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This study was aimed to microencapsulate fish oil (FO) in two biocompatible polymeric blends: gum arabic (GA)-maltodextrin (MD) and casein-pectin (CP)-MD. GA-MD microparticles and CP-MD microparticles were produced by spray-drying and complex coacervation and spray-drying, respectively. Encapsulation efficiency, particle size, moisture content, oxidative stability, and morphological properties were analysed. Encapsulation efficiencies of 51.2-56.8% (w/v) for GA and 64.7-67.9% (w/v) for CP preparations were found. GA particle sizes varied from 2 to 100 μm and from 2 to 120 μm for CP microparticles. Spherical forms with depressions in the topography of both systems were evidenced by scanning electron microscopy. Confocal microscopy evidenced surface oil on GA microparticles, corroborating encapsulation efficiency. CP was more efficient than GA to reduce oxidation, with maximum peroxide values (PVs) of 17.40 mmol/kg oil after 28 d at 40 °C/75% relative humidity (RH). Thus, CP is a promising biopolymeric blend for encapsulation of FO that provides protection against lipid oxidation.

Crude non-volatile oleoresin from Burseraceae species (breu), a non-timber forest product of the Amazon, still lacks methods for its quality control even though its pharmacological activities have been associated to the major triterpenes α-amyrin (αAA), β-amyrin, α-amyrenone, β-amyrenone and lupeol (LU). Its chemical characterization has usually been determined by gas chromatography (GC) using columns with phases of low polarity, which are ineffective in the separation of αAA and LU. The present study aimed to develop a GC method to characterize and quantify triterpenes in commercial breu with good selectivity and resolution. Columns with phases of different polarity (DB-35, DB-1701, DB-17HT and DB-Innowax) were tested, and DB-17HT separated αAA and LU without signal suppression or high retention of other constituents. This column allowed the quantification of breu triterpenes using flame ionization detection and cholesterol as a reference standard. Good similarity between the calibration curves of cholesterol and αAA was verified. Internal and external standardizations led to different (p < 0.05) quantifications, but the latter showed less deviation (RSD ≤ 3%). This work provides a reproducible GC method for the quality control of commercial breu, clearly evidencing the presence of lupeol and allowing the accurate quantification of major triterpenes. Resumo A oleorresina bruta não volátil das espécies de Burseraceae (breu), um produto florestal não madeireiro da Amazônia, ainda carece de métodos para seu controle de qualidade, embora suas atividades farmacológicas tenham sido associadas aos principais triterpenos α-amirina (αAA), β-amirina, α-amirenona, β-amirenona e lupeol (LU). Sua caracterização química tem sido geralmente determinada por cromatografia gasosa (CG), utilizando colunas com fases de baixa polaridade, que são ineficazes na separação de αAA e LU. O presente estudo teve como objetivo desenvolver um método de CG para caracterizar e quantificar triterpenos em breu comercial com boa seletividade e resolução. Colunas com fases de diferentes polaridades (DB-35, DB-1701, DB-17HT e DB-Innowax) foram testadas, e DB-17HT separou αAA e LU sem supressão de sinal ou alta retenção de outros constituintes. Esta coluna permitiu a quantificação de triterpenos do breu utilizando detecção de ionização de chama e colesterol como padrão de referência. A boa similaridade entre as curvas de calibração do colesterol e αAA foi verificada. Padronizações interna e externa levaram a quantificações diferentes (p < 0,05), mas a última apresentou menor desvio (RSD ≤ 3%). Este trabalho fornece um método de CG reprodutível para o controle de qualidade do breu comercial, evidenciando claramente a presença do lupeol e permitindo a quantificação precisa dos principais triterpenos.

Ethnopharmacological relevance Leishmaniasis is a neglected disease that affects millions of people around the world. Parasite resistance and the toxicity to the current treatments lead to the search for new effective molecules. Plants are widely used in traditional and indigenous medicine to treat different diseases. The oleoresin of the genus Protium, which is rich in volatile compounds active against different microorganisms, is among these plants. Aim The aim of this study was to evaluate the leishmanicidal potential of Protium altsonii (PaEO) and P. hebetatum (PhEO) (Burseraceae) oleoresins, as well as of three representative monoterpenes in their constitution: α-pinene, p-cymene and 1,8-cineole. Materials and methods Protium altsonii (PaEO) and P. hebetatum (PhEO) oleoresins and three of their constituents were tested in vitro on promastigotes and amastigotes-infected macrophages in different concentrations. Their toxicity for macrophages was analyzed by XTT assay and phagocytic ability. It was evaluated the ability of the compounds to induce NO production on treated-macrophages using Griess reaction and the effect of them in lipid profile on treated-parasite through Thin Layer Chromatography. Results: Our data showed that both essential oils have toxic effect on promastigotes and amastigotes of L. amazonensis in vitro in a dose-dependent manner. PaEO IC50 were 14.8 μg/mL and 7.8 μg/mL and PhEO IC50s were 0.46 μg/mL and 30.5 μg/m for promastigotes and amastigotes, respectively. Toxicity to macrophages was not observed at 50 μg/mL with both EOs. The compounds 1,8- cineole, α-pinene, and p-cymene inhibited amastigotes survival in a dose-dependent manner with IC50s of 48.4 μg/mL, 37 μg/mL, 46 μg/mL, respectively. Macrophage viability was around 90% even at 200 μg/mL and the phagocytic capacity was not altered in the treated-macrophages to up 50 μg/mL. The compounds were not able to modulate the nitric oxide production either at rest or LPS-activated macrophages. In addition, treated promastigote revealed an important change in their lipid profile after 48 h at 50 μg/mL in the presence of the compounds. Conclusions The results indicate that oleoresins of Protium genus are potent against Leishmania and α-pinene, p-cymene and 1,8-cineole have anti-Leishmania properties that could be explored in synergistic assays in order to develop new drug candidates.

