Copaifera langsdorffii Oleoresin-Loaded Nanostructured Lipid Carrier Emulgel Improves Cutaneous Healing by Anti-Inflammatory and Re-Epithelialization Mechanisms.

The skin is essential to the integrity of the organism. The disruption of this organ promotes a wound, and the organism starts the healing to reconstruct the skin. Copaifera langsdorffii is a tree used in folk medicine to treat skin affections, with antioxidant and anti-inflammatory properties. In our study, the oleoresin of the plant was associated with nanostructured lipid carriers, aiming to evaluate the healing potential of this formulation and compare the treatment with reference drugs used in wound healing. Male Wistar rats were used to perform the excision wound model, with the macroscopic analysis of wound retraction. Skin samples were used in histological, immunohistochemical, and biochemical analyses. The results showed the wound retraction in the oleoresin-treated group, mediated by α-smooth muscle actin (α-SMA). Biochemical assays revealed the anti-inflammatory mechanism of the oleoresin-treated group, increasing interleukin-10 (IL-10) concentration and decreasing pro-inflammatory cytokines. Histopathological and immunohistochemical results showed the improvement of re-epithelialization and tissue remodeling in the Copaifera langsdorffii group, with an increase in laminin-γ2, a decrease in desmoglein-3 and an increase in collagen remodeling. These findings indicate the wound healing potential of nanostructured lipid carriers associated with Copaifera langsdorffii oleoresin in skin wounds, which can be helpful as a future alternative treatment for skin wounds.

Evaluation of anti-inflammatory activity of kaurenol: Experimental evaluation and mechanistic insights.

BACKGROUND: Kaurenol, a diterpene alcohol found in Copaifera langsdorffii Desf. (known as "copaiba"), is historically used in traditional medicine for inflammatory conditions. OBJECTIVES: This study aims to comprehensively assess the potential anti-inflammatory and antinociceptive properties of kaurenol. METHODS: To this end, the following experiments were conducted to evaluated toxicity: locomotor performance and acute toxicity; nociception: acetic acid-induced writhing and formalin-induced antinociception; and anti-inflammatory activity: carrageenan and dextran-induced paw edema at 10, 20, and 40 mg/kg, and measurement of nitric oxide (NO), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10) in macrophages at 1, 3, and 9 µg/ml. RESULTS: Kaurenol did not show significant locomotor changes, acute toxicity, and central analgesic activity in the first phase of formalin test at dosages tested. Kaurenol showed 53%, 64%, 64%, and 58% of inhibition in the acetic acid-induced writhing, second phase of formalin test, carrageenan and dextran-induced paw edema, respectively. CONCLUSION: The anti-inflammatory activity was associated with the regulation of NO release and probably with the regulation of mediators, such as serotonin and prostaglandin in vascular permeability, as well as by being associated with the regulation of IL-6 and IL-10. Kaurenol display anti-inflammatory activity but has no analgesic activity.

Mathematical and numerical analyses of cellular, molecular and angiogenic parameters of a rat skin wound healing model.

The inflammatory phase is an important event in the skin wound healing process. The deposition of granulation tissue in the wound bed and the rebuilding of the vascular network occur as inflammation diminishes. An angiogenic component in the formation of granulation tissue is the secretion of vascular endothelial growth factor, which assists in the chemotaxis, proliferation, and replication of fibroblasts. In this paper, we develop a mathematical model of skin wound healing angiogenic factors based on inflammatory cells (macrophages and neutrophils) and mediators (interleukin 6 and interleukin 10). We highlight the importance of this process in vascular endothelial growth factor release and in the formation of new capillary tips. We used a mathematical model of partial differential equations based on the reaction-diffusion-advection equations. In order to calibrate the parameters, we considered an in vivo model composed by four treatments: hydroalcoholic extract and oil-resin of Copaifera langsdorffii at 10% concentration, collagenase, and Lanette cream. Using the laboratory data for the wound edge, our mathematical model estimated the values of vascular endothelial growth factor concentration, and tips density in the center of the wound with a maximum error of 2.9%, and predicted healing time required for each treatment. The region of viability for the parameters, in the proposed model, was found through numerical simulations from the Interleukin 6 and 10 dysregulation and we obtained that, among the parameters analyzed, the greatest influencer in the dynamics of the system is the one, which represents the production of Interleukin 10 during phagocytosis.

