Phytomedicine 11: 315322, 2004

http://www.elsevier.de/phymed

Analgesic and anti-inflammatory activities

of a fraction rich in oncocalyxone A isolated
from Auxemma oncocalyx
M. A. D. Ferreira1, Osmar D. R. H. Nunes1, Juvenia B. Fontenele3, Otlia D. L. Pessoa2,
Telma L. G. Lemos2 and Glauce S. B. Viana3
1
Departamento de Farmcia
2
Departamento de Qumica Orgnica e Inorgnica
3
Departamento de Fisiologia e Farmacologia Universidade Federal do Cear, Fortaleza Ce Brasil

Summary
In the present work we studied the antinociceptive and antiedematogenic effects of a quinone frac-
tion (QF) isolated from the heartwood of Auxemma oncocalyx Taub. The major constituent of QF,
which represented around 80% of this fraction, was a terpenoid quinone named oncocalyxone A
(1). Results show that QF (10 and 30 mg/kg body wt., i.p.) significantly inhibited paw edema in-
duced by carrageenan at the second, third, and fourth hours. The effect was dose-dependent and
long lasting, and QF was less effective orally. An antiedematogenic effect was also demonstrated in
the dextran-induced paw edema. In this model, however, QF was somewhat less potent. QF (1 and
5 mg/kg body wt., i.p.) inhibited acetic acid-induced abdominal contractions in mice in a dose-de-
pendent manner. In addition, QF (5 and 10 mg/kg body wt., i.p.) inhibited only the second phase
(inflammatory) in the formalin test, and showed no effect in the hot-plate test in mice. The
antinociceptive activity of QF was predominantly peripheral and independent of the opioid sys-
tem. The observed effects of QF are, at least in part, probably due to the presence of oncocalyxone
A (1).

Key words: Analgesic and antiinflammatory activities, oncocalyxone A, Auxemma oncocalyx, quinones

Introduction
Auxemma oncocalyx Taub. belongs to the Boragi-
naceae family and is native to the northeastern Brazil-
ian caatinga, where it is known as pau branco. The
stem bark of the tree is astringent and used popularly in
the treatment of wounds (Braga, 1976; Pessoa, 1994).
Alantoin isolated from the stem hydroalcoholic extract
(Pessoa and Lemos, 1997) explains partially the bene-
ficial use of the plant in wound healing. Several other
compounds were also isolated from the hydroalcoholic
extract, including -sitosterol, its glycoside (3-O-D-
glucopyranosylsitosterol) and seven terpenoidic
quinones, among them oncocalyxone A, 1 (Pessoa
et al. 1993; Pessoa et al. 1995).
Previous pharmacological studies showed that the
stem hydroalcoholic extract possesses anti-platelet ac-
tivity (Fontenele and Sousa, 1992) and this effect was
confirmed recently in a hydrosoluble fraction, obtained
from the heartwood methanolic extract, which also pre-
sented antioxidant activity (Ferreira et al. 1999; Fer-
reira et al. 2001). Other effects were also attributed to
the hydroalcoholic extract, such as antitumoral (Pessoa
et al. 1992), analgesic and anti-inflammatory (Lino

0944-7113/04/11/04-315 $ 30.00/0
316 M. A. D. Ferreira et al.
et al. 1996) activities. Recently, in vitro studies demon-
strated the antitumoral activity not only of oncocalyx-
one A (1), but also of oncocalyxone C, isolated from A.
oncocalyx (Pessoa et al. 2000; Leyva et al. 2000).
Quinones show several types of biological activity,
including, platelet antiaggregant (Teng et al. 1993;
Chung et al. 1994), antioxidant (Belisario et al. 1992;
Houghton et al. 1995; Tripathi et al. 1995; Mori et al.
1998), anti-inflammatory (Kuo et al. 1995; Vazquez et
al. 1996; Odukoya et al. 1999), analgesic (Hernndez-
Perez et al. 1995; Abdel-Fattah et al. 2000), antitumoral
(Morello et al. 1995; Itoigawa et al. 2000), antifungal
(Perry et al. 1991; Gafner et al. 1996), antimalarial
(Figueiredo et al. 1998), and leishmanicidal (Sauvain et
al. 1993; Sittie et al. 1999). The objectives of the present
work were to study the analgesic and anti-inflammatory
effects of the quinone-enriched fraction from A. oncoca-
lyx and to elucidate its possible mechanism of action.