Leishmaniases is a tropical disease caused by protozoa of the genusLeishmaniafor which the current treatment is expensive, besides increasing reports of parasite resistance. This study investigated the anti-Leishmania amazonensisactivity of the essential oil fromAloysia gratissima(AgEO) and guaiol, the major sesquiterpene constituent in the oil. Our results showed that AgEO killed promastigotes and intracellular amastigotes at an IC50of 25 and 0·16µg mL−1, respectively, while guaiol killed amastigotes at an IC50of 0·01µg mL−1. Both AgEO and guaiol were safe for macrophages up to 100µg mL−1, as evaluated by the dehydrogenase activity, membrane integrity and phagocytic capacity. AgEO and guaiol did not induce nitrite oxide (NO) in resting macrophages and inhibited the production of NO in lipopolysaccharide-stimulated macrophages. The ultrastructural analysis suggested that AgEO and guaiol act directly on parasites, affecting promastigotes kinetoplast, mitochondrial matrix and plasma membrane. Together, these results pointed out that AgEO and guaiol could be promising candidates to develop anti-Leishmaniadrugs.

Context 5-Aminolevulinic acid (5-ALA) is a prodrug used in photodynamic therapy (PDT) of tumors, including cancer of the oral mucosa. 5-ALA poorly penetrates oral tissues due to its high hydrophilicity, which impairs its local effects in PDT. Objectives To examine whether α-bisabolol (α-Bis) influences the 5-ALA permeability in the porcine buccal mucosa, to an extent that improves its application in PDT (which requires low permeation and high retention in the buccal mucosa). Methods In vitro permeability studies with 5-ALA (1% and 10% w/w) associated with α-Bis (1% to 20% w/w) in propylene glycol were carried out at 4 h and 24 h using porcine buccal mucosa in a modified Franz cell system. The in vitro release profiles (0.5 to 48 h) of the selected formulation and its respective control were determined using artificial membranes. Samples of buccal mucosa treated with the formulation were submitted to histopathological analysis, using a routine optical microscopy technique. Results The association of 1% 5-ALA and 5% α-Bis provided the best results; after 4 h of treatment with this formulation, the 5-ALA permeation was low and its retention in the mucosa was six-fold higher than that promoted by the control formulation (5-ALA alone). Histological analysis of the porcine buccal mucosa evidenced that 5% α-Bis altered the tissue morphology, which probably promoted 5-ALA retention. We concluded that 5% α-Bis is a potential adjuvant in formulations containing 5-ALA that could improve its retention after topical oral administration for the PDT treatment of cancer.

The leaf essential oils from Eugenia luschnathiana Klotzsch ex B.D. Jacks. and Myrciaria tenella (DC.) O. Berg growing in two different locations in southeastern Brazil were obtained by hydrodistillation and analyzed by GC and GC-MS. The oil profiles of individual accesses of E. luschnathiana from each location were characterized by the virtual absence of monoterpenes. The sesquiterpenes were of the caryophyllane (32%), guaiane (27%) and cadinane (19%) types in the Sao Paulo State access and caryophyllane (43%) and cadinane (28%) types in the Rio de Janeiro access. This is the first report on the composition of E. luschnathiana oil. Monoterpenes were predominant in both accesses of M. tenella (57-65%), and sesquiterpenes were mostly represented by the guaiane-eudesmane (16%) or caryophyllene-guaiane types (17%), respectively, in the Sao Paulo and Rio de Janeiro accesses. In monoterpene composition, the M. tenella oils qualitatively and quantitatively resembled that from a previous access from sout...

The leaf essential oils of three Myrtaceae species: Melaleuca leucadendra, Lophostemon confertus and Ugni molinae, non-native to Brazil and growing in the Rio de Janeiro Botanical Garden, were obtained by hydrodistillation and analyzed by GC and GC-MS. Oil yields from fresh leaves were 0.76%, 0.08%, and 0.04%, respectively. Sixty-seven percent of M. leucadendra oil was made up of monoterpenes, wherein 1,8-cineole was largely predominant (49%), followed by α-terpineol (7.6%) and terpinen-4-ol (4.3%). The sesquiterpene profile was characterized by a significant amount of viridiflorol (29%). Oil from L. confertus was 42% monoterpenes; the most predominant constituents being α-pinene (20.8%) and α-thujene (7.1%). Total sesquiterpenes (55.4%) comprised spathulenol (28%), globulol (14%), and aromadendrene derivatives (8.5%) as the main species. U. Molinae oil was exclusively composed of sesquiterpenes, with a predominance of β-elemene (44%) followed by β-caryophyllene (7.1%), and bicyclogermacrene (6.7%); guaiol-type alcohols made up 30% of its profile. To the best of our knowledge, this is the first time U. Molinae leaf oil composition has been described. The composition of M. leucadendra oil corresponded to a chemotype described for a species growing in Cuba, but was different from that produced by M. leucadendra growing in Egypt, Tunisia, and elsewhere in Brazil. The profile of the L. confertus leaf oil showed high similarity with those described for Australian native specimens, in particular in relation to the relevant presence of α-pinene, aromadendrenes, and the guaiol-type alcohols.

Despite the many biological activities reported for essential oils, their anti-inflammatory ability is relatively underexplored considering the wide variation in plant sources and in their volatile composition. Oils from Syzygium cumini Skells (SC) and Psidium guajava L. (PG) (Myrtaceae) have been described as having diverse pharmacological activities.The current study seeks to evaluate the anti-inflammatory activity of the essential oils from the leaves of SC and PG, as well as some of their terpene-enriched fractions (+V = more volatile and -V = less volatile) obtained by vacuum distillation. Both the pharmacological responses and chemical compositions were correlated.The relative contents of the oils and their fractions were evaluated by gas chromatography. Individual constituents in the oils were characterized by gas chromatography coupled to mass spectrometry. Anti-inflammatory activity was accessed in the lipopolysaccharide-induced pleurisy model, by measuring the inhibition of total leukocyte, neutrophil and eosinophil migration in the mice pleural lavage, after oil treatment with the oils at 100 mg/kg.Eosinophil migration was inhibited by SC (67%), SC (+V) (63%), PG (76%), PG (+V) (67%) and PG (-V) (74%). This efficacy was correlated with the presence of β-pinene and β-caryophyllene in the oils, a result that was reinforced by evaluating both these pure components (38 and 50% inhibition, respectively). Synergistic effects associated with the presence of α-pinene were speculated.Essential oils from SC and PG may be useful to treat inflammatory diseases by mechanisms that include the inhibition of eosinophil migration.