Chemical composition variation of essential oils of Copaifera langsdorffii Desf. from different vegetational formations.

This study aims to provide information about the chemical profile of the essential oil from C. langsdorffii resin in areas of Cerradão, Carrasco, and Humid Forest. In order to obtain the essential oils, oil-resin was submitted to hydrodistillation process, and the chemical components were analyzed by gas chromatography coupled to mass spectrometry (GC/MS). Twenty-six constituents were identified in the essential oil of C. langsdorffii resin, of which 12 were present in Cerradão, 15 in Carrasco and 18 in Humid Forest, with ß-bisabolene, caryophyllene oxide, γ-muurolene, α-caryophyllene and ß-caryophyllene common to the three phytophysiognomies. Among the studied areas, the resin collection varied from 2.652 g to 20.263 g, while the essential oil yield varied from 2.216 % to 11.764 %. A concentration range of 0.60 % to 84.57 % was recorded among the compounds in the three study areas. There was variation in chemical composition both among phytophysiognomies and among individuals from the same location, where ß-caryophyllene showed to be the majority for all areas studied, being present in all individuals from Cerradão, Carrasco and Humid Forest.

Preventive activity of Copaifera langsdorffii Desf. leaves extract and its major compounds, afzelin and quercitrin, on DNA damage in in vitro and in vivo models.

Copaifera langsdorffii Desf. is a plant found in South America, especially in Brazil. Oleoresin and the leaves of this plant is used as a popular medicinal agent. However, few studies on the chemical composition of aerial parts and related biological activities are known. This study aimed to examine the cytotoxic, genotoxic, and antigenotoxic potential of C. langsdorffii aerial parts hydroalcoholic extract (CLE) and two of its major compounds afzelin and quercitrin. The cytotoxic and antigenotoxic potential of CLE was determined as follows: 1) against genotoxicity induced by doxorubicin (DXR) or methyl methanesulfonate (MMS) in V79 cells; 2) by direct and indirect-acting mutagens in Salmonella typhimurium strains; and 3) by MMS in male Swiss mice. The protective effects of afzelin and quercitrin against DXR or MMS were also evaluated in V79 and HepG2 cells. CLE was cytotoxic as evidenced by clonogenic efficiency assay. Further, CLE did not induce a significant change in frequencies of chromosomal aberrations and micronuclei; as well as number of revertants in the Ames test demonstrating absence of genotoxicity. In contrast, CLE was found to be antigenotoxic in mammalian cells. The results also showed that CLE exerted inhibitory effect against indirect-acting mutagens in the Ames test. Afzelin and quercitrin did not reduce genotoxicity induced by DXR or MMS in V79 cells. However, treatments using afzelin and quercitrin decreased MMS-induced genotoxicity in HepG2 cells. The antigenotoxic effect of CLE observed in this study may be partially attributed to the antioxidant activity of the combination of major components afzelin and quercitrin.

Products of Dental Use Containing Copaiba Oil-resin: Technological Prospecting Based on Patents.

BACKGROUND: Copaiba oil-resin has been widely used and is especially found in neotropical regions, for which several pharmacological activities have been documented over the years. Prospective studies in intellectual property banks are important to increase competitiveness and thus generate new products in various research areas. OBJECTIVE: A prospective study was carried out on patents of products containing copaiba oil-resin for dental use in intellectual property banks. METHODS: The research was conducted with patent searches in six intellectual property banks of the world. Relevant information about the invention in the patent document was collected, processed and described. RESULTS: The search found 9 patents using copaiba resin oil-resin in dental products. The National Institute of Industrial Property (INPI-Brazil) had the highest number of deposits (5), followed by Espacenet (2) and Free Patents (2). C. Langsdorffii was highlighted as the most widely used species in the products and deposits of vehicles in formulations (3). All the patents in the search are A61K code for medical, dental or hygienic purposes. CONCLUSION: Most of the found patents are related to the area of Microbiology, specifically with application in Cariology. Brazil is represented by the INPI and presented the highest number of patent applications when compared to other intellectual property banks.