Materials and Methods
Plant material and fractionation
The plant was collected at the city of Pentecoste, state
of Cear, in northeastern Brazil, and identified by Prof.
A. G. Fernandes of the Biology Department of the Fed-
eral University of Cear. A voucher specimen (No.
18459) has been deposited at the Prisco Bezerra
Herbarium. The quinone fraction (QF) was prepared
from the ground heartwood methanolic extract through
exhaustive aqueous extraction followed by lyophiliza-
tion. The hydrosoluble fraction contained around 80%
of oncocalyxone A (1) (Fig. 1), which was previously
characterized by spectroscopy analysis as MS, IR and
NMR including 2D sequences such as COSY, HET-
COR and COLOC (Pessoa et al. 1993). The concentra-
tion of (1) in the hydrosoluble fraction was calculated
on the basis of the HPLC-fingerprint analysis using
YMC-Pack C-18 column, acetonitrile :H2O (1:1, v/v)
as mobile phase with an isocratic elution program at
room temperature and detection at 280 nm.
Fig. 1. Chemical structure of Oncocalyxone A (Pessoa et al,
1993).
Test animals
Male Wistar rats (150200 g each) and male Swiss mice
(2530 g each) were obtained from the Animal House
of the Federal University of Cear. Animals were main-
tained in an air-conditioned room at 2325 C, 12 h
light-12 h dark cycle, and fed with a standard laboratory
diet and tap water ad libitum. Experiments were per-
formed according to the Guide for use and care of labo-
ratory animals of the Department of Health and Human
Services of the United States of America.
Drugs
Carrageenan, dextran and naloxone were purchased
from Sigma, U.S.A. Morphine sulfate was obtained
from Cristlia, Brazil. All other drugs were of analyti-
cal grade.
Pharmacological tests
Carrageenan-induced paw edema (Winter et al. 1962):
Male Wistar rats (150200 g each) divided into groups
of 6 animals each were used. QF was administered at
doses of 1, 10 and 30 mg/kg body wt., i.p., 30 min be-
fore the intraplantar injection of a 1% carrageenan so-
lution (0.1 ml in the right hind paw). Paw volume was
measured using a plethysmometer (Ugo Basile, Italy),
before and at 1; 2; 3; 4 and 24 h after carrageenan ad-
ministration. The difference between the paw volume
before and after carrageenan administration was con-
sidered as an index of edema formation at each obser-
vation time. Controls received saline (10 ml/kg body
wt., i.p.). QF (100 and 200 mg/kg body wt.) was also
administered orally, 60 min before injection of car-
rageenan, and in this case controls received a 2%
DMSO solution in saline (used as vehicle for QF) and
animals were fasted for approximately 12 h.
Dextran-induced paw edema (Parrat and West,
1958): Male Wistar rats (150200 g each) divided into
6 animals per group were used. QF was administered at
doses of 1, 10 and 30 mg/kg body wt., i.p., 30 min be-
fore the intraplantar injection of 0.1 ml of 1.5% dextran
solution in the right hind paw. The paw volume was
measured using a plethysmometer before and 1/2; 1; 2; 3
and 4 h after dextran injection. The difference between
the paw volume before and after dextran injection is
considered as an index of edema at each observation
time. Controls received saline (10 ml/kg body wt., i.p.).
Acetic acid-induced abdominal contractions (Koster
et al. 1959): Male Swiss mice (2030 g each), divided
into groups of 10 animals each were used. The quinone
fraction (QF) was administered once at doses of 0.1,
1 and 5 mg/kg body wt., i.p., 30 min before the 0.6%
acetic acid injection (10 ml/kg body wt., i.p.). Ten min-
utes later, the number of abdominal contractions was
registered for 20 min. The control group received
saline, used as vehicle for QF.

