the caecilian ovary are described.

Aspects of Male Reproductive Toxic Effects of Quassia amara L.: Histopathological and Ultrastructural Study in Mouse

Quassia amara is a Brazilian medicinal plant traditionally used as an herbal remedy for a variety of diseases. Quassin is the major secondary chemical compound in it, which is under investigation as an anti-malarial compound. There are preliminary studies implicating Q. amara in male reproductive toxicity. Hence, this elaborate study was undertaken to find the male reproductive toxic effects of Q. amara and quassin. Methanolic extract of Q. amara bark and quassin were administered to male mice for 35 days. The cauda epididymidal sperm, testis and epididymis were subjected to analysis adopting light and transmission electron microscopy. Sperm counts, motility and viability decreased. The incidence of sperm with abnormal morphologies increased. There was premature loss of germ cells from the seminiferous tubules, which correlated with Sertoli cell pathology. The Leydig cells were vacuolated, suggesting a hypo-androgen status. The epididymis indicated severe histopathological changes, including reflection of a mitogenic/tumorigenic effect of Q. amara . Thus, this study indicates that Q. amara , when applied as a therapeutic, could be severely toxic to male reproductive mechanisms and the principal mechanism of action appears to be through Leydig cells. Quassin, the major secondary metabolite responsible for the therapeutic potential of Q. amara , is also the one causative of the male reproductive toxic effects. Thus, this study substantiates the claim that Q. amara is toxic to male reproductive system.