Galls develop in different plant organs and are induced by the activity of various organisms. Some studies have investigated the ecological interactions between species of Clusia and gall-inducing insects. The goal of our study is to characterise changes in leaf anatomy caused by the activity of gall insects in Clusia lanceolata. Additionally, we also investigated the chemical composition of volatile compounds of normal leaves and those with galls to detect possible effects on the host plants. For anatomical studies, we used botanical material fixed in FAA50. Transversal sections of the leaf blade were obtained from samples of leaves located on the third and fourth nodes from both male and female individuals. Material was studied from both sexes both with unaffected leaves and leaves containing galls. Fresh leaves of C. lanceolata were used for the extraction of volatile compounds, which were submitted to stem distillation using a modified Clevenger apparatus determining the oil yields subsequently (w/w). The unaffected leaves of female and male individuals of C. lanceolata exhibit similar anatomical structures. However, galls on leaves of both sexes show anatomical differences. The activity of the gall insect Clusiamyia nitida induces several changes in the foliar anatomy and the distribution of metabolic compounds in new tissues during gall development. However, the larvae are not able to induce significant changes in the volatile compounds of inflected leaves from male and female individuals.

Introduction Non-volatile oleoresins from neotropical Burseraceae are traditionally used for craft, technological and medicinal purposes. The crude resin is usually sold in popular markets of the forest communities. Adding value to this rainforest raw material requires establishing its composition. Objective To analyse the resin composition from different Burseraceae species and establish a minimally reproducible profile by gas chromatography, in order to parameterise its quality control. Methodology Crude oleoresin samples of 10 Protium and Trattinnickia species and a commercial blend were subjected to hydrodistillation to remove volatile compounds. The chloroform-soluble residues were methylated, analysed by GC-FID (flame ionisation detection), and individual components were identified by analysing their mass fragmentation pattern in GC-MS and comparison with data from the literature. The blend solubility was assayed in 30 non-chlorinated solvents at three different proportions. Results The resins consisted exclusively of triterpenes, showing a common predominance of four major compounds in all the samples, corresponding to α-amyrin, β-amyrin, α-amyrenone and β-amyrenone. This profile was complemented with minor amounts of the tetracyclic β-elemolic and α-elemolic acids, maniladiol, brein and other oxidised trace compounds. The better solvents for the resin were those chemically bearing more than four carbon atoms, as n-butyl acetate, 2-ethoxyethanol and isopropanol. The crude resin blend sold contained around 10% of insoluble material that was constituted by up to 70% inorganic residues mixed with humic acid derivatives, as attested by ash analysis and IR spectroscopy, respectively. Conclusion The experimental results, complemented by a general inspection of the literature, demonstrated a systematically reproducible triterpene profile in Protium and Trattinnickia species. Copyright © 2012 John Wiley & Sons, Ltd.

Two different samples of the essential oil resin harvested from two trees of Protium heptaphyllum, growing in the Brazilian southeast seashores, were obtained by hydrodistillation and analyzed by GC and GC-MS. Monoterpenes were highly predominant in both compositions of the oils. The fresher sample, harvested from the tree A, showed the predominance of myrcene (35 %), α-pinene (27 %) followed by sabinene (11 %) and β-caryophyllene (7.2 %). Terpinolene (28 %) and p-cymene (16 %) were the largest components in the trunk resin tapped by mechanical hurt-induction from the tree B. Both samples were assayed in vitro for their ability to inhibit the growth of four different of neoplasic cell. The best results were obtained from the terpinolene/p-cymene chemotype, where inhibition of 67 %, 63 % and 59 % of mouse plasmocytoma (SP2/0), neuroblastome (Neuro-2A) and monocytic cell (J774) lineages were achieved, respectively. The growth of normal cell lineage MK2 (monkey kidney epitelial cells) increased abou...

The essential oils of the leaves of Eugenia acutata , Eugenia candolleana , Eugenia copacabanensis and Myrcia splendens (Myrtaceae) from Brazil’s southeastern Atlantic Forest were obtained by hydrodistillation and analyzed by GC–MS. Oxygenated sesquiterpenes were predominant in E. copacabanensis (54.3%) and E. candolleana (50.9%) whilst hydrocarbon species predominated in E. acutata (83.4%) and M. splendens (94.5%). trans -Caryophyllene was the most abundant component in E. acutata . Isomers of guaiol and cadinol alcohols, followed by δ-elemene and viridiflorene, were the major components of the essential oil of the leaves of E. candolleana . Hydrocarbons and alcohols of the cadinane-type predominated in E. copacabanensis the most abundant being epi-cubenol (14%). M. splendens had 80% α-bisabolene in the leaf oil along with E. copacabanensis exhibited 13.7% monoterpenes. Whereas the bisabolene-rich M. splendens oil is highly similar to that of other Myrcia species reported elsewhere, the Eugenia species oils corroborated the complex array and differing abundances of terpene classes within this genus. This study generated data which may provide further comprehension of the phylogenetic relationships between Myrtaceae genera and species.

Myrtaceae is a plant family widely used in folk medicine and Syzygium and Eugenia are among the most important genera. We investigated the anti-allergic properties of an aqueous leaf extract of Syzygium cumini (L.) Skeels (SC). HPLC analysis revealed that hydrolyzable tannins and flavonoids are the major components of the extract. Oral administration of SC (25-100 mg/kg) in Swiss mice (20-25 g; N = 7/group) inhibited paw edema induced by compound 48/80 (50% inhibition, 100 mg/kg; P

Fish oil (FO), source of omega-3 fatty acids (FA), has been widely studied in the treatment of inflammatory diseases and inflammatory pain (IP). Omega-3 FA give rise to eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, metabolized to eicosanoids and converted to resolvins with important anti-inflammatory action.This study investigates the effects of oral doses of omega-3 FA from FO and concentrated fish oil (CFO) in a model of sub-chronic IP, induced by Complete Freund's Adjuvant (CFA).IP was induced by intraplantar injection of CFA into the right hind paw of Wistar rats. Three groups were pre-treated with omega-3 FA: two groups received CFO (460mg of EPA/360mg of DHA and 690mg of EPA/540mg of DHA) and one group received natural FO (460mg EPA/300mg DHA), for 7days before IP induction (pre-treatment) and 5days after induction (treatment).TNF-α levels were reduced by CFO 690 (67.9%; p0.01), CFO 460 (57.7%; p0.01), FO 460 (26.2%), compared to the augment promoted by CFA (549.7%; p0.001). Resolvin levels were increased in treated groups with respect to the CFA control group (CFO 690=3196.3%, p0.01; CFO 460=3347.1%, p0.01; FO=1653.5%).The results indicate that the tested doses reduced inflammatory pain effectively in a short pre-treatment period, through modulation of TNF-α and resolvins and that CFO presented better results than FO. Therefore, Ω-3 FA from FO can be proposed for use as complementary medicine in the treatment of painful and inflammatory diseases.