Formalin test (Hunskaar et al. 1985; Tjolsen et al.
1992): Male Swiss mice (2030 g each) divided into
groups of 6 animals each were used. QF was adminis-
tered once at doses of 1, 5, 10 and 30 mg/kg body wt.,
i.p., 30 min before the intraplantar injection of 20 l of
1% formalin in the right hind paw. The licking time (in
seconds) was registered at 5 min (first phase, neuro-
genic), and after 20 min for 5 min (second phase, in-
flammatory), after formalin injection. The control
group received saline, 10 ml/kg body wt., i.p. Mor-
phine was used as a standard (5 mg/kg body wt., i.p.).
In order to verify the possible involvement of the opi-
oid system in the analgesic effect of QF, the formalin
test was performed also in the presence of the opioid
antagonist, naloxone (2 mg/kg body wt., subcutaneous-
ly). QF (30 mg/kg body wt., i.p.) and morphine were
administered 15 min after and 30 min before naloxone
and formalin injections, respectively.
Hot-plate test (Eddy and Leimback, 1953): Male
Swiss mice (2030 g each) divided into groups of 10
animals each were used. Animals were selected previ-
ously from those presenting a latency to the thermal
stimulus equal to or less than 20 sec; and the cut off
point was set at 40 sec. QF was administered once at
doses of 5, 10 and 30 mg/kg body wt., i.p. Latency to
the thermal stimulus was registered before and at 30,
60 and 90 min after QF administration. The control
Analgesic and anti-inflammatory activities 317
group received saline (10 ml/kg body wt., i.p.). Mor-
phine (5 mg/kg body wt., i.p.) was used as a standard.
Statistical analysis
Data were analyzed by one-way Analysis of Variance
(ANOVA) followed by the Tukey-Kramer test for mul-
tiple comparisons.

Results
The quinone fraction was very active in reducing the rat
paw edema after carrageenan administration (Table 1).
The results show inhibitions of approximately 46, 57
and 48%, at the second, third and fourth hours, respec-
tively, after the dose of 10 mg/kg body wt., i.p. and of
50, 60 and 51% at the same intervals following the
dose of 30 mg/kg body wt., i.p. The effect was long-
lasting and reductions of 38% in the edema could still
be seen after a 24-h period (with the highest dose).
Although less potent, QF was also effective after oral
administration (100 and 200 mg/kg body wt.); howev-
er, under these conditions, decreases of edema ranging
from 25 to 29% were observed at the third, fourth and
twenty-fourth hours after carrageenan administration,
as compared to controls.

Table 1. Effect of the quinone fraction (QF) of Auxemma oncocalyx Taub. on carrageenan-induced paw edema in rats.

Group (n) Edema volume in ml (% Inhibition)

1h 2h 3h 4h 24 h

Control, i.p. (6) 0.61 0.03 1.03 0.10 1.62 0.10 1.41 0.13 0.29 0.04
QF
1 mg/kg body wt., i.p. (6) 0.59 0.06 0.98 0.09 1.35 0.17 1.28 0.11 0.27 0.03
(3.3) (4.9) (16.7) (9.2) (6.9)
10 mg/kg body wt., i.p. (6) 0.45 0.05 0.56 0.05* 0.69 0.06* 0.74 0.06* 0.20 0.02
(26.2) (45.6) (57.4) (47.5) (31.0)
30 mg/kg body wt., i.p. (6) 0.44 0.04 0.52 0.05* 0.65 0.05* 0.69 0.05* 0.18 0.02
(27.9) (49.5) (59.9) (51.0) (37.9)

Control, p.o. (6) 0.94 0.06 1.54 0.13 2.06 0.10 2.09 0.11 0.24 0.04
QF
100 mg/kg body wt., p.o. (6) 0.79 0.14 1.57 0.08 1.70 0.13 1.93 0.09 0.19 0.02
(15.9) () (17.5) (7.7) (20.8)
200 mg/kg body wt., p.o. (6) 0.73 0.06 1.47 0.18 1.54 0.16* 1.58 0.12* 0.17 0.02
(22.3) (4.6) (25.2) (24.4) (29.2)

Male Wistar rats (150200 g each) were used. Control animals received saline (10 ml/kg body wt.) and treated groups received
QF intraperitoneally or orally, 30 or 60 min, respectively, before the intraplantar administration of 1% carrageenan solution
(0.1 ml, right hind paw). Edema volume (expressed in ml) was estimated at 1, 2, 3, 4 and 24 h after carrageenan administration.
Values are expressed as means S.E.M. of the number of animals. *p < 0.05 vs. control (One-way ANOVA and Tukey-Kramer
as post-hoc test).
318 M. A. D. Ferreira et al.
Table 2. Effect of the quinone fraction (QF) of Auxemma oncocalyx Taub. on dextran-induced paw edema in rats.

