Hepatoprotective activity of the neem-based constituent azadirachtin-A in carbon tetrachloride intoxicated Wistar rats.

The aim of this study was to investigate the hepatoprotective role of azadirachtin-A in carbon tetrachloride (CCl4) induced hepatotoxicity in rats. The group allotment for the animals used in the hepatoprotective study included a vehicle treatment group, CCl4 (1 mL · (kg body mass)(-1)) treatment group, silymarin (100 µg · (kg body mass)(-1) · day(-1)) + CCl4 treatment group, and groups treated with different doses of azadirachtin-A (100 or 200 µg · (kg body mass)(-1) · day(-1)) + CCl4. On the 9th day, blood was obtained for measuring the biochemical parameters, and liver tissue was obtained for pathological examination. The acute toxicity test with azadirachtin-A (500, 1000, or 2000 µg · (kg body mass)(-1)) indicated no mortality after 14 days of treatment; further, there was no change in behavior, food consumption, or organ mass. However with the higher dose, some hematological parameters showed changes. Hepatoprotective studies revealed that the CCl4 treatment group exhibited a decrease in total protein and albumin levels, whereas a significant increase in BUN, AST, ALT, and ALP levels were noticed compared with the vehicle-treated control, indicating that there was liver damage caused by CCl4. Histology and ultrastructure study confirmed that pretreatment with azadirachtin-A dose-dependently reduced hepatocellular necrosis and, therefore, protected the liver against toxicity caused by CCl4. The results from this study indicate that pretreatment with azadirachtin-A at the higher dose levels, moderately restores the rat liver to normal. This study confirms that azadirachtin-A possesses greater hepatoprotective action; however, the effective concentration needs to be determined.

Effect of phenol on oxygen consumption and bioaccumulation in different tissues of freshwater fish Cyprinus carpio.

The influence of phenol exposure history on the rates of phenol accumulation, elimination, and oxygen consumption was examined in the fresh water fish species Cyprinus carpio. Bioaccumulation and elimination of phenol from the kidney and biochemical parameters of liver gill and muscle were determined at intervals of 10, 20, and 30 days. A significant increase in phenol concentration was noted in all treated groups. The acute toxicity of phenol was 30.0 mg/L. Changes were dose and duration dependant when fish were exposed for 30 days to two sublethal concentrations of phenol (1.5 and 3.0 mg/L). Recovery post exposure was observed after transfer to normal tap water for 30 days. Elimination of phenol was noted, although the concentration remained significantly higher than the control. The longer the phenol exposure time, the greater was the percentage reduction of organic matter of the fish. Total protein, total carbohydrate, and total lipids in the tissues of liver, gill, and muscle decreased significantly. On the other hand, these parameters were higher at the end of the experiment than at the start (ANOVA P < or = .01).

Effect of benzene extract of Ocimum sanctum leaves on cauda epididymis of albino rats.

In the present study, an attempt has been made to assess the effect of benzene extract of Ocimum sanctum leaves on ultrastructural changes in epithelial cells of the cauda epididymis and fertility of male albino rats. Wistar strain male albino rats were orally administered 250 mg/kg body weight of O. sanctum leaves followed by maintaining suitable controls for 48 days. The results indicate, in cauda epididymis, a significant reduction in epithelial height and nuclei diameter of epithelial cells. Cells showed vacuolization exhibiting of signs of degeneration. An ultra study revealed that, in general, the cauda epididymis was affected and in particular, the principal, clear and basal cells were highly disturbed. Further, the size of lipid droplets, mitochondria, Golgi complex, endoplasmic reticulum decreased, with an accumulation of lysosomal bodies. The fertility performance test showed no implantation in female rats mated with O. sanctum-treated male rats. The results suggest a probable dwindling of an androgen status/direct effect on the above observations.

Tyrosine phosphorylated proteins in mammalian spermatozoa: molecular and functional aspects.

