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Analgesic and antidiarrhoeal activities of Treama orientalis Linn. in mice

June 2008
Oriental Pharmacy and Experimental Medicine 8(2):187-191

DOI:10.3742/OPEM.2008.8.2.187

Authors:

Sarder N. Uddin

MSU, USA

Khan Mohammad Ahsan Uddin

Aalto University

Firoj Ahmed

University of Dhaka

Trema orientalis Linn. is commonly grown in many parts of Bangladesh. Its leaves have been used for analgesic and anti-diarrhoeal activity in traditional medicine. This study evaluates the potential analgesic and anti-diarrhoeal activity of methanol and aqueous extracts of leaves in experimental acetic acid induced writhing and castor oil induced diarrhoea in mice. The aqueous extract of leaves showed significant (P < 0.001) analgesic effect in acetic acid induced writhing in mice at a dose of 500 mg/kg body weight In castor oil induced antidiarrhoeal screening both extract increased latent period (P < 0.025) and decrease the number of stool (P < 0.025) at the dose of 500 mg/kg body weight comparable with that of the standard drug loperamide. The results provide a support for the use of this plant in traditional medicine and suggest its further investigation.

. Effect of Trema orientalis on acetic acid induced writhing in mice (n = 4)

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. Effect of T. orientalis on castor oil induced diarrhea in mice (Latent Period)

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. Effect of Trema orientalis on castor oil induced diarrhea in mice (number of stools)

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2008 Kyung Hee University Press 1

Oriental Pharmacy and Experimental Medicine 2008 8(2), 00-00

www.opem.org

OPEM

Analgesic and antidiarrhoeal activities of Treama orientalis Linn. in mice

Sarder Nasir Uddin

, Khan Mohammad Ahsan Uddin

and Firoz Ahmed

Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh;

Pharmacy

Discipline, Khulna University, Khulna-9208, Bangladesh

Received for publication May 18, 2007; accepted October 16, 2008

SUMMARY

Trema orientalis Linn. is commonly grown in many parts of Bangladesh. Its leaves have been used for

analgesic and anti-diarrhoeal activity in traditional medicine. This study evaluates the potential

analgesic and anti-diarrhoeal activity of methanol and aqueous extracts of leaves in experimental acetic

acid induced writhing and castor oil induced diarrhoea in mice. The aqueous extract of leaves showed

significant (P < 0.001) analgesic effect in acetic acid induced writhing in mice at a dose of 500 mg/kg

body weight In castor oil induced antidiarrhoeal screening both extract increased latent period (P <

0.025) and decrease the number of stool (P < 0.025) at the dose of 500 mg/kg body weight comparable

with that of the standard drug loperamide. The results provide a support for the use of this plant in

traditional medicine and suggest its further investigation.

Key words: Analgesic; Antidiarrhoea; Trema orientalis Linn.; Extract

INTRODUCTION

During the past decade, traditional systems of

medicine have become increasingly important in

view of their safety. Current estimates suggest that,

in many developing countries, a large proportion

of the population relies heavily on traditional

practitioners and medicinal plants to meet primary

health care needs. Although modern medicine may

be available in these countries, herbal medicines

(phytomedicines) have often maintained popularity

for historical and cultural reasons. Concurrently,

many people in developed countries have begun to

turn to alternative or complementary therapies,

including medicinal herbs.

Bangladesh possesses rich floristic wealth and

diversified genetic resources of medicinal plants. It

has a widely ranging tropical and the agro climatic

conditions, which are conducive for introducing

and domesticating new and exotic plant varieties.

The use of the plants, plant extracts and pure

compounds isolated from natural sources provided

the foundation to modern pharmaceutical compounds.

T. orientalis (Bengali name- Jibon or Chikon) is a

tree and belongs to the Ulmaceae family. The plant

is distributed in almost all districts of Bangladesh

and is used in traditional medicine by the rural people

and possesses various interesting pharmacological

activities. The root of the plant is used in the treatment

of diarrhoea, asthma and passing of blood in urine;

the bark is used as poultice in muscular pain; the

roots, barks and leaves are used in epilepsy

(Kirtikar and Basu, 1980). In African folk medicine,

it is used in many diseases including dysentery,

*Correspondence: Sarder Nasir Uddin, Biotechnology

and Genetic Engineering Discipline, Khulna University,

Khulna-9208, Bangladesh. Fax: +88041731244; E-mail:

nasir713@yahoo.com

DOI 10.3742/OPEM.2008.8.2.001

2008 Oriental Pharmacy and Experimental Medicine 8(2), 00-00

2 Sarder Nasir Uddin et al.

hypertension, etc (Iwe, 1993). Fruit, leaves, bark,

stems, twigs and seeds are also used in traditional

medicine. The leaves are used to treat coughs and

sore throats and the bark is used to make cough

syrups. Other reported uses include remedies for

bronchitis, gonorrhoea, malaria, yellow fever, toothaches,

and intestinal worms (Rulangaranga, 1991).