Group (n) Edema volume in ml (% Inhibition)

1
/2 h 1h 2h 3h 4h

Control, i.p. (6) 2.4 0.13 3.0 0.13 3.1 0.11 3.1 0.08 2.4 0.15
QF 1 mg/kg body wt., i.p. (6) 2.8 0.21 3.1 0.18 3.0 0.12 2.7 0.14 2.7 0.12
QF 10 mg/kg body wt., i.p. 2.2 0.11 2.4 0.08 2.0 0.24* 1.8 0.24* 1.6 0.25
(8.3) (20.0) (35.5) (41.9) (33.3)
QF 30 mg/kg body wt., i.p. (6) 2.0 0.30 2.2 0.27* 1.7 0.23* 1.6 0.28* 1.4 0.24*
(16.7) (26.7) (45.2) (48.4) (41.7)

Male Wistar rats (150200 g each) were used. Control animals received saline (10 ml/kg body wt.) and treated groups received
QF 30 min before the intraplantar injection of 0.1 ml, 1.5% dextran solution in the right hind paw. Edema volume (in ml) was
estimated at 1/2, 1, 2, 3 and 4 h after dextran administration. Values are expressed as means S.E.M. of the number of animals.
*p < 0.05 vs. control (One-way ANOVA and Tukey-Kramer as post-hoc test).

Table 3. Antinociceptive effect of the quinone fraction (QF)
of Auxemma oncocalyx Taub. on acetic acid-induced abdomi-
nal contractions in mice.
Group (n) Number of % Inhi-
contractions bition
Control, i.p. (16) 24.5 1.80
QF 0.1 mg/kg body wt., i.p. (10) 23.6 1.09
QF 1 mg/kg body wt., i.p. (10) 18.2 1.42* 26
QF 5 mg/kg body wt., i.p. (10) 9.7 0.92* 60
Male Swiss mice (2030 g each) were used. Control animals
received saline (10 ml/kg body wt.) and treated groups re-
ceived QF 30 min before an 0.6% acetic acid injection
(10 ml/kg body wt., i.p.). Ten minutes later, the number of ab-
dominal contractions was registered over 20 min. Values are
expressed as means S.E.M. of the number of animals used
in the test (in parentheses). *p < 0.05 vs. control (One-way
ANOVA and Tukey-Kramer as post-hoc test). Percentages of
inhibition of contraction number relative to control are also
presented.
Data in Table 2 show the effect of QF (1, 10 and
30 mg/kg body wt., i.p.) on dextran-induced paw
edema in rats. Significant reductions of paw edema
were observed at the dose of 30 mg/kg body wt., i.p., at
the first (27%), second (45%), third (48%) and fourth
(42%) hours. QF was active at the dose of 10 mg/kg
body wt., i.p., only at the second (36%), third (42%)
and fourth (33%) hours.
Table 3 shows the effects of QF on acetic acid-induced
abdominal contractions in mice. QF (1 and 5 mg/kg body
wt., i.p.) inhibited abdominal contractions by 26 and
60%, respectively, as compared to controls (24.5 1.8
contractions/20 min). No effect was observed, however,
at the lower dose of QF (0.1 mg/kg body wt., i.p.).
Table 4 shows the effects of QF on formalin-induced
nociception in mice. QF at doses of 5, 10 and 30 mg/kg
body wt., i.p., significantly inhibited licking time in the
second phase of the response by 27, 52 and 62%, re-
spectively, as compared to controls. No effect was ob-
served in the first phase of the response. On the other
hand, morphine inhibited licking time in the first and
second phases of the response by 55 and 85%, respec-
tively. Although the effects of morphine were totally
reversed, no change was seen in the effect of QF in the
presence of naloxone, as compared to the effect of QF
alone.
QF at doses of 5, 10 and 30 mg/kg body wt., i.p. did
not, however, alter latency of reaction to the thermal
stimulus, as demonstrated by the hot-plate test. Mor-
phine, used as positive control, produced increases on
the order of 84, 57 and 65% after 30, 60 and 90 min, re-
spectively, as compared to control (Table 5).

Discussion
The present work showed that QF, administered in-
traperitoneally or orally, inhibited carrageenan-induced
rat paw edema in a dose-dependent manner. The maxi-
mum effect was observed even at the dose of 10 mg/kg
body wt., i.p. The inhibition was long-lasting, still ob-
servable 24 h after carrageenan administration. The in-
hibition was significant at the second, third and fourth
hours, when bradykinin is being released and there oc-
curs an accumulation of prostaglandins and infiltration
of polymorphonuclear cells. At this stage, there is also
production of free radicals, among other mediators.
The antiedematogenic effect of QF was greater after in-
traperitoneal administration, indicating that oral ab-
sorption is less efficient. This effect was also seen in
 Analgesic and anti-inflammatory activities 319
Table 4. Evaluation of the opioid systems involvement in the effect of the quinone fraction (QF) of Auxemma oncocalyx
Taub. on formalin-induced nociception in mice.