During mammalian fertilization, spermatozoa must undergo capacitation and the acrosome reaction. These processes of sperm function are critically associated with various molecular events and one such process is protein tyrosine phosphorylation (PYP). This event is downstream of increases in intracellular Ca2+ and activities of HCO3- activated adenylate cyclase, cAMP-dependent-protein kinase-A and reactive oxygen species. Though, PYP is known to be mediated by tyrosine kinases and phosphatases, only a few of them have been identified and characterized in spermatozoa. Since most identified tyrosine kinases are soluble proteins from somatic cells, it is believed that distinct mechanisms could exist in spermatozoa for PYP. Such sperm-specific protein tyrosine kinases/ phosphatases still remain to be thoroughly characterized in most species, including hamsters. Nevertheless, a few tyrosine phosphorylated sperm proteins have been identified in hamsters and in other mammals as well. There is very limited information available on our understanding of the molecular and ultrastructural localization, as well as the characteristics of tyrosine phosphorylated proteins. Functionally, how sperm motility is regulated by PYP is also poorly understood. Knowledge of tyrosine phoshorylated proteins and how they regulate sperm function is of immense significance in our understanding of male (in)fertility and clinical management of fertility; especially, in the light of studies that implicate the hypo-tyrosine phosphorylated state of sperm proteins with asthenozoospermic condition in humans. This article provides a comprehensive review on PYP and its regulation by kinases and phosphatases.

Toxicity, bioaccumulation and metabolism of phenol in the freshwater fish Oreochromis mossambicus.

The acute toxicity of phenol was determined to be 35.0 mg/l in Oreochromis mossambicus. The fish were exposed to two sublethal concentrations of phenol (2.3 and 3.5 mg/l) for 30 days. The effects of exposure were studied on the bioaccumulation and elimination of phenol from the kidney and biochemical parameters of liver, gill, and muscle at intervals of 10, 20, and 30 days. A statistically significant increase in phenol concentration was noted in tissues from all treated fish groups. Bioaccumulation and biochemical changes were dose and duration dependant. Recovery in fish after post exposure was observed after transferring these fish to normal tap water for 30 days. Elimination of phenol was noted, although the concentration of phenol remained significantly higher than the control after 30 days of the experiment. Total protein, total carbohydrate, and total lipids in the tissues of liver, gill, and muscle of fish decreased greatly. The longer the exposure time, the greater was the percentage reduction of organic matter of the fish exposed to the sublethal concentration of phenol.

Effect of phenol on the metabolism and biochemical composition of the freshwater fish Cyprinus carpio.

The effect of phenol on the fish Cyprinus carpio and the metabolic changes in oxygen consumption and total carbohydrate, total protein and total lipids in the tissues of liver, gills, muscle, brain, and intestine were investigated for sublethal exposure. The oxygen consumption decreased significantly in both toxicants exposed to acute toxicity. Following sublethal exposure to phenol total protein, total carbohydrate, and total lipids in the tissues of liver, gills, muscle, brain and intestine of the fish decreased greatly. The longer the exposure time, the greater was the percentage reduction of organic matter of the fish exposed to sublethal concentration of phenol. Lipid content was not reduced much in exposure when compared with the control.

Effect of graded doses of Momordica charantia seed extract on rat sperm: scanning electron microscope study.

Morphological changes in sperm of albino rats observed under scanning electron microscopy illustrate the disturbance in the plasma membrane as well as in the acrosomal membrane on treatment with effect of graded doses of alcohol seed extract from Momordica charantia. Considerable changes in the shape and size of the sperm head were observed, with the middle region of the sperm head being slightly constricted dorsoventrally. Most sperm appeared morphologically abnormal in the mid-region of the tail, with formation of a balloon-like cytoplasmic droplet. The results of this study suggest that such effects may have resulted from a general disturbance in proteins and an alteration in the cauda epididymal milieu, probably due to androgen deficiency consequent to the anti-androgenic property of Momordica charantia seeds.

Changes in Sertoli cells of albino rats induced by Azadirachta indica A. Juss leaves.

Azadirachta indica leaf powder, 0.05 mg testosterone, or combined leaf powder + testosterone was administered for 48 days to different groups of 3-month-old albino rats. Twenty-four hours after the final dose, the treated animals received mild ether anesthesia and the testes were dissected out and processed for light and electron microscope studies. Animals treated with A. indica leaf powder showed damaged tubules and exhibited an abundance of vacuoles, including inter-cellular spaces and intra-epithelial vacuoles of varying size in the cytoplasm of Sertoli cells. Upon treatment with A. indica leaf powder, bridges between Sertoli cells-Sertoli cells and Sertoli cell-germ cells were disturbed, coupled with changes in the Sertoli cells and cytoplasm along with its organelles. We suggest that the anti-androgenic property of A. indica leaves probably affects Sertoli cells, followed by the degeneration of germ cells, resulting in exfoliation.

Ultrastructural changes in cauda epididymidal epithelial cell types of Azadirachta indica leaf treated rats.