As part of our continuing efforts to study the

chemical and pharmacological aspects of the

medicinal plants of Bangladesh, T. orientalis was

investigated Successive extraction of dried powdered

Leaves of T. orientalis (Family-Ulmaceae) methanol

and aqueous extracts and the effects of both extracts

were investigated analgesic and antidiarrhoeal

activities. Since there is a vast resource of Trema

Orientalis in Bangladesh, the present study will

provide some valuable information about the

pharmacological properties of this plant.

MATERIALS AND METHODS

Plant materials

Fresh leaves of T. orientalis were collected from

Khulna University Campus in Bangladesh. The

plant was identified by the experts of Bangladesh

National Herbarium, Mirpur, Dhaka (Accession

no. 31,285) and a voucher specimen was also

deposited there. The fresh leaves were cleaned,

dried and pulverized. About 400 g of powdered

material was taken in a clean, flat bottomed glass

container (4 l) and soaked in 1,300 ml of 80% of

methanol. The container was sealed and kept for a

period of 10 days with occasional shaking and

stirring. Then it was filtered and concentrated by

evaporation.

Experimental animal

Young Swiss-albino mice aged 4 - 5 weeks, average

weight 20 - 25 g were used for the experiment. The

mice were purchased from the animal Research

Branch of the International Centre for Diarrhoeal

Disease and Research, Bangladesh (ICDDR, B). They

were kept in standard environmental condition for

one week for adaptation after their purchase and

fed ICDDR, B formulated rodent feed and water.

Analgesic activity test

Analgesic activity of both methanol and aqueous

extract was tested by acetic acid induced writhing

model in mice (Whittle, 1964). Eighty four mice of

either sex were grouped in six (n = 14 per group).

Each group received a particular treatment i.e.

group-I for control, group-II for positive control,

test group-I for aqueous extract at the dose of 250

mg/kg, test group-II for aqueous extract at the

dose of 500 mg/kg, test group-III for methanol

extract at the dose of 250 mg/kg and test group-IV

for methanol extract at the dose of 500 mg/kg. 1%

tween-80 solution in deionized water at the doses

10 ml/kg per oral was used as control and

Diclofenac-Na was used at the doses of 25 mg/kg

body weights per oral as a standard positive

control. To prepare suspension of the test samples

at required doses, the required amount of extract

was triturated in unidirectional manner by the

addition of small amount of tween-80. After proper

mixing of extract and tweeen-80, the volume was

adjusted with deionized water. The test consists of

injecting 0.7% acetic acid solution intraperitoneally.

Control and test sample was given orally 30 min

prior to acetic acid injection. Each mouse of all

groups was observed carefully to count the number of

writhing that they had made in 15 minutes.

Incomplete writhing was taken as half-writhing, so

two half-writhing were taken as one full writhing.

Antidiarrhoeal acitvity test

Antidiarrhoeal activity was tested using the castor

oil induced diarrhea in mice (Chatterjee, 1993).

Experimental animals were randomly selected and

divided into four groups (n = 5 per group). Each

group received a particular treatment i.e. group-I

for control, group-II for positive control, test group-I

for aqueous extract and test group-II for methanol at

the dose of 500 mg/kg, Loperamid was used at the

doses of 4 mg/kg body weight as a standard.

Analgesic and antidiarrhoeal activities of Treama orientalis Linn. in mice 3

2008 Oriental Pharmacy and Experimental Medicine 8(2), 00-00

Control vehicle and the extract were administered

orally, 1h prior to the oral administration of castor

oil at dose of 0.5 ml per mice. Individual animals of

each group were placed in separate cages having

adsorbent paper beneath and examined for the

presence of diarrhea every hour in five hour study

after castor oil administered. Number of stool or

any fluid that stained the adsorbent paper was

counted at each successive hour during the

experiment. The latent period of each mouse also

counted.