The essential oils from the oleoresin, leaf and fruit of Protium icicariba (Burseraceae), collected in the Atlantic forest were extracted by steam distillation and analysed by HRGC-MS. The resin oil was found to be entirely monoterpenoid and characterised by the predominance of p-cymene, followed by α-pinene, α-terpinolene and limonene. Monoterpenes, particularly of the terpinolene-type, predominate in the fruit oil, and germacrene-type sesquiterpenes, followed by α-copaene, γ-elemene and δ-cadinene make the greater part of the leaf oil. The leaf epicuticular wax of this species was also chemically characterised by HRGC-MS, indicating the major presence of n-nonacosane (C-29) and n-hentriacontane (C-31) together with some triterpenes. Histological sections of leaf, fruit and stem were carried out, in order to establish the microstructure present in these organs.

A new chemotype of the aromatic Verbenaceae species Lippia alba Mill. N. E. Br. from southeastern Brazil has recently been shown to have a high content of linalool in the leaf essential oil. Vegetative propagation of this chemotype was conducted at six different locations in Brazil, and the variation of the content and the optical purity of linalool in the oils were verified. Yields (0.6-0.9%, hydrodistillation), chemical composition, linalool content, and optical purity of the oils from all the plants were compared, using GC-FID, GC-MS, chiral chromatography, and retention index calculation. No plant exceeded the matrix in linalool content (46.5 to 90.7%), and the chemical profile of the oils was the same for all the samples. Purification of linalool to a content close to 100% was effected by vacuum distillation of the crude oil. Chiral analysis showed exclusively the presence of S-linalool in all the crude oils and in the distilled samples.

A mixture of the benzofuran remirol and the benzopyran isoevodionol (5.7:1) was extracted by hydrodistillation from rhizomes of Mariscus pedunculatus. They were purified by successive thin layer chromatography. Their structure elucidation were confirmed by 2D-NMR experiments. The structure of remirol was also confirmed by its hydrogenation product. Crude distillate and the pure compounds were screened for anti-inflammatory activity by the use of the mouse pleurisy model induced by lipopolysaccharide (LPS, 12.5 ng/cavity). Both the mixture and remirol showed a dose-dependent (100 and 200 mg kg-1 p. o.) capacity to inhibit neutrophil and eosinophil accumulation in the mouse pleural cavity. In contrast, no anti-inflammatory effect was observed for iso-evodionol.

The essential oils of six species of Protium (Burseraceae), collected near Manaus, Amazon, were extracted by hydrodistillation and analysed by GC – MS. Resins from four species were collected yearly, and two species were represented by more than one tree. All six oils were found to be entirely monoterpenoid in constitution and were characterized by the predominance of p-cymene and α- and β-phellandrene. The composition of fresh exudates is associated with increasing content of α-pinene and unsaturated p-menthanes. Copyright © 2000 John Wiley & Sons, Ltd.

The underivatized hexane and acetone extracts of two propolis samples (from Brazil’s southwest) were analyzed by HT-HRGC (high temperature high resolution gas chromatogrphy) and HT-HRGC coupled to mass spectrometry (HT-HRGC-MS). Several compounds, including flavonoid aglycones, phenolic acids and high molecular weight compounds could be readily characterized in the crude extracts by HTHRGC-MS. HTHRGC and HTHRGC-MS were shown to be quick and informative tools for rapid analysis of crude extracts without need for prior derivatization and purification.

The underivatized acetone and hexane fractions from propolis samples (predominant flora Citrus spp. and Vernonia polyanthes) were analyzed by HT-HRGC (high temperature high resolution gas chromatography) and HT-HRGC coupled to mass spectrometry (HT-HRGC-MS). Several compounds, including flavonoid aglycones, phenolic acids, and high molecular weight compounds were characterized in crude extracts by HT-HRGC-MS. HT-HRGC and HT-HRGC-MS were shown to be quick and informative tools for rapid analysis of crude extracts without need for prior derivatization and purification.

Pectin is a heteropolysaccharide which has been investigated for the development of colon-specific drug delivery systems. Polymers have been associated with pectin to reduce its aqueous solubility and improve the performance of drug delivery systems. Pectin–casein interaction is widely known in food research, but it has not been fully considered by pharmaceutical scientists. Thus, this study investigated the potential of casein–pectin microparticles as a drug delivery system and clarified the impact of cross-linking and drying methods on the in vitro release of indomethacin (IND) or acetaminophen (PCT) from microparticles. Microparticles were prepared by coacervation and dried by spray or spouted bed methods. Drug recovery, in vitro drug release, size, morphology, and the thermal and diffractometric properties of dried microparticles were determined. Spray-dried non-cross-linked microparticles were able to prolong IND release, and pectin was still degraded by pectinolytic enzymes. On the other hand, glutaraldehyde cross-linking prevented the enzymatic breakdown of pectin without improving IND release. Spouted bed drying reduced IND recovery from all microparticles when compared with spray drying, thus the successful spouted bed drying of microparticles depends on the chemical characteristics of both the drug and the polymer. Release data from PCT microparticles suggested that the microparticle formulation should be improved to bring about a more efficient delivery of water-soluble drugs. In conclusion, casein–pectin microparticles show great potential as a drug delivery system because casein reduces the water solubility of pectin. The drying method and cross-linking process had significant effects on the in vitro performance of these microparticles.

