Table 1: Antifungal Activity of Some Plant Extracts against 25 Isolates of Candida Species Evaluation of Cns Depressant Activity of Different Plant Parts of Nyctanthes Arbortristis Linn

Candida is a difÞ cult organism to establish standards of therapeutic activity. The problem is compounded for plant extracts in which variation may be expected between samples depending on genotype, area of cultivation, time of harvesting, processing methods, dilution etc. The extracts in our study are crude and that may be the reason for the antifungal activity of these extracts up to 100 mg/ml for the S. jambolanum, C. siamea and C. scalpelliformis. Probably, a more refined preparation would have antifungal activity at a lower concentration. Further experimental and clinical trials should be carried out and if these antifungals are effective they could be used as therapeutic agents. (mg/ml) (mg/ml) (mg/ml) (mg/ml) (mg/ml)a indicates number of isolates;-indicates no activity; 1+ indicates zone of inhibition in average of 7 to 10 mm; 2+ indicates zone of inhibition in average of 11 to 14 mm; 3+ indicates zone of inhibition in average of 15 to 18 mm; 4+ indicates zone of inhibition in average of 19 to 22 mm. Das, et al.: CNS depressant activity of Nyctanthes arbortristis Linn.

Nyctanthes arbortristis Linn.(Family Oleaceae), commonly known as Harsingar or Night Jasmine, is a common wild hardy large shrub or small tree 1,2 .
Nyctanthes arbortristis L. is used by the rural people of Orissa in India to cure various ailments along with its use in Ayurveda, Sidha and Unani systems of medicines.It's claimed traditional uses have been proved on scientiÞ c basis using in vitro and in vivo experiments [3][4][5][6][7] .It has been established that its leaves possess hypnotic and tranquillizing activities 3,4 and its ß owers possess sedative activity 5 .The present study is aimed at the CNS depressant activity of the ethanol extracts of different parts (ß owers, barks, seeds and leaves) of Nyctanthes arbortristis L. along with an attempt explore the responsible mechanism.
The ß owers, barks, seeds and leaves of Nyctanthes arbortristis L. were collected from the garden of BIT, Mesra, Ranchi and forests of Orissa.The herbarium of the plant (CNH/I-I (20)/2005-Tech-II/254) was authenticated as Nyctanthes arbortristis L. from Botanical Survey of India, Kolkata.After drying properly, the leaves, barks and seeds were powdered coarsely and then were extracted successively with petroleum ether, chloroform and ethanol (90%) 6,7 , whereas its fresh flowers with ethanol (50%) 7 .The ethanol extracts were evaporated to dryness, having yield values 14%, 12.5%, 26.5% and 13% w/w, respectively.The water-soluble portions of the extracts were subjected to the pharmacological screening.
Adult male swiss mice weighing between 20-30 g, obtained from the animal house of BIT, Mesra, Ranchi, were used for this investigation.The Institutional Animals Ethics Committee (Registration No. 62/02/ac/CPCSEA) approved the experiments.Up and down or staircase method was followed for the estimation of acute toxicity of the water-soluble portion of the ethanol extracts of different parts of Nyctanthes arbortristis L. The dose was increased from 400 mg/kg to 2.0 g/kg, through intraperitoneal route of administration 8,9 .
The gross behavioral study was performed 30 min after the administration of the extracts to get maximum information about the effect of the extracts on the central nervous system of mice 9 .The control group of animals was only treated with pentobarbital sodium (45 mg/kg).Each extracts and the reference compound were injected 30 min before pentobarbital sodium administration.The time taken for the loss of righting reß ex was noted in all cases.The onset of sleep was recorded by noting the time of loss of righting reflex of mice and duration of sleep by noting time difference between loss of righting reß ex and recovery time 10 .
The monoamine neurotransmitters in brain were estimated following the method described by Shellenderger et al. 11 The animals were sacriÞ ced half an hour after the administration of the extracts by cervical dislocation.After chilling the head of mice in chilling mixture of ice and CaCl 2 , their brains were taken out and weighed 11,12 .
For the estimation of dopamine, brains were homogenized with dry n-butanol at 0 o .Clear supernatant solutions were extracted with 0.1 M phosphate buffer.To the phosphate buffer extract 4% EDTA, Iodine solution, alkaline sulÞ te and 5N acetic acid were added and then heated.After cooling, the intensities of fluorescence were determined by the help of photoß uorometer 11,12 .
For the estimation of serotonin, brains were homogenized with 0.1N HCl at 0 o and then centrifuged.To the clear supernatant solutions, 10% zinc sulfate and 1N NaOH were added and then centrifuged.To a quartz cuvette containing 2N HCl the supernatant fluid was added and its intensity of the ß uorescence was determined by the photofluorometer 11,12 .The results were expressed in mean±standard error of mean.The results were subjected to statistical analysis, using ANOVA to determine the signiÞ cance of the tested activity and p<0.001 were considered to be signiÞ cant 10 .
Toxicity studies revealed that the maximum tolerable dose for the water-soluble portion of the ethanol extracts was more than 2.0 g/kg body weights.
From which, doses of 200, 400 and 600 mg/kg were selected for the evaluation of CNS depressant activity of the extracts.The gross behavioral study on mice of the extracts showed significant CNS depressant activity and some muscle relaxant activity.The leaves, ß owers and seeds showed signiÞ cant (p<0.001) and dose-related prolongation of the onset and duration of sleep, which are well comparable to that of the standard drug chlorpromazine (Table 1).It has been observed that the leaves possess highest CNS depressant activity.
The changes in dopamine and 5-hydroxytryptamine (5-HT) levels, expressed as percentage increase or decrease from their respective control values, are presented in Table 2.The ethanol extracts of the leaves, seeds and flowers of Nyctanthes arbortristis L. were found to cause decrease in dopamine and increase in 5-HT level significantly in a dose-dependent manner.Its barks were found to cause these changes only at its higher dose of 600 mg/kg.Decrease in dopamine level may lead to the development of catalepsy and depression in the brain 13 .This can be well correlated with the antipsychotic activity of the extracts.
Many antipsychotic agents also have some affinity for 5-HT receptors and drugs, which increase brain 5-HT and usually increase sleep 14 .This is evident in case of the extracts treatment where the 5-HT level is increased.This can be well correlated with the potentiation of pentobarbitone-induced sleeping time in mice by the extracts, which could be accounted to