Statistical analysis

Student’s t-test was used to determine a significant

difference between the control groups and

experimental groups for both the experiment.

RESULTS

The crude extract of aqueous and methanol extract

of leaves of T. orientalis linn. was used for analgesic

and antidiarrhoeal activity in mice at different

doses.

Analgesic activity of aqueous and methanol

extract of T. orientalis leaves was tested by acetic

acid induced writhing in mice. The methanol

extract showed 16.67% and 30.01% acetic acid

induced writhing inhibition in mice at the doses of

250 mg/kg and 500 mg/kg body weight respectively.

But the Aqueous extract showed 38.34% and

56.67% acetic acid induced writhing inhibition in

mice at the doses of 250 mg/kg and 500 mg/kg

body weight respectively. This was comparable to

diclofenac sodium where the inhibition was 48.34%

at the dose of 25 mg/kg body weight (Table 1).

In the study of antidiarrhoeal activity, aqueous

and methanol extract at the dose of 500 mg/kg

body weight was used as test group, 1% tween-80

at the dose of 10 mg/kg body weight was used as

control and loperamide at the dose 4 mg/kg body

weight was used as standard. From the study, it

showed that both aqueous and methanol extract

increased the latent period as compared to control

and standard. In Table 2, it was found that aqueous

extract and methanol extract increased the latent

period 1.456 h and 1.240 h respectively whereas

control and standard increased the latent period

1.096 h and 1.284 h respectively. From the same

doses we also showed that both the aqueous and

methanol extract decreased the number of stool as

Table 1. Effect of Trema orientalis on acetic acid induced writhing in mice (n = 4)

Animal group/Treatment

Number of Writhing

(% writhing)

Inhibition (%)

Group I- Control 1% Tween-80 solution in water, 10 ml/kg, p.o. 30 ± 1.07 (100) 00

Group II- Positive control-Diclofenac Sodium 25 mg/kg, p.o. 15.5 ± 0.64

(51.66) 48.34

Test group I- Aqueous extract 250 mg/kg, p.o. 18.5 ± 0.97

(61.66) 38.34

Test group II- Aqueous extract 500 mg/kg , p.o. 13 ± 0.70

(43.33) 56.67

Test group III- Methanol extract 250 mg/kg , p.o. 25 ± 1.29

(83.33) 16.67

Test group IV- Methanol Extract 500 mg/kg , p.o. 21 ± 1.13

(69.99) 30.01

Values are expressed as mean ± S.E.M.

indicates P < 0.001 vs. control; %: percentage; p.o.: per oral.

Table 2. Effect of T. orientalis on castor oil induced diarrhea in mice (Latent Period)

Animal group/Treatment Dose (mg/kg, per oral) Latent Period (h)

group-I, Control (1% tween-80) 10 1.096 ± 0.025

group-II, Standard (loperamide) 4 1.284 ± 0.053

group-III, Test group (Aqueous extract) 500 1.456 ± 0.029

group-IV (Methanol extract) 500 1.240 ± 0.058

Values are expressed as mean ± S.E.M. (n = 5) P < 0.025 vs control.

2008 Oriental Pharmacy and Experimental Medicine 8(2), 00-00

4 Sarder Nasir Uddin et al.

compared to the control and standard (Table 3).

During the first hour, the number of stool was 6.6

and 6.2 for aqueous and methanol extract respectively,

but the same time it was 7.8 and 7.2 for control and

standard (Table 3). The mean number of stools was

6.2, 4.4, 3.8, 1.2 and 0.6 at the 1

to 5

hour for

aqueous extract and 6.6, 3.8, 0.8, 0.8 and 0.2 at the

to 5

hour for methanol extract respectively. The

given data showed that the increase in period

decreases the number of stools.

DISCUSSION

For the millions of rural populations in countries of

the developing world, diarrhoeal diseases continue

to be the major cause of morbidity and mortality,

with an estimated 1 billion episodes of illness and

some 5 million or more deaths in children under

5 years. In such populations, preparations from

herbs and plants remain the most common forms

of treatment for diarrhoeal disease.

Acetic acid induced writhing model represents

pain sensation by triggering localized inflammatory

response. Acetic acid, which is used to induce

writhing, causes algesia by liberation of endogenous

substances, which in turn excite the pain nerve

endings (Taesotikul et al., 2003).