This study investigated the leishmanicidal activity againstLeishmania amazonensisof four commercial oils fromCopaiferaspp. named as C1, C2, C3, and C4, the sesquiterpene and diterpene pools obtained from distilling C4, and isolatedβ-caryophyllene (CAR). Copaiba oils chemical compositions were analyzed by gas chromatography and correlated with biological activities. Diterpenes-rich oils C2 and C3 showed antipromastigote activity. Sesquiterpenes-rich C1 and C4, and isolated CAR presented a dose-dependent activity against intracellular amastigotes, with IC50sof 2.9 µg/mL, 2.3 µg/mL, and 1.3 µg/mL (6.4 µM), respectively. Based on the highest antiamastigote activity and the low toxicity to the host cells, C4 was steamdistillated to separate pools of sesquiterpenes and diterpenes. Both pools were less active againstL. amazonensisand more toxic for the macrophages than the whole C4 oil. The leishmanicidal activity of C3 and C4 oils, as well as C4 fractions and CAR, appears to be independent of nitric oxide production by macrophages. This study pointed outβ-caryophyllene as an effective antileishmanial compound and also to its role as potential chemical marker in copaiba oils or fractions derived thereof, aiming further development of this rainforest raw material for leishmaniasis therapy.

In continuation of a search for new sources of aromatic material from the Burseraceae family (Siani et al., 1999a; Siani et al., 1999b; Ramos et al., 2000) we investigated the essential oil of the trunk exudate of Trattinickia rhoifolia Willd (Burseraceae) (Daly, 1999). Two different samples of the resin were collected (April 98 and January 99) from the same tree, in the Ducke Biological Reserve, 21 km from Manaus. The tree was catalogued by the Projeto Flora in the National Institute of Research of Amazonia—INPA, and identified by J.E.L.S. Ribeiro. A voucher specimen has been deposited at the INPA Herbarium under no. 2848.

The composition of essential oils from two sources of oleoresins of Protium heptaphyllum were analyzed by GC/MS and Retention Index. The composition of the oils varied according to fresh or aged resin. The oil of fresh resin was rich in α-terpinene (18.0%), p-cymene (36.0%) and γ-terpinene (12.0%), whereas the aged resin oil contained p-cymene (11.0%), terpinolene p-cymen-8-ol (11.0%) and dillapiole (16.0%).

Myrtaceae is a plant family widely used in folk medicine and Syzygium and Eugenia are among the most important genera. We investigated the anti-allergic properties of an aqueous leaf extract of Syzygium cumini (L.) Skeels (SC). HPLC analysis revealed that hydrolyzable tannins and flavonoids are the major components of the extract. Oral administration of SC (25-100 mg/kg) in Swiss mice (20-25 g; N = 7/group) inhibited paw edema induced by compound 48/80 (50% inhibition, 100 mg/kg; P

Methanol extracts of propolis from six different places, five in Rio de Janeiro state and one in Sao Paulo state, both in the Southeast of Brazil, were investigated using high-temperature high-resolution gas chromatography (HT-HRGC) and HT-HRGC-mass spectometry. The main purpose of the study was to establish the applicability of HT-HRGC as an analytical method for systematic studies of polar propolis fractions. Several compounds, including carbohydrates, phenolic acid derivatives, and high molecular weight compounds (e.g., wax esters of long chain fatty alcohols) could be readily characterized in the crude extracts by HT-HRGC-MS. HT-HRGC and HT-HRGC-MS were shown to be quick and informative tools for rapid analysis of crude polar extracts without cleanup.

Synopsis The aim of this work was to evaluate several plant extracts with regard to ultraviolet absorption spectra in view of a possible application as antisolar agents. Liquid and dry extracts of Hamamelis virginiana, Matricaria recutita, Aesculus hippocastanum, Rhamnus purshiana and Cinnamomum zeylanicum were prepared by repercolation, maceration and microwave oven extraction. UVB absorption spectra (290-320nm) were obtained and the solar protection factors (SPF) of these preparations were determined by a spectrophotometric method. The results showed that after incorporation to a 2% solution of the synthetic sunscreen octylmethoxycinnamate, the extracts showed an intensification in SPF values, suggesting that this can be an interesting method to intensify SPF. In addition, these extracts can contribute their emollient and moistening properties to the product. These are important characteristics for protecting skin against exposure to the sun.

Considerando que a obtenção de uma formulação farmacêutica pode ser melhor planejada se algumas condições biológicas inerentes à cavidade oral forem contempladas e que os sistemas de liberação modificada podem ser ferramentas para o controle de doenças orais, o objetivo do trabalho é obter uma formulação contendo clorexidina, avaliando seu efeito sobre os mesmos. Este trabalho foi dividido em duas etapas, sendo que a primeira envolveu a obtenção da formulação mais adequada aos experimentos biológicos e a segunda avaliou seus efeitos sobre biofilmes patogênicos orais. Assim, formulações com diferentes concentrações de clorexidina foram preparadas e avaliadas visualmente em relação à sua integridade física nas condições de crescimento dos biofilmes, seguido de ensaio de liberação em meio estático utilizando tampão como meio de dissolução (meio de dissolução convencional). A formulação selecionada foi submetida a ensaios de liberação em meio estático contendo os diferentes caldos de cultura bacterianos como meio de dissolução. Na segunda etapa do trabalho, o efeito da formulação selecionada foi avaliado em biofilmes cariogênicos de Streptococcus mutans ou biofilmes periodontopatogênicos de Porphyromonas gingivalis. Biofilmes de S. mutans (n=6) ou P. gingivalis (n=3) foram formados em caldos de cultura sob lâminas de vidro por 6 e 3 dias, respectivamente, sendo expostos a um dos seguintes tratamentos: 1) Formulação contendo 92% de quitosana e 8% de hidroxipropil metilcelulose (CV, como controle do veículo) ou 2) Formulação contendo 82% de quitosana, 8% de hidroxipropil metilcelulose e 10% de clorexidina (CHX, grupo experimental). Um grupo sem exposição à formulação foi incluído como controle negativo (CN). Após o período experimental, a viabilidade bacteriana, o pH dos biofilmes e a quantificação de clorexidina liberada para os caldos de cultura foram determinados. Os dados foram analisados estatisticamente por teste de Tukey-Kramer e Tukey, sendo o nível de significância estabelecido em 5%. Os resultados sugerem que ainda que a liberação de clorexidina da formulação nos caldos de cultura de S. mutans e P. gingivalis tenha sido menor se comparada ao meio de dissolução convencional (p0,05), mas apresentaram quedas de pH maiores se comparados à CHX (p0,05). Para P. gingivalis também houve mais morte celular nos biofilmes expostos à CHX (p 0.05), but showed higher pH drops when compared to CHX (p 0.05). For P. gingivalis, there was higher cell death in the biofilms exposed to CHX (p

