Murilo Marchioro

Ocimum basilicum leaf essential oil and (-)-linalool reduce orofacial nociception in rodents: a behavioral and electrophysiological approach

The present study investigated the antinociceptive effects of Ocimum basilicum L. (Lamiaceae) leaf essential oil (LEO) and (-)-linalool (LIN) in formalin (2%)-, glutamate (25 µM)- and capsaicin (2.5 µg)- induced orofacial nociception models in mice. The involvement of these substances was further evaluated on the neuronal excitability of the hippocampal dentate gyrus. Male mice (n=8/group) were pretreated separately with LEO and by LIN (50, 100, and 200 mg/kg, i.p.), morphine (5 mg/kg, i.p.) and vehicle (saline + Tween 80 0.2%), before injection of nociceptive agent into the right upper lip (perinasal area). The LEO and LIN reduced the nociceptive face-rubbing behaviour in both phases on formalin test. LEO and LIN, at high doses, produced significantly antinociceptive effect in the capsaicin and glutamate tests. In hippocampal slices, LEO inhibited the population spike generated by stimulation of the hylus (antidromic stimulation), with an IC50 of 0.1±0.05 mg/mL. This response was reversibly blocked by lidocaine (0.5 mg/mL), a known voltage-dependent sodium channel antagonist and by LIN (0.5 mg/mL). Our results suggest that LEO and LIN modulate neurogenic and inflammatory pain in the tests of orofacial nociception induced by formalin, capsaicin and glutamate. Part of these effects may be associated with decreased peripheral and central neuronal excitability.

Postnatal neurogenesis in the medial cortex of the tropical lizard Tropidurus hispidus.

Young, adult and presumed old specimens of the tropical lizard Tropidurus hispidus, living in an almost steady warm habitat, have been the subjects of a 5-bromodeoxiuridine immunocytochemical study to label proliferating brain cells. All animals showed abundant 5-bromodeoxiuridine-labeled nuclei in the ependyma of their telencephalic lateral ventricles, with these being especially abundant in the medial cortex ependyma. Surprisingly, adult animals displayed higher numbers of labeled nuclei when compared with those of young specimens. In a second experiment, in order to check the evolution of ependymal-labeled nuclei, adult specimens were allowed 4 h or 2, 4, 7, 15 or 30 days of survival after the 5-bromodeoxiuridine pulse. Most labeled nuclei appeared isolated at short survival times (4 h and 2 days after the 5-bromodeoxiuridine pulse) but from day 4 and beyond, labeled nuclei appeared in couples or groups usually located in the ependyma. Labeled nuclei with vertical fusiform appearance in the inner plexiform layer or even recruited in the medial cortex cell layer were assumed to be migratory. These presumed migratory nuclei were unexpectedly few (less than 30%) when compared with other lizards, and they appeared much later; at 15 and 30 days after the pulse. This situation resembles that of mammals where only a small proportion of postnatally generated neurons can develop and survive.

Effects of the aqueous extract from Hyptis pectinata leaves on rodent central nervous system

The effects of the aqueous extract (AE) from Hyptis pectinata leaves was studied on rodent central nervous system (CNS). Pharmacological screening, open field, forced swimming, apomorphine-induced hypothermia, elevated plus maze and thiopental-induced sleep tests were used in male and female Swiss mice and Wistar rats. The AE was admnistered orally in single doses 30 minutes before each test. In the screening test the AE (400 mg/kg, p.o., n =3) decreased the frequency of grooming behavior. In the open field test it decreased the amount of time rats spent grooming (AE400: 51.3 ± 8.8, q = 5.513, P < 0.01, n = 10), similarly to diazepam (control: 107.3 ± 14.2; diazepam: 15.4 ± 4.3, q = 9.049, P < 0.001, n = 10). In the forced swimming test the AE (400 mg/kg) decreased the immobility time (con: 181.3 ± 7.2 s; imip: 91.6 ± 8.9 s, q = 7.958, P < 0.001; AE400: 111.6 ± 14.5 s, q = 6.193, P < 0.001, n = 9), indicating a possible antidepressive effect. This was confirmed in the apomorphine-induced hypothermia model where the extract (200 mg/kg) antagonized the apomorphine effect (AE200: -0.27 oC, q = 5.588, P < 0.001, n = 10), similarly to imipramine (imipramine: 5 mg/kg) (control: -1.08 oC; imipramine: 0.02 oC, q = 7.589, P < 0.001, n = 10). No effect of the AE was detected in the elevated plus maze and in the sleeping time induced by thiopental models. Our results suggest a possible antidepressive effect of the aqueous extract obtained from the leaves of Hyptis pectinata.

