Pharmaceutical Biology, 2012; 50(1): 113–119

© 2012 Informa Healthcare USA, Inc.

ISSN 1388-0209 print/ISSN 1744-5116 online
DOI: 10.3109/13880209.2011.634423

Research Article

Sedative effects of essential oils obtained from

Baccharis uncinella
J. Ascari1, S.L. Sens2, D.S. Nunes1, A. Wisniewski Jr.3, M.D. Arbo4, V.M. Linck5,
P. Lunardi5, M.B. Leal4, and E. Elisabetsky5
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1
Universidade Estadual de Ponta Grossa, Mestrado em Química Aplicada, Ponta Grossa, PR, Brazil, 2Centro de
Educação Tecnológica Irmão Mário Cristóvão, Curitiba, PR, Brazil, 3Universidade Federal de Sergipe, Departamento
de Química, São Cristóvão, SE, Brazil, 4Laboratório de Farmacologia e Toxicologia de Produtos Naturais, Depto
de Farmacologia, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil, and 5Laboratório de
Etnofarmacologia, Depto de Farmacologia, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

Abstract
Context: Essential oils (EOs) have been reported to possess pharmacological properties, of which those related to
the central nervous system have been especially attributed to mono- and sesquiterpenes. Baccharis uncinella DC.
(Asteraceae) is used by the Laklaño Indians (Santa Catarina, Brazil) for sedative purposes. Interestingly, the species
For personal use only.

does not seem to be used medicinally elsewhere in Brazil.

Objective: This study was designed to compare the composition and sedative properties of B. uncinella EOs obtained
closer (BU-SC) and farther (BU-PR) to the Laklaño Indian Reserve.
Materials and methods: BU-SC and BU-PR obtained by hydrodistillation were analyzed by CG-MS. Mice treated
with BU-SC and BU-PR (50 and 100 mg/kg) were evaluated regarding pentobarbital-induced sleeping time, body
temperature, and locomotion.
Results: BU-SC presents a higher monoterpene/sesquitherpene ratio (0.31); α-pinene (6.42%), limonene (7.21%),
caryophyllene (26.13%), spathulenol (13.39%) and caryophyllene oxide (13.26%) were identified as major components.
BU-PR presents a low monoterpene/sesquitepene ratio (0.004); spathulenol (32.93%), caryophyllene oxide (27.78%),
viridiflorol (5.29%) and α-cadinol (2.42%) were identified as the main components. Both samples significantly (p < 0.05,
ANOVA) decreased locomotion and body temperature, as well as increased sleeping time. The hypnotic activity was
sensitive to the differences in monoterpene composition.
Conclusions: In comparison with a sample collected in Paraná State, B. uncinella EO collected closer to the Laklaño
Indians possess a composition that better justifies the claimed sedative properties. The study confirms the value of
traditional information to guide bioactivity assessment in medicinal plants, and gives notice to the ecological factors
that can interfere with the conclusions of such assessments.
Keywords:  Ethnopharmacology, Laklaño Indians, monoterpenes, sesquitepenes, hypno-sedative

Introduction
Medicinal plants present a variety of chemical constitu-
ents accumulated as byproducts of the plants secondary
metabolism. Among these chemicals, a distinct class
is composed by essential oils (EOs), of which some are
used in indigenous medical systems to treat various con-
ditions including mental illnesses (Umezu et  al., 2001,
2006). From the dozens of components usually found in
EOs, the monoterpenes and sesquiterpenes seem to be
the main responsible for central nervous system (CNS)
effects (Heuberger et al., 2010). For instance, the monot-
erpene (R)-(-)-linalool is recognized as the sedative/
calming component of numerous traditional and com-
mercial plant preparations and/or their EOs (Elisabetsky
et  al., 1995; Sugawara et  al., 1998; Kuroda et  al., 2005;
Shaw et al., 2007; Linck et al., 2009, 2010; Heuberger et al.,

Address for Correspondence:  Elaine Elisabetsky, Laboratório de Etnofarmacologia, Depto de Farmacologia, ICBS, Universidade Federal do
Rio Grande do Sul, Rua Sarmento Leite 500/202, Porto Alegre, RS, Brazil. Tel/Fax: 55 51 33083121. E-mail: elaine.elisabetsky@gmail.com
(Received 28 June 2011; revised 06 October 2011; accepted 19 October 2011)