Group Licking time (sec) % Inhibition

first phase second phase first phase second phase

Control, i.p. (21) 56.6 2.57 26.4 1.86

QF 1 mg/kg body wt. (11) 55.1 3.06 25.3 1.26
QF 5 mg/kg body wt. (12) 53.5 3.14 19.3 1.68* 27
QF 10 mg/kg body wt. (10) 58.5 2.73 12.8 1.22* 52
Control, i.p. (12) 50.9 2.63 28.2 2.77
QF 30 mg/kg body wt. (11) 48.4 3.24 10.7 1.35* 62
Mor (12) 22.9 1.75* 4.2 0.89* 55 85
Nal + QF (10) 47.4 3.34 9.1 1.57* 68
Nal + Mor (12) 50.6 4.41 25.6 2.69

Male Swiss mice (2030 g each) were used. Control group received saline (10 ml/kg body wt.) and treated groups received QF
or morphine (5 mg/kg body wt.) intraperitoneally, 30 min before the intraplantar injection of 20 l of 1% formalin in the right
hind paw. The licking time (in seconds) was registered at the first 5 min (first phase) and after 20 min over 5 min (second
phase) after the formalin injection. In the test using Naloxone (Nal) and the quinone fraction, QF was administered 15 min
after naloxone (2 mg/kg body wt., subcutaneously) and 30 min before formalin injection. Morphine was also administered 15
min after Naloxone (Nal) and 30 min before formalin injection. Values are expressed as means S.E.M. of the number of ani-
mals in parentheses. *p < 0.05 vs. control (One-way ANOVA and Tukey-Kramer as post-hoc test). Percentages of inhibition of
licking time relative to control are also presented.

Table 5. Effect of the quinone fraction (QF) of Auxemma oncocalyx Taub. on the hot-plate test in mice.

Group Latency time (sec)

0 min 30 min 60 min 90 min

Control, i.p. (16) 15.8 1.13 15.2 0.96 13.5 0.87 12.6 1.10
QF 5 mg/kg body wt., i.p. (10) 13.6 1.38 14.2 1.06 10.9 0.57 13.3 1.28
QF 10 mg/kg body wt., i.p.(10) 12.5 1.34 13.7 1.74 11.2 1.37 11.8 1.16
QF 30 mg/kg body wt., i.p. (10) 16.0 1.51 16.2 1.04 15.8 0.87 12.6 1.29
Morphine 5 mg/kg body wt., i.p. (10) 15.3 1.54 28.0 3.04* 23.7 2.09* 19.3 1.68*

Male Swiss mice (2030 g each) were used. Control animals received saline (10 ml/kg body wt.) and treated groups received
QF or morphine. The latency to the thermal stimulus was registered (in seconds) before (0 min) and at 30, 60 and 90 min after
QF administration. Values are expressed as means S.E.M. of the number of animals in parentheses. *p < 0.01 vs. control
(One-way ANOVA and Tukey-Kramer as post-hoc test).