The acetic acid induced writhing demonstrates a

noxious stimulation in mice. The test consists of

injecting 0.7% acetic acid solution intraperitoneally

and then observed the animal for specific contraction

of body referred as ‘writhing’. A comparison of

writhing was made between positive control

(Diclofenac-Na), control and test sample given

orally 30 min prior to acetic acid injection. If the

sample possesses analgesic activity, the animal that

received the sample will give lower number of

writhing than the control, i.e. the sample having

analgesic activity will inhibit writhing.

From the study it was found that aqueous extract

of leaves at the doses of 500 mg/kg exhibited highly

significant (P < 0.001) inhibition of writhing reflex

by 56.76% while the standard drug diclofenac (25

mg/kg body weight) inhibition was found to be

48.34% (Table 1). So, it is concluded that the aqueous

extract of T. orientalis possess strong analgesic action.

Antidiarrhoeal activity of the aqueous and

methanol extract of Trema orientalis was tested

using the model of castor oil induced diarrhea in

mice. Castor oil, which was used to induce

diarrhea in mice, mixes with bile and pancreatic

enzymes and liberates ricinoleic acid from the

triglycerides upon oral administration. Most of the

ricinoleic acid remains in the intestine and

produces its anti absorptive effect. The ricinoleic

acid thus liberated readily forms ricinoleic salts

with sodium and potassium in the lumen of the

intestine. The salt formed as such behaves like a

soap or surfactant within gut and at the mucosal

surface. Most agreed view is that ricinoleate salt

Table 3. Effect of Trema orientalis on castor oil

induced diarrhea in mice (number of stools)

Animal

group/

Treatment

Dose

(mg/kg, per

oral)

Period of

study

(h)

Total no.

of stool

Control

(1% tween-80)

1 7.8 ± 0.73

2 5.0 ± 0.59

3 4.0 ± 0.63

4 3.6 ± 0.60

5 1.2 ± 0.48

Standard

(loperamide)

1 7.2 ± 0.73

2 4.6 ± 0.67

3 3.2 ± 0.37

4 1.28 ± 0.48

5 0.6 ± 0.40

Test group

(Aqueous

extract)

500

1 6.6 ± 0.6

2 3.8 ± 0.73

3 0.8 ± 0.48

4 0.8 ± 0.48

5 0.2 ± 0.2

Test group

(Methanol

extract)

500

1 6.2 ± 0.86

2 4.4 ± 0.5

3 3.8 ± 0.68

4 1.2 ± 0.37

5 0.6 ± 0.39

Values are expressed as mean ± S.E.M. (n = 5)

P < 0.025

vs control.

Analgesic and antidiarrhoeal activities of Treama orientalis Linn. in mice 5

2008 Oriental Pharmacy and Experimental Medicine 8(2), 00-00

stimulates the intestinal epithelial cell’s adenyl

cyclase (Racusen et al., 1979) or release prostaglandin

(Beubler et al., 1979).

The results showed that both the extract increase

the latent period and decrease the number of stools

i.e. delayed the onset of diarrhoeal episode at the

dose of 500 mg/kg body weight. If the sample

possesses antidiarrhoeal activity, the animal that

received the sample will give longer latent period

than the control, i.e. the sample having antidiarrhoeal

activity will increase the latent period and the

number of stools will decrease.

However, in toxicity evaluation of plant extracts

by brine shrimp lethality bioassay, aqueous extract

did not show any cytotoxic effect on brine shrimp

but methanol extract displayed toxicity. The

methanol extract showed cytotoxicity LC50 = 120 µg/

ml and LC

= 200 µg/ml. So, we have concluded

that the tested aqueous extract sample may be a

good source of analgesic and antidiarrhoeal medicine.

CONCLUSION

Bangladesh imports a large quantity of pharmaceutical

raw materials including medicinal plants and semi

processed plant products to produce drugs and

medicines. This huge foreign exchange can be

saved if the indigenous medicinal plants or their

semi-processed products are utilized by the

manufacturers to satisfy their needs. From the

above experiment, it could be suggested that both

aqueous and methanol extract of T. orientalis leaves

posses significant analgesic and antidiarrhoeal

activity in mice. So, further pharmacological and

toxicological study is required to establish the

therapeutic uses of the plant.

REFERENCES

Beubler E, Juan H. (1979) Effect of Ricinoleic acid and

other Laxatives in net water flux and prostaglandin

E release by the rat colon. J. Pharm. Pharmacol. 31,

681-685.