113
 114  J. Ascari et al.
2010). α-Terpineol is another monoterpenoid derivative
identified as the active component of traditional seda-
tives (de Sousa et al., 2007).
The phytochemical analysis of many Baccharis spe-
cies indicates the presence of flavonoids, diterpenes and
triterpenes, which have been associated with antimicro-
bial, antinflammatory, antioxidant and gastroprotector
properties (Heral et  al., 1998; Baggio et  al., 2003; Abad
et al., 2006; Morales et al., 2008). EOs obtained from B.
notosergila Griseb. (Cobos et al., 2001), B. latifolia Pers.
and B. prunifolia Kunth (Rojas et al., 2007), B. elaeoides
J. Rémy and B. magellanica Pers. (Simonsen et  al.,
2009) showed antimicrobial properties. B. salicifolia
(Ruiz & Pav.) Pers. produces volatile compounds with
insect repellent properties (Garcia et al., 2005). The EO
obtained from B. dracunculifolia DC. possesses antiul-
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cerogenic properties associated with a high content of
E-nerolidol (Kloppel et  al., 2007). Additionally, caffeic
acid and pectolinaringenin isolated from the ethanol
extract of B. uncinella showed anti-leishmanial effects
(Passero et al., 2011).
Baccharis uncinella is used by the Laklaño Indians
(Santa Catarina, Brazil) as a sedative as well as to “regu-
late blood pressure”. The Shokleng Indians, who now
designate themselves as Laklaño, are recognized as a
group since the XVIII century. Living in the valleys and
plateau borders in the South of Brazil since 1914, the con-
For personal use only.
tacted group (approximately 800 individuals) was more
intensively exposed to the dominant society at the State
of Santa Catarina (Ibirama Indian Reserve). The inter-
cultural exchange introduced the use of salt and alcohol
beverages (Urban, 1985), as well as diseases unknown
to the Indians; moreover, medicinal plants from several
sources and cultural backgrounds were introduced by
local authorities and health providers (Santos, 1973).
Mr. Congo Patté, recognized up to this day as a medici-
nal plant expert at the Laklaño Reserve, was yet to be born
when a 1932 census listed 106 Laklaño individuals living
in the area (Henry, 1941). He speaks at easy of “heaviness
in the head”, nape pain, nausea and discouragement as
the symptoms indicative of high blood pressure; more-
over, he is ready to recognize the “cooking salt” as danger-
ous to older people’s health and recommends the use of
B. uncinella (known as vassourinha) tea to be used daily
for various days. He remembers that during the tense
period marked by land dispute with loggers, he used to
recommend that men avoid cachaça (Brazilian distilled
alcohol from sugar cane) and frequently used the same
B. uncinella tea in order to calm down.
CNS relevant components were identified in EOs
obtained from B. uncinella leaves collected in the South
of Brazil plateau above 1000 m of altitude (Ascari et al.,
2009). Noteworthy, significant differences in the EOs
composition were observed with different collections:
while the sample from the Paraná State (altitude 1050
m) presents low levels of monoterpenes (~2%) and a
high proportion of caryophyllene oxide (~16%) and
α-eudesmol (7.5%), the sample from Santa Catarina
 Pharmaceutical Biology
State (locality of Vacas Gordas, altitude 1360 m) contains
over 36% of monoterpenes (12.9% α-pinene and 9.9%
limonene) but a low proportion of caryophyllene oxide
(2.9%) and α-eudesmol (1.9%) (Ascari et al., 2009).
Given the traditional uses and relevance attributed
by the Laklaños to B. uncinella, the aim of this study was
to compare EOs obtained from two different areas of
Southern Brazil in terms of EO composition and sedative
effects.
Materials and methods
Ethnopharmacological survey
Ethnopharmacological data obtained for “vassourinha”,
Baccharis uncinella are part of a broader study car-
ried out at the Laklaño Indians land reserves known as
Terra Indígena Ibirama (State of Santa Catarina, Brazil),
between August 1999 and September 2001. The survey
was mainly conducted with three elderly native plant spe-
cialists, Mr. Congó Patté, and the ladies Iocô Uvanhecu
and Ngãvene Patté. “Vassourinha” tea is prepared with
a handful of leaves in a litter of cold water to be slowly
heated until boiling (decoction). The tea is to be drunk
for various days, by adults and the elderly, with the pur-
pose of regulating blood pressure and as a calming tea.
Plant material
The leaves of B. uncinella were initially collected in the
district of Ponta Grossa, State of Paraná, Brazil (sample
BU-PR, altitude of 1050 m, 25°06′23′′ South and 50°00′39′′
West) in July of 2005. A second collection was done close
to the Laklaño Reserve, at the locality of Pouso Redondo,
State of Santa Catarina, Brazil (sample BU-SC, alti-
tude of 440 m, 27°16′19′′ South and 49°49′60′′ West) in
July of 2006. Voucher specimens were deposited at the
Herbarium of the Universidade Estadual de Ponta Grossa
(HUPG-13105). Both collections were treated as follows:
plant material was dried for 7 days at room temperature,
the leaves separated and conserved at −18°C until the oil
extraction procedure.
Steam distillation and analyses of the EOs
The EO samples were obtained by hydrodistillation
with the use of aluminum and glass equipment. Close
to 1.2 kg of each milled leaves sample was submitted to
4 h distillation, and the EO collected in ethyl ether. The
organic solutions were dried with anhydrous Na2SO4,
with repeated filtrations and evaporations to yield 9.60 g
of BU-PR and 10.32 g BU-SC.
The oil samples were analyzed in a Varian® CP-3800
Gas Chromatograph coupled to a Saturn® 2000 Mass
Spectrometer using the software Saturn® GC-MS
Workstation 5.51, operating in the EI mode at 70 eV with a
mass range of 40–650 m/z and at a sample rate of 1.0 scan
s−1. An apolar capillary column CP-Sil-8 CB Low Bleed/
MS (30 × 0.25 mm i.d., 0.25 µm) film was used with the fol-
lowing conditions: split ratio of 1/50, 250°C for the injec-
tor and 240°C for the interface. The oven temperature was