Epidemiological aspects of dengue in Aracaju, State of Sergipe, Brazil

INTRODUCTION The dengue fever remains to be a disease of serious public health concern, and its incidence has increased in the past decades. This study aimed to characterize the epidemiological incidence of dengue in the period 2001-2010. METHODS This is an epidemiological study of dengue in the municipality of Aracaju, state of Sergipe, in the period between 2001 and 2010, whose data were obtained from the Information System of Diseases Notifications. A descriptive analysis of the number of confirmed cases of dengue, according to year, semester, sanitary district, age, and sex, was performed. RESULTS There were 16,462 confirmed cases, especially in 2008, which obtained the highest incidence of the disease, with 10,485 confirmed cases. The first semester obtained the highest registration of cases during the years of research; this was predominated by females between 15 and 49 years old. With regard to the territorial distribution, the second district of the municipality obtained the highest number of cases. CONCLUSIONS In 2008, in the City of Aracaju, SE, a significant increase in the proportion of dengue cases compared with other years was verified. However, a fast decline in the other years was observed, possibly because of the intensification of preventive actions to combat the mosquito that transmits the dengue virus.

Benzodiazepine-Like Effects of the Alcohol Extract from Erythrina velutina. Leaves: Memory, Anxiety, and Epilepsy

Abstract In this work, we studied the effects of the alcohol extract (AE) from Erythrina velutina. Willd (Fabaceae) leaves in animal models of anxiety (elevated plus maze; EPM), memory (inhibitory avoidance test), and epilepsy (pentylenetetrazol-induced convulsions and pentylenetetrazol-induced kindling seizure). In the EPM test, at the dose of 20 mg/kg, the AE significantly increased the percentage of entries into the open arms (p = 0.03). In the inhibitory avoidance test, greater test than training latencies were observed for the control group but not for the AE (10 mg/kg) (p = 0.112) and diazepam (p = 0.331) treated groups during the acquisition phase of the test. During the consolidation and retrieval phases, there were differences between training and test latencies for all three groups. In the pentylenetetrazol-induced convulsion test, the AE (100 mg/kg) increased the latency to death when compared with the control group (p < 0.05) and reduced the kindling behavior induced by low doses of pentylenetetrazol. Together, the effects described for the AE of E. velutina. leaves on rodent central nervous system resemble the profile of effects of benzodiazepines and could be interpreted as an interaction of the extract with GABAergic circuitries.

Relaxant effect of the aqueous extract of Erythrina vellutina leaves on rat vas deferens

The effect of the Aqueous Extract from the leaves of Erythrina vellutina (AE) on rat vas deferens preparation was evaluated in this work. The AE inhibited the muscle contractions induced by electrical field stimulation (EFS) in a concentration-dependent manner. This inhibition was not affected by atropine (10-5M), propanolol (10-5M), prazosin (10-5M) or yohimbine (10-5M), suggesting that there is no direct interaction of the AE with cholinergic nor adrenergic receptors. Incubation of vas deferens with the K+ channel antagonists, tetraethylamonium (10-6M) or 4-aminopyridine (10-6M) had also no effect on the AE-induced inhibition. On the other hand, glibenclamide (10-6) significantly attenuated the effect of the AE, suggesting a possible involvement of ATP-dependent K+ channels. The AE (0.15 mg/mL) did not alter the contractions induced by noradrenaline (10-5M), ATP (10-4M) nor KCl (80 mM), against an interaction of the extract with post-synaptic sites. The data presented suggests that the inhibition of the electrically driven muscle twitches by the AE could be due to a pre-synaptic interaction of the extract with ATP-dependent K+ channels from vas deferens sympathetic neurons.

Essential Oil of Ocimum basilicum L. and (−)-Linalool Blocks the Excitability of Rat Sciatic Nerve

The racemate linalool and its levogyrus enantiomer [(−)-LIN] are present in many essential oils and possess several pharmacological activities, such as antinociceptive and anti-inflammatory. In this work, the effects of essential oil obtained from the cultivation of the Ocimum basilicum L. (EOOb) derived from Germplasm Bank rich in (−)-LIN content in the excitability of peripheral nervous system were studied. We used rat sciatic nerve to investigate the EOOb and (−)-LIN effects on neuron excitability and the extracellular recording technique was used to register the compound action potential (CAP). EOOb and (−)-LIN blocked the CAP in a concentration-dependent way and these effects were reversible after washout. EOOb blocked positive amplitude of 1st and 2nd CAP components with IC50 of 0.38 ± 0.2 and 0.17 ± 0.0 mg/mL, respectively. For (−)-LIN, these values were 0.23 ± 0.0 and 0.13 ± 0.0 mg/mL. Both components reduced the conduction velocity of CAP and the 2nd component seems to be more affected than the 1st component. In conclusion EOOb and (−)-LIN inhibited the excitability of peripheral nervous system in a similar way and potency, revealing that the effects of EOOb on excitability are due to the presence of (−)-LIN in the essential oil.