TABLE 2: EFFECT OF THE WATER-SOLUBLE FRACTION OF THE ETHANOL EXTRACTS OF NYCTANTHES ARBORTRISTIS ON MONOAMINE NEUROTRANSMITTERS IN THE BRAIN OF MICE
the increase in the concentration of 5-HT in the brain.
The overall study conÞ rms that the CNS depressant action and the potentiation of pentobarbitone-induced sleeping time by the ethanol extracts of flowers, seeds, leaves and barks (600 mg/kg) of Nyctanthes arbortristis L. might be due to the decrease in dopamine and increase in the 5-hydroxytryptamine level in brain.

TABLE 1 : ANTIFUNGAL ACTIVITY OF SOME PLANT EXTRACTS AGAINST 25 ISOLATES OF CANDIDA SPECIES
indicates number of isolates; -indicates no activity; 1+ indicates zone of inhibition in average of 7 to 10 mm; 2+ indicates zone of inhibition in average of 11 to 14 mm; 3+ indicates zone of inhibition in average of 15 to 18 mm; 4+ indicates zone of inhibition in average of 19 to 22 mm. a

TABLE 1 : PROLONGATION OF PENTOBARBITAL- INDUCED SLEEPING TIME BY THE ETHANOL EXTRACTS OF DIFFERENT PARTS OF NYCTANTHES ARBORTRISTIS LINN.
Values are expressed in Mean±SEM, n = 6, *p<0.001.NAF, NAB, NAS and NAL represent the ethanol extracts of the ß owers, barks, seeds and leaves of Nyctanthes arbortristis respectively.