José Realino de Paula

Antifungal activity of volatile constituents of Eugenia dysenterica leaf oil

The essential oil from the hydrodistillation of Eugenia dysenterica leaves consisted mainly of beta-caryophyllene and alpha-humulene as the major sesquiterpene, while limonene and alpha-thujene were the major monoterpene hydrocarbons. The main oxygenated mono and sesquiterpene constituents were alpha-terpineol and beta-caryophyllene oxide, respectively. The oil was investigated against eight strains of Candida albicans, 35 strains of Cryptococcus neoformans var. neoformans, and two C. neoformans var. gattii isolated from HIV-infected individuals with candidosis or cryptococcal meningitis using the agar dilution method. Based on the minimal inhibitory concentration (MIC) values, the most significant results were obtained against Cryptococcus strains. It was observed that 22 strains were inhibited at a concentration of 250 microg/ml, whereas four exhibited potent inhibition with MIC values below 125 microg/ml against 10(6) UFC/ml organisms. We found MICs > or = 3.12 microg/ml for 91.6, 50 and 30% of all Cryptococcus strains in relation of amphotericin B, fluconazole and itraconazole, respectively.

Antifungal activity from Ocimum gratissimum L. towards Cryptococcus neoformans

Cryptococcal infection had an increased incidence in last years due to the explosion of acquired immune deficiency syndrome epidemic and by using new and effective immunosuppressive agents. The currently antifungal therapies used such as amphotericin B, fluconazole, and itraconazole have certain limitations due to side effects and emergence of resistant strains. So, a permanent search to find new drugs for cryptococcosis treatment is essential. Ocimum gratissimum, plant known as alfavaca (Labiatae family), has been reported earlier with in vitro activity against some bacteria and dermatophytes. In our work, we study the in vitro activity of the ethanolic crude extract, ethyl acetate, hexane, and chloroformic fractions, essential oil, and eugenol of O. gratissimum using an agar dilution susceptibility method towards 25 isolates of Cryptococcus neoformans. All the extracts of O. gratissimum studied showed activity in vitro towards C. neoformans. Based on the minimal inhibitory concentration values the most significant results were obtained with chloroformic fraction and eugenol. It was observed that chloroformic fraction inhibited 23 isolates (92%) of C. neoformans at a concentration of 62.5 microg/ml and eugenol inhibited 4 isolates (16%) at a concentration of 0.9 microg/ml. This screening may be the basis for the study of O. gratissimum as a possible antifungal agent.

Chemical variability in the essential oil of Hyptis suaveolens

The essential oils of Hyptis suaveolens plants collected from 11 localities of the Brazilian Cerrado region were investigated by GC-MS. Sabinene, limonene, biclyclogermacrene, beta-phellandrene and 1,8-cineole were the principal constituents. The results were submitted to principal component and chemometric cluster analysis which allowed three groups of essential oils to be distinguished with respect to the content of p-mentha-2,4(8)-diene, limonene/beta-phellandrene/gamma-terpinene and germacrene D/bicyclogermacrene. In patterns of geographic variation in essential oil composition indicated that the sesquiterpenes are mainly produced in the samples grown at lower latitudes.

The anxiolytic-like effect of an essential oil derived from Spiranthera odoratissima A. St. Hil. leaves and its major component, β-caryophyllene, in male mice