programmed for 60°C for the first 3 min, raising at a rate
of 5°C/min to reach 220°C, remaining as such for slightly
over 15 min. The retention indices (RI) were calculated by
using the data from a series of n-alkane (C10-C26) injec-
tions in the same chromatographic conditions as those
used for the oil samples (Van den Dool & Kratz, 1963).
The components were firstly identified by comparing the
obtained mass spectra with those of the equipment data
bank, following by comparison of retention indices with
published data (Adams, 1995; Pherobase, 2010). Isolated
standards of heptanal, α-pinene, β-pinene, limonene,
linalool, caryophyllene, spathulenol, viridiflorol, gua-
iacol, camphor, trans-anetol, safrol, thymol, eugenol
and tert-butil-hydroxytoluene were used to validate the
system and guarantee the reliability of the calculated
indices.
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The relative composition of each oil sample was deter-
mined by using a Shimadzu Gas Chromatograph 14 B
with a Flame Ionization Detector (GC-FID), an OV-5 col-
umn (30 m × 0.25 mm d.i. × 0.25 µm) under the following
conditions: N2 used as the carrier gas at a constant pres-
sure 80 kPa, a split ratio of 1/150, and an injection volume
of 1 µL of the oil diluted in ethyl ether, with detector at
300°C, and injector at 250°C. The initial column tempera-
ture was set to 50°C for 3 min, programmed for heating up
at a rate of 5°C/min to reach final temperature of 270°C,
completing with an isotherm of 8 min.
For personal use only.
Pharmacological assessment
Animals
Experiments were performed with 2 month old (35–45 g)
male (CF1) albino mice, purchased from Fundação
Estadual de Produção e Pesquisa em Saúde (FEPPS).
Animals were maintained in our own animal facility,
under controlled environmental conditions (22 ± 1°C,
12 h light/dark cycle), with free access to food [Nuvilab
CR1] and water, for at least 2 weeks before the experi-
ments. All procedures were carried out in accordance
with institutional policies on experimental animals han-
dling, which follows de NIH guidelines (NIH Guide for
Care and Use of Laboratory Animals, NIH publication
No. 85–23, 1985).
Drugs
Diazepam and pentobarbital were acquired from Sigma.
B. uncinella EOs (BU-PR and BU-SC) were diluted in 1%
Tween 80. Drugs and vehicles were administered intrap-
eritoneally, always as 10 mL/kg of body weight.
Hypnotic activity
Mice (n = 8–10) were treated i.p. with saline, 1% Tween 80,
diazepam (2 mg/kg, positive control), or 50 or 100 mg/kg
of BU-PR or BU-SC. Thirty minutes later mice received
sodium pentobarbital (35 mg/kg, i.p.). The sleeping
time (time elapsed between loss and recovery of right-
ing reflex) was recorded; a cut-off time of 180 min was
adopted. Results were analyzed by means of ANOVA/
SNK (Linck et al., 2009).
© 2012 Informa Healthcare USA, Inc.
Sedative effects of Baccharis uncinella essential oils  115
Hypothermic effects
Groups of mice (n = 6) were treated i.p. with 50 or
100 mg/kg of BU-PR, BU-SC or controls (saline and
1% Tween 80). The body temperature measured with a
sensor probe of a digital thermometer (inserted at 1 cm
into the rectum) was recorded before treatments (time
0) and 15, 30, 60 and 120 min after drug administration.
Pentobarbital (50 mg/kg, i.p.) was used as the reference
drug. Results were analyzed by repeated measures
ANOVA followed by SNK (Dallmeier & Carlini, 1981).
Locomotion
The method was adapted from Linck et al. (2009). Activity
cages (45 × 25 × 20 cm, Albarsch Electronic Equipments),
equipped with three parallel photocells, automatically