Structural organization of the cerebral cortex of the neotropical lizard Tropidurus hispidus

Lizards belonging to the Tropiduridae family are “sit-and-wait” foragers, relying mainly on visual identification to catch prey that cross their visual fields. Little is known about the neurobiology of Tropiduridae lizards. We have used neurohistological techniques to study the structural organization of the telencephalon of the neotropical lizard Tropidurus hispidus, paying special attention to the cerebral cortex. As revealed by the Nissl technique and Golgi staining, the telencephalon of T. hispidus follows the squamate pattern, with some differences: the lateral cortex appears relatively atrophic, and most of the neuronal somata of the dorsal cortex are dispersed without forming a conspicuous cell layer. Golgi staining has revealed ten different neuronal types in the three cortical layers, based on somata shape and dendritic morphology: the granular (unipolar, bipolar, and multipolar), pyramidal (normal, inverted, open, bipyramidal, and horizontal), spherical horizontal, and fusiform neuronal types. The axon direction could be traced in five of the subtypes. We have also studied the distribution of zinc-enriched terminals in the telencephalon of T. hispidus by the Neo-Timm method. Some portions of the cortex, septum, striatum, and amygdaloid complex stain heavily, with patterns resembling those described for other lizard families. Thus, T. hispidus appears to be an interesting representative of the Tropiduridae family for further neurobiological comparative studies.

Carvacrol prevents impairments in motor and neurochemical parameters in a model of progressive parkinsonism induced by reserpine

Parkinsons disease (PD) is a neurodegenerative disease characterized by a progressive degeneration of dopaminergic neurons in the substantia nigra pars compact (SNpc), with consequent depletion of dopamine in the striatum, which gives rise to the characteristic motor symptoms of PD. Although its etiology is unknown, several studies have suggested that oxidative stress plays a critical function in the pathophysiology of PD, and antioxidant agents could be helpful to slown down the dopaminergic neurodegeneration. Carvacrol (CA) is a phenolic monoterpene found in essential oils of many aromatic plants that presents antioxidant and neuroprotective effects. This study aimed to assess the effect of CA in a reserpine (RES)-induced rat model of PD. Male Wistar rats received 15 s.c. injections of 0.1 mg/kg RES or vehicle, every other day, concomitantly to daily i.p. injections of CA (12.5 or 25 mg/kg) or vehicle. Across the treatment, the animals were submitted to behavioral evaluation in the catalepsy test (performed daily), open field test (7th day) and assessment of vacuous chewing movements (12th, 20th and 30th days). Upon completion of behavioral tests, rats were perfused and their brains underwent tyrosine hydroxylase (TH) immunohistochemical analysis. Our results showed that CA (12.5 e 25 mg/kg) prevented the increase in catalepsy behavior and number of vacuous chewing movements, but failed to revert the decreased open-field locomotor activity induced by RES. In addition, CA in both doses prevented the decrease in TH immunostaining induced by RES in the SNpc and dorsal striatum. Taken together, our results suggest that CA shows a protective effect in a rat model of PD, preventing motor and neurochemical impairments induced by RES. Thus, the use of CA as a promising new strategy for the prevention and/or treatment of PD may be considered.

Characterization of NADPH Diaphorase- and Doublecortin-Positive Neurons in the Lizard Hippocampal Formation.

The lizard cortex has remarkable similarities with the mammalian hippocampus. Both regions process memories, have similar cytoarchitectural properties, and are important neurogenic foci in adults. Lizards show striking levels of widespread neurogenesis in adulthood and can regenerate entire cortical areas after injury. Nitric oxide (NO) is an important regulatory factor of mammalian neurogenesis and hippocampal function. However, little is known about its role in nonmammalian neurogenesis. Here, we analyzed the distribution, morphology, and dendritic complexity (Neurolucida reconstructions) of NO-producing neurons through NADPH diaphorase (NADPHd) activity, and how they compare with the distribution of doublecortin-positive (DCX+) neurons in the hippocampal formation of the neotropical lizard Tropidurus hispidus. NADPHd-positive (NADPHd+) neurons in the dorsomedial cortex (DMC; putatively homologous to mammalian CA3) were more numerous and complex than the ones in the medial cortex (MC; putatively homologous to the dentate gyrus). We found that NADPHd+ DMC neurons send long projections into the MC. Interestingly, in the MC, NADPHd+ neurons existed in 2 patterns: small somata with low intensity of staining in the outer layer and large somata with high intensity of staining in the deep layer, a pattern similar to the mammalian cortex. Additionally, NADPHd+ neurons were absent in the granular cell layer of the MC. In contrast, DCX+ neurons were scarce in the DMC but highly numerous in the MC, particularly in the granular cell layer. We hypothesize that NO-producing neurons in the DMC provide important input to proliferating/migrating neurons in the highly neurogenic MC.