Spiranthera odoratissima A. St. Hil. (manacá) is used in folk medicine to treat renal and hepatic diseases, stomachache, headaches and rheumatism. A central nervous system (CNS) depressant effect of the hexane fraction from the ethanolic extract of this plant has been described. β-caryophyllene, the main component of this essential oil, is a sesquiterpene compound with anti-inflammatory properties that has been found in essential oils derived from several medicinal plants. This work is aimed to evaluate the pharmacological activity of the essential oil obtained from S. odoratissima leaves (EO) and its major component on the murine CNS; we aimed to evaluate a possible anxiolytic-like effect and the underlying mechanisms involved. In an open field test, EO (500 mg/kg) and β-caryophyllene (50, 100 and 200 mg/kg) increased the crossing frequency (P<0.05) and, EO (250 and 500 mg/kg) and β-caryophyllene (200 mg/kg) increased the time spent in the center (P<0.05) without altering total crossings of the open field. EO and β-caryophyllene did not alter the number of falls in the rota-rod test (P>0.05). In the pentobarbital-induced sleep test, EO (500 mg/kg) and β-caryophyllene (200 and 400 mg/kg) decreased the latency to sleep (P<0.05), and EO (125, 250 and 500 mg/kg) (P<0.001) and β-caryophyllene (200 and 400 mg/kg) (P<0.05 and P<0.001) increased the sleep time. In anxiety tests, EO (500 mg/kg) and β-caryophyllene (100 and 200 mg/kg) increased head-dipping behavior (P<0.05) in the hole-board test, entries (P<0.05) into and time spent (P<0.05) on the open arms of the elevated plus maze (EPM), and number of transitions (P<0.05) and time spent in the light compartment (P<0.05) of a light-dark box (LDB). We further investigated the mechanism of action underlying the anxiolytic-like effect of EO and β-caryophyllene by pre-treating animals with antagonists of benzodiazepine (flumazenil) and 5-HT(1A) (NAN-190) receptors prior to evaluation using EPM and LDB. The anxiolytic-like effects of EO were significantly reduced by pre-treatment with NAN-190 (P<0.05) but not flumazenil (P>0.05). The anxiolytic-like effects of β-caryophyllene were not blocked by either NAN-190 or flumazenil (P>0.05). In conclusion, these results suggest that the essential oil derived from S. odoratissima produces an anxiolytic-like effect without altering motor performance and that this effect is mediated by 5-HT(1A) but not via benzodiazepine receptors. In addition, the major component, β-caryophyllene, also has an anxiolytic-like effect that may contribute to the effects of EO, but this effect does not seem to be mediated via 5-HT(1A) or benzodiazepine receptors.

Wound Healing and Anti-Inflammatory Effect in Animal Models of Calendula officinalis L. Growing in Brazil.

Calendula officinalis is an annual herb from Mediterranean origin which is popularly used in wound healing and as an anti-inflammatory agent. In this study, the ethanolic extract, the dichloromethane, and hexanic fractions of the flowers from plants growing in Brazil were produced. The angiogenic activity of the extract and fractions was evaluated through the chorioallantoic membrane and cutaneous wounds in rat models. The healing activity of the extract was evaluated by the same cutaneous wounds model through macroscopic, morphometric, histopathologic, and immunohistochemical analysis. The antibacterial activity of the extract and fractions was also evaluated. This experimental study revealed that C. officinalis presented anti-inflammatory and antibacterial activities as well as angiogenic and fibroplastic properties acting in a positive way on the inflammatory and proliferative phases of the healing process.

Repellent activity of plant-derived compounds against Amblyomma cajennense (Acari: Ixodidae) nymphs

Repellence responses of Amblyomma cajennense nymphs to callicarpenal, intermedeol, Hyptis suaveolens essential oil, extract of Melia azedarach, Cymbopogon nardus, Spiranthera odoratissima, Chenopodium ambrosioides, Ageratum conyzoides, Mentha pulegium, Ruta graveolens, and Memora nodosa were studied. Among these the extract of C. nardus stood out because of the long-lasting repellence, maintaining, in the highest concentration, 35h of protection against 90% of the nymphs. The essential oil of H. suaveolens and the extracts of C. ambrosioides and A. conyzoides showed good repellence index (66%) when applied in high concentrations. However, greater protection could be obtained at higher concentrations but with a shorter repellence time. Callicarpenal, intermedeol, extract of M. Pulegium, and M. nodosa leaves showed moderate repellence in high concentrations. Extracts from M. azedarach, R. graveolens, S. odoratissima, and M. nodosa roots showed little or no repellent effect. These results show that some plant extracts may represent a promising alternative in the control of infestations by A. cajennense.

Antidepressant-like effect of Lafoensia pacari A. St.-Hil. ethanolic extract and fractions in mice.

ETHNOPHARMACOLOGICAL RELEVANCE Lafoensia pacari A. St.-Hil. (Lythraceae) has been referred in Brazilian traditional medicine for the treatment of different diseases, among them depression. Nevertheless, there are not studies about this possible effect on the central nervous system (CNS). AIM OF THE STUDY To evaluate the antidepressant-like effects of the ethanolic extract of Lafoensia pacari (PEtExt) and its fractions on the performance of male mice. MATERIALS AND METHODS Antidepressant activity was studied using forced swimming (FST) and tail suspension (TST) tests, and motor activity in the open-field test. The ethanolic extract of Lafoensia pacari (PEtExt) were administered acutely (1.0 g/kg, p.o.), for 21 days (100, 300 mg, and 1.0 g/(kg day), p.o.), three administration in a 24-h period (1.0 g/kg, p.o.), and the fractions for 21 days. Imipramine (15 mg/(kg day), p.o.) was used as the control positive. RESULTS The PEtExt significantly reduced immobility time in FST and TST, without affecting the motor activity. Only the chloroformic fraction (50 mg/(kg day), p.o.) increase the latency to immobility and decrease the immobility time in the FST. CONCLUSIONS These data indicate that the extract of Lafoensia pacari A. St.-Hil. possesses antidepressant-like properties in mice.