record the number of crossings. Animals (n = 8–10) were
individually habituated to an activity cage for 10 min
before receiving the following treatments (i.p.): saline,
1% Tween 80, BU-PR and BU-SC 50 or 100 mg/kg. The
animals returned to the activity cages 30 min after treat-
ments and the number of crossings were recorded for
15 min. Diazepam 1 mg/kg was used as positive control.
Results were analyzed by means of ANOVA/SNK.
Results
Chemical composition of the EOs
Table 1 resumes the chemical analysis of BU-PR (over
1000 m altitude) and BU-SC (approximately 400 m alti-
tude). As can be seen, BU-PR presents a lower monot-
erpene content; the only monoterpene identified was
the oxygenated terpinen-4-ol (0.31%), resulting in a low
monoterpene/sesquitepene ratio (0.004). The main com-
ponents identified in this oil sample were sesquiterpenes:
spathulenol (32.93%), caryophyllene oxide (27.78%),
viridiflorol (5.29%) and α-cadinol (2.42%). These oxy-
genated compounds respond, therefore, for the majority
(68.42%) of the sample.
In a clearly distinct manner, the BU-SC sample presents
a sizable monoterpene content as well as non-oxygenated
sesquiterpenes, resulting in a monoterpene/sesquither-
pene ratio of 0.31. Only non-oxygenated monoterpenes
were identified (20.22%), of which α-pinene (6.42%) and
limonene (7.21%) are the major components. The non-
oxygenated sesquiterpenes identified comprise 38.63%
of the sample, with caryophyllene (26.13%) as the most
abundant component. The two oxygenated sesquit-
erpenes identified in BU-SC are spathulenol (13.39%)
and caryophyllene oxide (13.26%), which are often the
major components of EOs obtained from Baccharis spp.
(Agostini et al., 2005; Lago et al., 2008; Retta et al., 2009).
CNS activity of the EOs
A significant increase (F4.51 = 16.2; p < 0.01) in sleep-
ing time was induced by BU-PR at 100 mg/kg (but not
50 mg/kg); the effect of BU-PR 100 mg/kg was compa-
rable to diazepam. BU-SC likewise produced a significant
increase (F4.50 = 39.7; p < 0.01) in sleeping time, at both 50
 116  J. Ascari et al.
and 100 mg/kg; these doses had equal activity. At 100 mg/ BU-SC significantly and comparably decreased body tem-
kg, BU-SC was significantly (p < 0.05) more active than perature at 15 and 30 min post treatment, while at 60 min
BU-PR (Figure 1). post treatment only the hypothermic effect of pentobar-
Repeated measures ANOVA confirmed that the body bital was still at place. There was no significant difference
temperature was significantly decreased (F6.51 = 6.73; between the two doses of BU-PR and/or BU-SC.
p < 0.01) by all treatments in comparison to controls Figure 3 shows the results of treatments on locomo-
(Figure 2). The post hoc analyses showed that BU-PR and tion. A significant decrease (F6.56 = 17.5; p < 0.01) in loco-
motion was induced by BU-PR and BU-SC at both doses;
Table 1.  Relative compositions of volatile components of the
the differences between samples or doses of the same
two essential oils samples of Baccharis uncinella collected near sample did not reach statistical significance.
(BU-SC) or far (BU-PR) from the Laklaño reservation.
Component BU-SC % BU-PR % RI*
Discussion
α-thujene 2.91 932
α-pinene 6.42 940 In agreement with the claims from the Laklaño Indians,
β-pinene 3.68 980 we provide evidence that EOs obtained from Bacharis
limonene 7.21 1032 uncinella possess sedative properties given that dimin-
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terpinen-4-ol 0.31 1180 ished locomotion, hypothermia and hypnosis is a classic