O câncer de bexiga (CB) destaca-se como o segundo tipo de doença mais comum do trato geniturinário e o décimo com maior incidência no Brasil, sendo responsável por cerca de 4020 óbitos por ano. Os dados refletem a baixa eficácia dos tratamentos disponíveis. Assim, novas moléculas e sistemas de liberação estão sendo investigados para terapia intravesical do CB. Nesta linha, as oleorresinas de Copaifera e compostos fenólicos como a curcumina (CM) e demetoxicurcumina (DCM) presentes no extrato de Curcuma longa, têm sido alvo de pesquisas por apresentarem propriedades antitumoral, antiproliferativa e proapoptótica. Contudo, a administração desses compostos em sua forma livre torna-se inviável devido à sua alta lipossolubilidade e instabilidade. Dessa forma, o uso de oleorresina de Copaifera duckei (O-Cd) no preparo de carreadores lipídicos nanoestruturados (CLN) para a encapsulação do extrato de C. longa (ECL) poderia ser uma estratégia terapêutica eficaz, além de transpor algumas barreiras da terapia intravesical como a baixa permeação de moléculas no urotélio e seu baixo tempo de residência na bexiga. Assim, os objetivos do presente estudo foram preparar e caracterizar CLN (com O-Cd) e CLN imperfeito (sem O-Cd, CLNimp) para a encapsulação de ECL, e avaliar a citotoxicidade desses sistemas carreadores em células de CB. Os CLN e CLNimp, preparados pelo método de emulsão à quente e sonicação, foram caracterizados quanto ao seu diâmetro, índice de polidispersão (PdI), potencial zeta (PZ), cristalinidade, eficiência de encapsulação (EE) por CLAE, capacidade de encapsulação (CE), e morfologia. Além disso, avaliou-se a citotoxicidade dos CLNs com e sem ECL em células RT4 e foram conduzidos estudos de uptake celular. Obteve-se carreadores com diâmetro ao redor de 200 nm, baixo PdI (?0,3) e PZ negativo (~-20 mV). O ECL foi caracterizado e quantificado por CLAE, sendo verificado uma pureza de 97,90% para CM e 99,29% para DCM. Ambas moléculas foram encapsuladas nos dois CLNs com alta EE (~70%). A encapsulação do ECL não alterou de forma significativa as propriedades físicas dos CLNs. Os dois tipos de CLNs apresentaram baixo índice de recristalização (~60%), forma esférica e superfície lisa. Os CLN e CLNimp foram estáveis no período de 420 e 270 dias, respectivamente. Já os CLN-ECL e CLNimp-ECL foram estáveis por 90 dias. O IC50 dos CLN sem ECL (0,019x1012 partículas/mL) foi menor que o IC50 dos CLNimp sem ECL (>0,061x1012 partículas/mL). O encapsulamento do ECL pelos CLNs não alterou significativamente seu IC50 nos períodos de incubação avaliados. Já o encapsulamento do ECL pelos CLNimp diminuiu o IC50 (19,15 ?g/mL) (0,047x1012 partículas/mL) em relação ao CLNimp vazio. Observou-se maior uptake celular dos carreadores com ECL em relação ao ECL livre. Além disso, foi observada fragmentação do núcleo das células que internalizaram CLN-ECL. As interessantes propriedades físicas e biológicas apresentadas pelos sistemas CLN vazio e CLNimp-ECL indicam seu potencial terapêutico e os tornam promissores para futura terapia do câncer de bexiga. Bladder cancer (BC) stands out as the second most common genitourinary disease type and the tenth one with the highest incidence in Brazil, being responsible for about 4020 deaths per year. The data reflect the low efficacy of the available treatments. Thus, new molecules and release systems are being investigated for intravesical BC therapy. In this line, Copaifera oleoresins and phenolic compounds such as curcumin (CM) and demethoxycurcumin (DCM) present in Curcuma longa extract, have been the subject of research because they have antitumor, antiproliferative and proapoptotic properties. However, administration of these compounds in their free form becomes infeasible due to their high liposolubility and instability. Therefore, the use of Copaifera duckei oleoresin (Cd-O) in the preparation of nanostructured lipid carriers (NLC) for the encapsulation of C. longa extract (CLE) could be an effective therapeutic strategy, in addition to transposing some barriers of intravesical therapy such as the low permeation of molecules in the urothelium and its low residence time in the bladder. Thus, the aims of the present study were to prepare and characterize NLC (with Cd-O) and imperfect NLC (without Cd-O, NLCimp) for the encapsulation of CLE, and to evaluate the cytotoxicity of these carrier systems in BC cells. The NLC and NLCimp, prepared by the hot emulsion and sonication method, were characterized by their diameter, polydispersity index (PdI), zeta potential (ZP), crystallinity, encapsulation efficiency (EE) by HPLC, encapsulation capacity (EC) and morphology. In addition, the cytotoxicity of NLCs with and without CLE was evaluated in RT4 cells and cell uptake studies were conducted. Carriers with diameter about 200 nm, low PdI ( 0.061x1012 particles/mL). The encapsulation of CLE by NLCs did not significantly alter its IC50 in the incubation periods evaluated. The encapsulation of CLE by the NLCimp decreased its IC50 (19.15 ?g/mL) (0.047x1012 particles/mL) in relation to the empty CLNimp. It was observed higher cellular uptake of the carriers with CLE in relation to free CLE. In addition, fragmentation of the nucleus of the cells that internalized NLC-CLE was observed. The interesting physical and biological properties presented by empty NLC and NLCimp-CLE systems indicate their therapeutic potential and make them promising for future bladder cancer therapy