Estudo farmacognóstico das folhas de Pimenta pseudocaryophyllus (Gomes) L.R. Landrum - Myrtaceae

The goal of this research was to obtain quality control parameters for the identification of the plant drug of Pimenta pseudocaryophyllus (Gomes) L. R. Landrum that occurs in Brazilian savannah. So, two samples were collected, one in Minas Gerais state - Brazil, and the other one in Brasilia city - DF - Brazil, and the vouchers were deposited in the UFG Herbarium code number UFG-27159 and Ezechias Paulo Heringer Herbarium code number 21745-0, respectively. Part of the botanical material in both samples was prepared for microscopic analysis. The other one was air-dried, crushed to powder and used for phytochemical screening. The study has shown that P. pseudocaryophyllus leaves are hypostomatic, there are abundant unicellular trichomes in abaxial surface, there are secretory cavities with oily content and epidermal cells have thick wall. Tannins, phenolic compounds and flavonoids were detected in the phytochemical screening. The tannins had been quantified by different methods involving colorimetric and protein precipitation reactions, getting different results. This fact calls the attention for the necessity of specifying the method used together with the minimum tannins limit established. The results configure as important parameters for the quality control of the plant material and denote that the species in study take an important potential as herbal medicine that it deserves to be investigated in pharmacologic and toxicologic studies.

Ação do óleo essencial de Syzygium aromaticum (L.) Merr. & L.M.Perry sobre as hifas de alguns fungos fitopatogênicos

Action of Syzygium aromaticum (L.) Merr. & L.M.Perry essential oil on the hyphae of some phytopathogenic fungi. Currently, the use of alternative methods to control diseases and pests in agriculture has been a recognized and necessary practice to minimize damages to the environment and public health. This study aimed to investigate the action of clove (Syzygium aromaticum (L.) Merr. & L.M.Perry) essential oil on the in vitro mycelial growth of the phytopathogenic fungi Rhizoctonia solani, Fusarium solani, Fusarium oxysporum and Macrophomina phaseolina. Analysis by gas chromatography-mass spectrometry allowed the identification of eugenol (83.6%), eugenyl acetate (11.6%) and caryophyllene (4.2%). Microscopic evaluation of mycelia showed several morphological changes such as presence of vacuoles, cell content disorganization, decreased cell wall clearness, intense fragmentation and lower turgescence of hyphae. Clove essential oil showed fungicidal activity at 0.15% on the growth of R. solani, F. oxysporum and F. solani, but not for M. phaseolina. These results indicate favorable perspectives for future use of clove essential oil to control these phytopathogens in agriculture.

Investigation of Ehrlich ascites tumor cell death mechanisms induced by Synadenium umbellatum Pax.

ETHNOPHARMACOLOGICAL RELEVANCE Synadenium umbellatum Pax. is widely found in South America and empirically used in Brazil for the treatment of several diseases, mainly cancer. The aim of the study was to investigate cell death mechanisms induced by Synadenium umbellatum Pax. using Ehrlich ascites tumor (EAT) cells, as well as the myelotoxicity potential of this plant. MATERIALS AND METHODS S. umbellatum cytotoxicity was evaluated in EAT cells by trypan blue exclusion and MTT reduction test and the mechanisms involved in EAT cell death were investigated by light and fluorescence microscopy, flow cytometry and immunocytochemistry. Investigation of S. umbellatum myelotoxicity was performed by clonogenic assay of colony forming unit- granulocyte macrophage (CFU-GM). RESULTS AND CONCLUSION Our results demonstrated that S. umbellatum decreased the viability of EAT cells using both methods. Morphological analyses revealed that S. umbellatum-treatment induced EAT cell death by apoptotic pathway. We demonstrated the occurrence of reactive oxygen species (ROS) overgeneration, increased intracellular Ca(2+) concentration, alteration in mitochondrial membrane potential, phosphatydylserine externalization, and activation of caspases 3, 8, and 9. However, S. umbellatum produced myelotoxicity in bone marrow cells in a concentration-dependent manner. In comparison to EAT cells, the effects of S. umbellatum in bone marrow cells were 8-fold lower. Taken together, our results showed that S. umbellatum induced apoptosis in EAT cells at several levels and seems more toxic to tumor cells than to normal bone marrow cells.