the dextran-induced paw edema test, where QF inhibit-
ed the edema dose-dependently after intraperitoneal
administrations of 10 and 30 mg/kg body wt.
Earlier research showed that in carrageenan-induced
edema, several mediators (histamine, 5-hydroxytrypt-
amine, kinins, prostaglandins) of the inflammatory
process are involved, including the complement sys-
tem (Willis, 1969; Willoughby et al. 1969). Di Rosa
et al. (1971) showed that this system participates in the
process from the very beginning, and that carrageenan-
induced inflammation is divided into 3 steps according
to the mediators released. In the initial step (first
90 min), the release of both histamine and 5-hydroxy-
tryptamine (5-HT) occurs; and the second step (from
90 to 150 min) is mediated by kinins, while in the third
step (from 150 min on), prostaglandin release takes
place. All these mediators are dependent upon the com-
plement system. Histamine and 5-HT are responsible
for vasodilation and increase in vascular permeability
in the initial phase of the inflammatory process.
Bradykinin has been implicated in acute inflammatory
processes due to its ability to induce an increase in
blood vessel permeability. Its involvement in car-
rageenan-induced edema was demonstrated by Ronald
and Christopher (1990) and by Damas and Remacle-
Volon (1992). Another characteristic of carrageenan-
320 M. A. D. Ferreira et al.
induced edema is the massive infiltration of polymor-
phonuclear leucocytes observed in the third step (Di
Rosa and Willoughby, 1971; Ialenti et al. 1992; Masso
et al. 1993). By contrast, dextran-induced edema in-
volves mast cell degranulation, resulting in histamine
as well as 5-HT release.
Histamine, 5-HT and bradykinin are able to induce
nitric oxide (NO) release from vascular endothelial
cells in vitro via a mechanism involving receptor occu-
pation and stimulation of NO synthase. In addition,
macrophages produce NO when activated by
lipopolysaccharide or cytokines. Thus Salvemini et al.
(1996) pointed to NO as an important mediator of car-
rageenan-induced edema and suggested that constitu-
tive NO synthase (cNOS) acts at the initial steps, while
inducible NO synthase (iNOS) is involved in the last
step of the inflammatory reaction.
The acute or chronic inflammatory process is a
model of nociception (Besson, 1997) and several
works have shown that inflammatory mediators can
generate nociceptive impulses (Steen et al. 1996;
Besson, 1997). Thus, bradykinin and 5-HT are able to
stimulate cutaneous nociceptors and 5-HT can also in-
crease nociceptive response to bradykinin (Lang et al.
1990; Rueff and Dray, 1993). Although prostaglandins
are not able to activate these nociceptors, they promote
sensitization of these same nociceptors to bradykinin in
some tissues (Schaible and Schmidt, 1988). Addition-
ally, neuropeptides such as substance P play a role in
neurogenic inflammation (Steen et al. 1996). Recent
evidence showed the importance of cytokines, includ-
ing tumor necrosis factor (TNF), in the generation
and maintenance of hyperalgesia associated with neu-
ropathic pain (Jungler and Sorkin, 2000).
Considering the relationship between anti-inflam-
matory and analgesic effects, another objective of the
present work was to study the antinociceptive activity
of QF using the writhing and formalin test in mice.
Acetic acid produces a painful reaction and acute in-
flammation in the peritoneal area. The stimulation of
peritoneal nociceptors is indirect and occurs through
the release of endogenous substances, which stimulate
nervous endings (Gyires and Torna, 1984). A great in-
crease occurs in prostaglandins E2 and F2a levels in
peritoneal fluid after acetic acid injection, and the anal-
gesic effect of substances similar to aspirin could be
due to the blockade of prostaglandin synthesis.
QF was able to significantly and dose-dependently
inhibit the abdominal contractions induced by acetic
acid, as well as the second phase of the response to for-
malin. The formalin test is considered a model for
chronic pain (Dubuisson and Dennis, 1977). In this
test, animals present two distinct nociceptive behavior
phases, which probably involve different stimuli. The
first phase initiates immediately after formalin injec-
tion and lasts 3 to 5 min, resulting from chemical stim-
ulation of nociceptors. The second phase initiates 15 to
20 min after formalin injection, lasts 20 to 40 min and
seems to depend on a peripheral mechanism as well as
a central one. While substance P and bradykinin are in-
volved in the first phase, histamine, 5-HT,
prostaglandins and bradykinin are involved in the sec-
ond phase. The effect of QF was significant only in the
second phase, indicating an action related to the in-
flammatory process. This QF effect was not reversed
by naloxone, pointing to the non-involvement of the
opioid system. Finally, QF showed no effect in the hot-
plate test, which involves higher brain functions and
consists of responses to nociceptive stimuli organized
at a supraspinal level (Gardmark et al. 1998). Analgesic
drugs such as aspirin and paracetamol do not have any
effect in this test (Tjolsen et al. 1992), while it is large-
ly used to measure opioid effects (Gong et al. 1991;
Plone et al. 1996).
Acknowledgements
The authors wish to thank the Brazilian National Research
Council (CNPq) for supporting this work and Maria V. R.
Bastos for technical assistance.

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Address
G. S. B. Viana, Departamento de Fisiologia e Farma-
cologia, Faculdade de Medicina, Universidade Federal
do Cear, Rua Barbosa de Freitas, 130/1100, CEP:
60170-020 Fortaleza Cear Brazil
Fax: 55 (85) 288-8333;
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