α-copaene 2.93 0.27 1376 triad indicative of hypno-sedative effects. Differences in
β-cubebene 0.32 1391 the composition of the samples BU-PR and BU-SC were
β-elemene 0.22 1393 reflected in differences in hypnotic activities. Though
β-caryophyllene 26.13 0.92 1419 the effects of EOs in blood pressure was not studied,
aromadendrene 0.51 1440 considering the relationship between blood pressure
α-caryophyllene 3.63 1456 and stress, it is arguable that the perception of the useful-
γ-gurjunene 1.81 1472 ness of B. uncinella in regulating blood pressure can be
β-chamigrene 0.25 1476 in part associated with its sedative effects. Nonetheless,
germacrene D 1.88 t 1480 for a clear and comprehensive pharmacological profile
of these EOs further studies, including additional animal
For personal use only.

α-muurolene 1.34 1499

γ-cadinene 2.30 1510
β-cadinene 4.06 0.44 1522
trans-calamenene 0.56 1529
α-calacorene 2.23 1550
spathulenol 13.39 32.93 1578
caryophyllene oxide 13.26 27.78 1583
viridiflorol t 5.29 1590
α-cadinol t 2.42 1654
Sample yielding (w/w, %) 0.86 0.80
Total identified (%) 85.50 79.90
Monoterpenes (%) 20.22 0.31
Sesquiterpenes (%) 65.28 79.59 Figure 2.  Hypothermic effects of Baccharis uncinella essential
Monoterpene/Sesquiterpene 0.310 0.004 oils (BU-PR and BU-SC, 50 and 100 mg/kg i.p.). Pentobarbital
*RI: mean of the retention indices of each component. 50 mg/kg was used as positive control. * = p < 0.05; ** = p < 0.01 vs.
t: trace amount, below 0.10%. control. ANOVA/SNK.

Figure 1.  Effects of Baccharis uncinella essential oils (BU-PR and BU-SC, 50 and 100 mg/kg, i.p.) on pentobarbital-induced sleep in mice.
Diazepam 2 mg/kg was used as positive control. Each column represent the mean ± S.E.M. (n = 8–10). * = p < 0.05; ** = p < 0.01 vs. control
ANOVA/SNK.