Chatterjee TK. (1993) Handbook of laboratory Mice

and Rats. pp. 133-139.

Farnsworth NR, Soejarto DD. (1991) Global importance

of medicinal plants. In: Akerele, O, Heywood V,

Synge H. (Eds.)

“The Conservation of Medicinal Plants”,

Cambridge University Press, Cambridge, 25-51.

Iwe M M. (1993) Hand Book of Medicinal Plants. CRC

Publication. p. 251.

Kirtikar, KR. and Basu, BD. 1980. Indian Medicinal

Plants. 2nd Ed, Vol. 1. p. 264.

Racusen LC, Binder HJ. (1979) Ricinoleic acid stimulation

of active anion secretion in colonic mucosa of the

rat. J. Clin. Invest. 63, 743-749.

Rulangaranga ZK. (1991) Conservation of medicinal

and aromatic plants in Tanzania, proceedings of a

workshop on priority species for tree planting and

afforestation in Tanzania, Morogoro, Tanzania.

May 14-18, 1990.

Taesotikul T, Panthong A, Kanjanapothi D, Verpoorte

R, scheffer JJC. (2003) Anti-inflammatory, antipyretic

and antinociceptive activities of Tabernaemontana

pandacaqui Poir. J. Ethnopharmacol. 84, 31-35.

Whittle BA. (1964) The use of changes in capillary

permeability in mice to distinguish between narcotic

and non-narcotic analgesics. Br. J. Pharmacol. Chemother.

22, 246-253.

World Health Organization. (1994) The treatment and

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WHO, Geneva. (2nd Ed) pp. 1-4.

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Chapter

Mar 2025

Underutilized Wild Edible Fruits is a treasure trove of minor, neglected and often overlooked fruits that have played significant roles in human diets for centuries. These fruits, characterized by their local, orphaned, and promising nature, have faced challenges such as low supply, poor shelf life, and limited consumer awareness, leading to their diminished importance in modern times. Despite their underutilization, wild edible fruits have been vital sources of sustenance, flavour and even medicinal properties throughout history. They offer alternative food sources during periods of scarcity, potential nutritional supplements to combat malnutrition and income opportunities for rural communities. However, with the dominance of industrial agriculture and globalization, traditional knowledge of these fruits has dwindled and commercially cultivated varieties have overshadowed them. Yet, there is a growing interest in reviving and exploring these underexploited fruits. Researchers, nutritionists, and conservationists are recognizing their nutritional richness, potential for biodiversity conservation, and contributions to sustainable development. This chapter aims to provide a comprehensive overview of underexploited wild edible fruits, delving into their nutritional composition, culinary uses, traditional knowledge, conservation status and potential for cultivation. By shedding light on these often-neglected fruits, we hope to cultivate a deeper appreciation for biodiversity and advocate for sustainable methodologies to preserve these invaluable resources for future generations.

Global Importance of Medicinal Plants

Chapter

Jul 1991

Nearly all cultures, from ancient times to today, have used plants as a source of medicine. In many developing countries, traditional medicine is still the mainstay of health care and most of the drugs and cures used come from plants. In developed countries many people are turning to herbal remedies. With this widespread use has come the assumption that plants identified as having medicinal qualities will be available on a continuing basis. However no concerted effort has been made to ensure this and in the face of the threats of increasing demand, a vastly increasing human population and extensive forest destruction, there can be no guarantee that we will continue to benefit indefinitely from this valuable resource. In light of this situation the World Health Organisation held a meeting in 1988. This book is the outcome of that meeting, detailing in a series of papers by leading experts the problems of which need to be addressed, the existing experiences from a range of countries and the future direction which must be taken to ensure the conservation of the world's medicinal plants.

Illustrated Indian Medicinal plants III

Article

Jan 2000

Handbook on Laboratory Mice and Rats

Article

Jan 1993

TK. Chatterjee

Effect of ricinoleic acid and other laxatives on net water flux and prostaglandin E release by rat colon

