[10] Experimental animals Wistar albino mice of either sex weighing 35�C40 g, bred in Central example Animal House facility of the institute, were used for this study. The animals were housed under standard laboratory conditions, maintained on natural light and dark cycle, and had free access to food and water. They were acclimatized to laboratory conditions before the experiment. Each animal was used once in every experiment, and all experiments were carried out in daylight. Acute toxicity study Acute toxicity study was carried out according to the Organization for Economic Co-Operation and Development (OECD) Guidelines No. 423. Three animals were used for each step. The dose level to be used as the starting dose was selected from one of four fixed levels, i.e., 5, 50, 300, and 2000 mg/kg body weight p.

o. As per the OECD recommendations, the starting dose level should be that which is most likely to produce mortality in some of the dosed animals; and when there is no information available on a substance to be tested in this regard; for animal welfare reasons, recommended starting dose is 300 mg/kg body weight. Hence, we have used 300 mg/kg body weight as starting dose. Neuropharmacological study Six animals were used in each group for each experiment separately. Animals treated with 5% gum acacia suspension (0.1 ml p.o.) served as control. Diazepam (2 mg/kg i.p.) served as standards, and animals in test group were treated with EEAA (400 mg/kg i.p.), respectively. Each animal was treated with respective drug 30 min before experiment.

Tests performed were as GSK-3 follows: Rota-rod performance Four animals at a time were placed on rod rotating at 20�C25 rpm speed. Only the mice that demonstrated their ability to remain on the revolving rod (20�C25 rpm) for 5 min after training sessions during pretest screening were selected for studies. The fall off time was recorded in all the groups before and 30 min after drug administration. Decrease in fall off time is suggestive of depression of the central nervous system (CNS).[11] Actophotometer test The animal locomotor behavior was monitored using actophotometer. Animals were placed in actophotometer individually, and basal activity score was recorded over the period of 5 min. Each animal was treated with respective drug, and activity score was recorded after 30 min and 1 h. Decreased activity score was taken as index of CNS depression.[12�C14] Open field test Open field apparatus was designed as described by Gray and Lalji (1971) with few modifications. Dimensions were 50 �� 50 �� 40 cm made up of plywood open from top and bottom kept on white table top; surface was divided into 25 equal squares, i.e., 9 central and 16 peripheral.