 Pharmaceutical Biology

Figure 3.  Effects of Baccharis uncinella essential oils (BU-PR and BU-SC, 50 and 100 mg/kg, i.p.) on spontaneous locomotor activity. Each
column represents the mean ± S.E.M. (n = 8–10). ** = p < 0.01 vs. control. ANOVA/SNK.
models and dose ranges, would be necessary. Though the
activity cannot be related to components of EOs, sedative
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effects were also seen with B. serraefolia DC. methanol
extract, which was effective against strychnine- and
cardiazol-induced seizures, and increased sleeping time
(Tortoriello & Santamaria, 1996).
The major components of the samples here studied
are monoterpenes and sesquiterpenes. As mentioned
earlier, among volatile terpenes of natural origin several
are known to be centrally active (Heuberger et al., 2010).
Some of these can be found in EOs from Bacharis spe-
cies, such as α-pinene, limonene, linalool, myrcene,
α-terpineol, and the sesquiterpenes eudesmene, α- and
For personal use only.
β-eudesmol, and caryophyllene oxide (Agostini et  al.,
2005; Lago et  al., 2008; Ascari et  al., 2009; Retta et  al.,
2009; Simonsen et  al., 2009). Monoterpenes identified
in the EO samples here evaluated that can be related to
hypno-sedative properties include α-pinene (Sayyah
et al., 2004) and limonene (Vale et al., 1999; 2002).
In contrast, only a few volatile sesquiterpenes present-
ing CNS activity are currently known. The sesquiterpenes
caryophyllene oxide and β-selinene (= β-eudesmene)
isolated from the hexane extract from Psidium guayava
var. minor (Myrtaceae) Mattos leaves potentiated pen-
tobarbital-induced sleep and increased the latency for
PTZ-induced convulsions in mice; additionally, blockade
of extracellular Ca2+ was observed in isolated guinea-pig
ileum with the hexane extract and its fractions containing
both sesquiterpenes (Meckes et  al., 1997). β-Eudesmol
was found to be one of the volatile active principles of
the Chinese medicinal herb Atractylodes lancea DC.
(Asteraceae) with antagonist properties useful against
organophosphorous anticholinesterase agents intoxica-
tion (Chiou et  al., 1997). Experimental data show that
β-eudesmol prevents convulsions and lethality induced
by electroshock, but not those induced by PTZ or picro-
toxin (Chiou et  al., 1995). With a very similar chemical
structure, α-eudesmol protects the development of
post-ischemic brain injury in rats by blocking ω-Aga-
IVA-sensitive Ca2+ channels (Asakura et  al., 2000). The
structure resemblance of β-eudesmol and β-selinene is
likely to indicate relevant characteristic patterns com-
mon to these two compounds that are relevant for central
nervous system activity.
© 2012 Informa Healthcare USA, Inc.
Sedative effects of Baccharis uncinella essential oils  117
It has been documented that EOs from aerial parts
of B. uncinella obtained from Santa Catarina highlands
(Ascari et al., 2009) and the state of Rio Grande do Sul,
Brazil (Agostini et  al., 2005; Frizzo et  al., 2001, 2008)
contain 3–60% of monoterpenes. The volatile metabo-
lites in Baccharis are likely to be influenced by ecologi-
cal and geographical factors as luminosity (Silva et al.,
2006), rainfall regime (Ferracini et  al., 1995), ground
mineral content (Silva et  al., 2007) and/or interaction
with insects and predators (Damasceno et  al., 2010),
and the occurrence of chemotypes based on sesqui-
terpenes has been also reported (Frizzo et  al., 2008).
It is therefore arguable that the distinct composition of
BU-SC and BU-PR are related to ecological or genetics
factors; it is to be expected that the differences observed
in these samples CNS activities are consequent to these
differences in composition. The sedative effect is likely
to be related to sesquiterpenes (e.g., carophyllene oxide)
present in both samples, while the more marked hyp-
notic activity of BU-SC can be attributed to its higher
monoterpene content (e.g., α-pinene, limonene,
α-thujene). Given that the traditional preparation may
include compounds other than those found in the EO,
the contribution from other compounds to the phar-
macological properties alleged by Mr. Patté cannot be
excluded before different extracts from B. uncinella are
properly evaluated.
Conclusions
Clear differences exist in the phytochemical profile of
EOs obtained from the two collections, consistent with
the extensively documented effects of geographical/
ecological circumstances in EOs composition. The B.
uncinella EO sample collected closer to the Laklaño
Indians possesses a composition that better justifies
the claimed sedative properties. The differences in
composition likewise may be associated with the lack
of use of the same species in the State of Paraná. It is
arguable that this is yet another study that confirms
the value of traditional information to guide bioactiv-
ity assessment in medicinal plants. Moreover, the data
give notice to the ecological factors that can interfere
with such assessments if plant samples are collected
 118  J. Ascari et al.
without considering geographical factors that can ulti-
mately result in failure to correlate experimental data
with traditional claims.
Declaration of interest
Authors are grateful to CAPES for MSc fellowship (J A),
and CNPq for fellowships (EE and VML). Financial sup-
port was received from CNPq (478229/2006-2) and the
Fundação Araucária (047/2007-1573).
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