To assess if cauda epididymis is a target for the effect of A. indica leaves, Wistar strain male albino rats were administered (po) A. indica leaves (100 mg/rat/day for 24 days). Transmission electron microscopic analysis revealed that in the cauda epididymal epithelium the nuclei of principal cells were enlarged and the number of coated micropinocytotic vesicles of the apical cytoplasm decreased. Microvilli were missing and mitochondrial cristae and Golgi complex were highly disrupted. The cytoplasm was abounding with lysosomal bodies. The clear cells increased in perimeter and their nuclei increased in size and contained lesser chromatin. The nuclear membrane bulged out. The cytoplasm was vacuolized. Further, there was decrease in size of the lipid droplets, mitochondria, Golgi complex, endoplasmic reticulum and there was accumulation of lysosomal bodies. The changes in the principal and clear cells appear to be due to the effect of the hypoandrogen status caused by treatment with A. indica leaves and a direct action on the epididymal epithelium.

Azadirachta indica adversely affects sperm parameters and fructose levels in vas deferens fluid of albino rats.

Azadirachta indica treatment for 24 days in albino rats resulted in a decrease in the total sperm count, sperm motility, and forward velocity in vas deferens fluid. The percentage of abnormal sperm increased and the fructose content decreased. As diminished levels of fructose parallel androgen deficiency, we conclude that reduced androgen levels resulting from the anti-androgenic property of A. indica leaves probably influences the physiological maturation of sperm.

Reversible anti-fertility effect of benzene extract of Ocimum sanctum leaves on sperm parameters and fructose content in rats.

Treatment of albino rats with a benzene extract of Ocimum sanctum leaves (250 mg/kg body weight) for 48 d decreased total sperm count, sperm motility, and forward velocity. The percentage of abnormal sperm increased in caudal epididymal fluid, and the fructose content decreased in the caudal plasma of the epididymis and the seminal vesicles. The results suggest that such effects are due to androgen deprivation, caused by the anti-androgenic property of O. sanctum leaves. The effect was reversible because all parameters returned to normal 2 wk after the withdrawal of treatment.

Ultrastructural changes induced by leaves of Azadirachta indica (Neem) in the testis of albino rats.

The present work was designed to study the effect of Azadirachta indica (Neem) powder on rat testis using the electron microscope. Male albino rats received 100 mg each A. indica leaf powder orally (by gavage). On alternate days, a second group of rats received 0.125 mg testosterone dipropionate intramuscularly. A third group received both A. indica leaf powder by gavage and testosterone dipropionate intramuscularly. Suitable controls were maintained. After autopsy, ultrastructural analysis of the testis revealed that animals treated with testosterone dipropionate showed well-developed Sertoli cells and germ cells with well-developed cytoplasmic organelles. By contrast, in A. indica-treated rats, intracellular spaces and vacuolization were observed in Sertoli cells; whereas in Leydig cells, cytoplasmic inclusions appeared diminished, and the configuration of granular endoplasmic reticulum appeared as a single unbranched tubule. In late spermatids, defects were observed in the mitochondrial sheath. The ultrastructural changes seen in the A. indica-treated group provide a clue that A. indica leaves might affect spermatogenesis through antispermatogenic and antiandrogenic properties.

Sperm parameters changes induced by Azadirachta indica in albino rats.

Azadirachta indica treatment for 48 days In albino rats resulted in a decrease in the total sperm count, sperm motility, and forward velocity. The percentage of abnormal sperm increased and the fructose content of caudal semen of the epididymis decreased. The observations suggest that these effects are probably due to an androgen deficiency, caused by the anti-androgenic property of the leaves of A. indica, thereby affecting the physiological maturation of sperm.

Effect of Azadirachta indica leaves on rat spermatozoa.

Histochemical studies and SEM observations on the morphological changes in the head of the spermatozoa in general, and the acrosome in particular, in A. indica treated rats are reported. In the treated rats change in the shape and size of the sperm head, with a dorso-ventral constriction of the middle region of the sperm head i.e., between the anterior and posterior regions was observed. It was rather difficult to differentiate the outer acrosomal and outer plasma membranes. A decrease in the perforatorium or sub-acrosomal material, post nuclear cap and the nuclear material near the basal plate at the base of the sperm head were also observed. The results suggest that the effects are probably due to androgen deficiency and a general disturbance in carbohydrates or polysaccharides located in the sperm head, caused by the antiandrogenic property of the leaves of A. indica.