Article

Nov 1979

Ricinoleic acid, oleic acid, dioctyl sodium sulphosuccinate, deoxycholic acid, sennoside A + B and mannitol reduced or reversed water flux from lumen to blood in rat colon in situ. Stearinic acid was without any effect. Ricinoleic acid, oleic acid, dioctyl sodium sulphosuccinate, deoxycholic acid and sennoside A + B stimulated release of PGE-like material into the colonic lumen whereas the osmotic laxative mannitol and stearinic acid did not. Inhibition of PGE biosynthesis by pretreatment of the rats with indomethacin significantly reduced (but did not abolish) the effect of ricinoleic, oleic and deoxycholic acids on net water flux and PGE release. Indomethacin reduced the effect of dioctyl sodium sulphosuccinate and of sennoside A + B on PGE release but not their effect on the net water flux. The effect of mannitol was not influenced by indomethacin. The amount of PGE release in experiments with ricinoleic acid, oleic acid, stearinic acid and dioctyl sodium sulphosuccinate (with and without indomethacin) showed a good correlation (r = 0.99) with the change in net water flux. Deoxycholic acid, sennoside A + B and mannitol did not show this correlation. It is assumed that the action of non-osmotic laxatives is partially mediated by PGE, although other mechanisms also seem to be involved in their mode of action.

Ricinoleicacid stimulation of active anion secretion in colonic mucosa of the rat

Article

May 1979

Perfusion of the colon with ricinoleic acid produces fluid and electrolyte accumulation. The mechanism of these changes in water and electrolyte movement is uknown. These studies were designed to determine whether ricinoleic acid effects active ion transport across isolated rat colonic mucosa. 0.5 mM Na ricinoleate produced significant increases in potential difference (3.8 +/- 0.5 mV) and short-circuit current (Isc) (99.2 +/- 10.1 muA/cm2). The increases in Isc produced by Na ricinoleate were inhibited by both removal of bicarbonate and chloride and by the presence of theophylline. The hydroxy fatty acid also resulted in a significant decrease in net Na absorption from 4.7 +/- 0.8 to 0.1 +/- 0.7 mueq/h cm2 and reversed net Cl transport from absorption (+ 4.5 +/- 0.9) to secretion (-2.2 +/- mueq/h cm2). In parallel studies 0.5 mM Na ricinoleate increased mucosal cyclic AMP content by 58%. The concentrations of Na ricinoleate required to produce detectable and maximal increases in both Isc and cyclic AMP were the same. These results provide evidence in support of the concept that hydroxy fatty acid-induced fluid and electrolyte accumulation is driven by an active ion secretory process.

Anti-inflammatory, antipyretic and antinociceptive activities of Tabernaemontana pandacaqui Poir

Article

Feb 2003
J ETHNOPHARMACOL

Studies on carrageenin-induced rat paw edema, yeast-induced hyperthermia in rat and writhing response induced by acetic acid in mice showed that the alcoholic extract of stems of Tabernaemontana pandacaqui (T. pandacaqui) has significant anti-inflammatory, antipyretic and antinociceptive activities. These activities are due to alkaloidal components since they were also observed when the crude alkaloidal (CA) fraction separated from alcoholic extract was tested in the same models.

The use of changes in capillary in mice to distinguish between narcotic and non-narcotic analgesia

Article

May 1964
Br J Pharmacol Chemother

BA Whittle

An extension of the "squirming test" is described which makes the method specific for nonnarcotic analgesics. The intraperitoneal injection of acetic acid causes squirming and an increase in capillary permeability that is measured by direct estimation of plasma-bound dye (Pontamine Sky Blue) which has leaked into the peritoneal cavity. Nonnarcotic analgesics inhibit squirming and leakage of dye. Values for the oral ED50s for both effects are given for a number of typical compounds. Narcotic analgesics, in doses that produce analgesia, inhibit squirming but do not significantly affect leakage of dye. Drugs that stimulate the central nervous system and also inhibit squirming have no significant effect on leakage of dye over the range of doses which inhibit squirming. Corticosteroids do not significantly inhibit either squirming or leakage of dye.

The treatment and prevention of acute diarhoea, Practical Guidelines. WHO, Geneva

Jan 1994
1-4

World Health Organization. (1994) The treatment and prevention of acute diarhoea, Practical Guidelines. WHO, Geneva. (2nd Ed) pp. 1-4.

Hand Book of Medicinal Plants

Jan 1993
251

M M Iwe

Iwe M M. (1993) Hand Book of Medicinal Plants. CRC Publication. p. 251.

Conservation of medicinal and aromatic plants in Tanzania

Jan 1990

Z K Rulangaranga

Rulangaranga ZK. (1991) Conservation of medicinal and aromatic plants in Tanzania, proceedings of a workshop on priority species for tree planting and afforestation in Tanzania, Morogoro, Tanzania. May 14-18, 1990.