O perfil de liberação e o efeito de um sistema de liberação prolongada contendo metronidazol, antimicrobiano prescrevido para o tratamento da periodontite, foram avaliados na presença de biofilmes supra e subgengivais, representados respectivamente pelas bactérias Streptococcus mutans e Porphyromonas gingivalis. Os biofilmes foram crescidos e expostos ao sistema de liberação prolongada contendo metronidazol (MDZ) ou ao controle de veículo da formulação (CV), composto de monoglicerídeos e monoesterato de sorbitano. Biofilmes não tratados foram utilizados como controle negativo (CN). Os biofilmes e os meios de cultura de S. mutans foram coletados após a primeira exposição aos tratamentos nos tempos 24, 48, 72 e 96 horas enquanto para biofilmes de P. gingivalis os tempos foram 24, 48 e 72 horas. Após coleta, os biofilmes foram analisados em relação à quantificação de fármaco e viabilidade bacteriana (biofilmes de S. mutans: n=3; biofilmes de P. gingivalis: n=6). Biofilmes de S. mutans também foram avaliados em relação à acidogenicidade. Nos biofilmes supragengivais, a quantificação de MDZ nas primeiras 24 horas foi de 7% em relação à concentração inicial de fármaco na formulação, permanecendo em torno de 1% para os demais tempos. O teor de MDZ liberado da formulação reduziu a viabilidade bacteriana no tempo 24 horas e diminuiu a acidogenicidade dos biofilmes por 48 horas em relação aos grupos CV e NC (p0,05). O grupo CV apresentou menor viabilidade bacteriana se comparado ao grupo CN (p 0.05). The VC group presented lower number of viable bacteria than NC (p

As infecções causadas pelo Herpes simplex virus, tipo 1 e 2 (HSV-1 e HSV-2) são consideradas problema de saúde pública no mundo, com prevalência em dois terços da população mundial. A doença é caracterizada pelo aparecimento de vesículas que ocasionam dor e constrangimento ao portador, devido à aparência desagradável que apresentam. As infecções que levam a lesões orofaciais são geralmente ocasionadas pelo HSV-1, enquanto as genitais, pelo HSV-2. O tratamento de escolha baseia-se no uso de antivirais em géis ou pomadas, entretanto, recidivas são comuns e dependentes do estado imunológico do indivíduo, além de exposição a fatores hormonais ou ambientais. Os anestésicos locais diminuem a dor ocasionada pela lesão além de mostrarem ter propriedades antivirais. A forma semissólida facilita a aplicação e a sua transformação em uma película fina, após curto período de tempo, favorece a manutenção da formulação no local, com aspecto final mais discreto quando comparada às opções disponíveis. O objetivo desse trabalho foi desenvolver e avaliar in vitro e in vivo formulações contendo anestésicos para o tratamento do herpes labial. Para tanto, duas formulações semissólidas formadoras de filme foram desenvolvidas e avaliadas (F1 e F1T), contendo HPMC K100, lidocaína (LIDO) e prilocaína (PRILO) combinados a adjuvantes, na presença (F1T) ou não (F1) do promotor de absorção Transcutol®. A mistura de PRILO e LIDO acarretou na formação de uma mistura eutética (ME). Para quantificação dos fármacos a partir da formulação, um método analítico por CLAE (Cromatografia Líquida de Alta Eficiência) foi revalidado e atendeu aos parâmetros preconizados na literatura. A recuperação dos fármacos a partir da pele suína foi satisfatória. As formulações apresentaram homogeneidade na distribuição do conteúdo. A formação do filme in situ foi avaliada e ocorreu em aproximadamente 20 minutos. A adição do promotor de absorção aumentou a concentração dos fármacos no líquido receptor no experimento de permeação passiva in vitro, utilizando pele de orelha suína. E a quantidade de PRILO e LIDO quantificada no estrato córneo foi maior para a formulação sem o Transcutol®. As análises reológicas de cisalhamento contínuo e oscilatório classificaram as formulações como fluídos newtonianos e a F1T apresentou maior estruturação em relação a F1. A avaliação in vitro da multiplicação viral sugere atividade virucida para ambos os fármacos, com proteção a infecção superior a 80% para as concentrações de fármaco avaliadas. As formulações não apresentaram irritação dérmica ao exame macroscópico. O teste in vivo de eficácia comprovou a capacidade dos anestésicos em regredir as lesões causadas por HSV-1. Sendo assim, as formulações propostas mostraram-se boas alternativas ao tratamento de lesões labiais causadas pelo HSV-1. Análises estatísticas adequadas foram realizadas. Infections by Herpes Simplex virus, type 1 and 2 (HSV 1 e HSV-2) are a public health problem in the world with prevalence of two thirds of the mundial population. This disease is characterized by painful vesicles and embarrassment because of wound appearance. Orofacial wounds are mostly caused by HSV-1 and genital wounds are by HSV-2. Nowadays, current treatment is based on the use of antiviral in cream and ointments but, recurrences are very common and depend of individual state of immunological system besides environmental factors. Topical anesthetics minimize the pain and have shown antiviral properties. The semisolid composition improves application and its transformation into a thin film, after a short period of time, favors the maintenance at the site with good final aspect compared to the currently available treatment options. In this research, we developed and evaluated in vitro and in vivo compositions containing topical anesthetics for treatment of labials herpes. The compositions were prepared using HPMC K100, lidocaine (LIDO) and prilocaine (PRILO) combined with adjuvants, with permeation promoter Transcutol® (F1T) or not (F1). The mix of PRILO and LIDO results in eutectic mixture. For quantified drugs in compositions an analytical method by HPLC (High Performance Liquid Chromatography) was revalidated with satisfactory parameters, as well the recovery of drug on porcine skin. The formulations were with homogeneous content and film formation occurs in approximately 20 minutes. The addition of Transcutol® results in more PRILO and LIDO in receptor medium for F1T at in vitro passive permeation study. The quantify of drugs in corneum layer is mayor in F1 than F1T. Rheological analyses shown the compositions are newtonians fluids and F1T has more cohesion than F1. Evaluation of in vitro viral multiplication suggests activity before virus infects host cells, with 80% of protection of infection for evaluated concentrations. None composition shown dermal irritation in macroscopical examination. The in vivo efficacy study proves the anesthetics capacity of regress the HSV-1 wounds. Therefore, the developed compositions are good alternatives for labialis herpes caused by HSV-1.

A pele desempenha um importante papel na homeostasia e é uma via muito utilizada na administração de fármacos. O envelhecimento compromete sua função protetora e contribui para o aparecimento de sintomas como a rugosidade, flacidez, secura, prurido, hiperpigmentação dificuldade de cicatrização e a incidência de melanomas. Os raios UV são a principal causa do envelhecimento devido à geração de espécies reativas de oxigênio (ROS) e peroxidação lipídica. Os danos causados pelos ROS podem ser mitigados pelo uso de antioxidantes tópicos como a curcumina e timol (TML). No entanto, a baixa permeabilidade e estabilidade são fatores limitantes para sua aplicação. Para superar esses desafios, ECUR e TML podem ser encapsulados em nanopartículas, como os carreadores lipídicos nanoestruturado (CLN). Assim, os objetivos desse projeto foi desenvolver um CLN estável com manteiga Illipê, óleo de calêndula e TPGS para co-encapsular ECUR e TML e incorporação em uma formulação tópica. O padrão analítico dos curcuminóides foi isolado de amostra comercial, identificado por LC-MS e RMN e a metodologia analítica para quantificação CUR e TML foi validada. Os CLNs foram produzidos por emulsão quente e sonicação e otimizados utilizando a ferramenta de DoE Box-Behnken. Foram caracterizados em relação às propriedades físicas, citotoxicidade em cálulas HaCaT e atividade antioxidante. O CLN otimizado apresentou tamanho em torno de 130 nm, índice de polidispersão abaixo de 0,200 e potencial zeta negativo. A eficiência de encapsulação foi acima de 97% tanto para TML quanto CUR. CLN com e sem ativos demonstrou baixo índice de recristalização e forma esférica caracterização por DSC e AFM e não foram citotóxicos para células HaCaT, mostrando viabilidade celular superior a 80% na concentração 1,48x1012 partículas/ml, correspondente a 0,8 ?g/ml de TML/CUR co-encapsulados. Além disso, TML/CUR livre e co-encapsulado mostrou alta atividade antioxidante em ensaios in vitro para inibição de xantina oxidade e auto-oxidação do pirrogalol, indicando sua potente capacidade de inibição da enzima superóxido dismutase mesmo quando encapsulados. O emulgel tópico foi desenvolvido e apresentou estabilidade física após a incorporação do CLN no teste de estabilidade acelerada. O perfil reológico do emulgel demonstrou característica plástica e reoplexa e se manteve estável com o tempo e sob as variações de temperatura de analise, indicando alta estabilidade física da formulação. O perfil de liberação de ECUR e TML indicou cinética de liberação correspondente ao modelo de Higuchi sem influência da co-encapsulação na liberação da CUR e com liberação mais lenta do TML encapsulado. Os dados de permeação sugerem que os ativos possam permear na pele. A extensão e a intensidade da permeabilidade dos ativos promovida pelo CLN determinarão a aplicabilidade da formulação que poderá ser interessante para o desenvolvimento de um produto cosmético, dermocosmético, ou mesmo para um medicamento transdérmico. Em conclusão, o CLN desenvolvido apresentou características físicas e biológicas interessantes para a aplicação tópica. Skin plays an important role in homeostasis and it is an important route for drug delivery. Aging compromises its protective function and contributes to the appearance of symptoms such as roughness, sagging, dryness, pruritus, hyperpigmentation, scarring deficient and the incidence of melanomas. UV rays are the main cause of photo-aging due of ROS generation and lipid peroxidation. ROS damage can be mitigated by using topical antioxidants as curcumin and thymol. However, low permeability and physico-chemical stability are disadvantages for its clinical application. To overcome these challenges, ECUR and TML can be encapsulated in nanoparticles such as nanostructured lipid carriers (NLC). This project aimed to develop a stable NLC composed by Illipê butter, calendula oil and TPGS to co-encapsulate ECUR and TML, followed by the NLC incorporation into a cosmetic formulation. Curcuminoid\'s analytical standard was isolated from commercial sample, identified by LC-MS and NMR, and the analytical methodology for quantification CUR and TML was validated. NLCs were produced by sonication emulsification method and their development was designed by Box-Behnken DoE and characterized in relation to physical properties, HaCaT cell cytotoxicity and antioxidant activity. Optimized NLC with size around 130 nm, polydispersity index (PdI) below 0.200 and negative zeta potential was obtained. The encapsulation efficiency (EE%) was above 97% for both TML and CUR. Empty and co-encapsulated NLC showed in DSC and AFM characterization low recrystallization index and spherical shapes and were not cytotoxic for HaCaT cells, showing cell viability over 80% at the maximum concentration evaluated (1.48x1012 particles / ml, wich corresponds to 0.8 ?g / ml co-encapsulated TML / CUR). Furthermore, free and co-encapsulated TML / CUR showed high antioxidant activity in in vitro assays due inhibition of xanthine oxidation and pyrogallol autoxidation, indicating their potent superoxide dismutase inhibition even when co-encapsulated. The cosmetic emulgel was developed and physically stable after NLC incorporation in accelerated stability. Emulgel rheological profile showed a plastic and rheopexy characteristic, which remained storage stability. The rheological profile did not change due variations in the temperature analysis, indicating good formulation physical stability and cosmetic acceptability. The release profile of ECUR/TML release kinetics corresponded to the Higuchi model without influence of co-encapsulation with slower release of the encapsulated TML. Permeation data suggest that the active substances may permeate deep into skin. The actives permeability extent and intensity promoted by CLN will determine the applicability of the formulation that can be interest for the development of a cosmetic, dermocosmetic product or even for a transdermal drug. In conclusion, the developed CLN presented interesting physical and biological characteristics for topical application.