Evaluation of cytoprotective effects of compounds isolated from Copaifera langsdorffii Desf. against induced cytotoxicity by exposure to methylmercury and lead.

Copaifera langsdorffii L. is one of the most known medicinal species in Brazil. Its leaves are rich in phenolic compounds with potential biological activities as an antioxidant and chelating agent. This paper reports the isolation of four compounds from the hydroalcoholic extract of the leaves of C. langsdorffii and the investigation of their possible cytoprotective effects against heavy metal poisoning. Quercitrin (1), afzelin (2), 3,5-di-O-(3-O-methyl galloyl) quinic acid (3) and 4,5-di-O-(3-O-methyl galloyl) quinic acid (4), were associated with toxic doses of methylmercury and lead and evaluated by Alamar blue cell viability assays in HepG2 and PC12. The compounds displayed significant cytoprotective effect for the HepG2 cell line against both metals. Compounds 1-4 did not protect PC12 cells against methylmercury induced-cytotoxicity, but at lower concentrations, they protected against lead induced-cytotoxicity. The evaluated compounds showed a promising cytoprotection effect against exposure to heavy metals and should be further investigated as protective agents.

Antifungal activity of Copaíba resin oil in solution and nanoemulsion against Paracoccidioides spp.

Paracoccidioidomycosis (PCM) is a disease caused by fungi of the genus Paracoccidioides. The disease is responsible for high rates of premature deaths and socioeconomic repercussions. The limitations of antifungal agents against PCM have motivated the search for new compounds. In our ongoing exploration of Cerrado plants as potential sources of new antifungal agents, we selected Copaifera langsdorffii oil (Copaíba resin oil) in order to explore its bioactive potential and test a formulation to increase oil stability and solubilization employing Pluronic F-127 to obtain the nanoemulsion of the oil. We aim at testing both Copaíba resin oil and its nanoemulsion against four species of the Paracoccidioides genus. We performed cytotoxicity test in Balb/C3T3 cells, hemolytic activity and interaction of Copaíba resin oil and Copaíba resin oil nanoemulsion (CopaPlu) with the antifungal agents such as amphotericin B, co-trimoxazole, and itraconazole. Moreover, the Copaíba resin oil was analyzed by mass spectrometry to identify its chemical profile. Eventually, a new methodology to prepare the nanoemulsion is presented. The Copaíba resin oil and CopaPlu nanoemulsion inhibited Paracoccidioides sp. growth efficiently, and no cytotoxicity or hemolytic effect was observed at minimum inhibitory concentration (MIC). When combined with amphotericin B, Copaíba resin oil and its nanoemulsion showed an additive effect with reduction of MIC values. The Copaíba resin oil and CopaPlu nanoemulsion is a promising antifungal agent against Paracoccidioides.

Differentiation of Essential Oils Using Nanofluidic Protein Post-Translational Modification Profiling.

Current methods for the authentication of essential oils focus on analyzing their chemical composition. This study describes the use of nanofluidic protein post-translational modification (PTM) profiling to differentiate essential oils by analyzing their biochemical effects. Protein PTM profiling was used to measure the effects of four essential oils, copaiba, mandarin, Melissa, and turmeric, on the phosphorylation of MEK1, MEK2, and ERK1/2 in the MAPK signaling pathway; Akt and 4EBP1 in the pI3K/Akt/mTOR signaling pathway; and STAT3 in the JAK/STAT signaling pathway in cultured HepG2 cells. The gain or loss of the phosphorylation of these proteins served as direct read-outs for the positive or negative regulatory effects of essential oils on their respective signaling pathways. Furthermore, protein PTM profiling and GC-MS were employed side-by-side to assess the quality of the essential oils. In general, protein PTM profiling data concurred with GC-MS data on the identification of adulterated mandarin, Melissa, and turmeric essential oils. Most interestingly, protein PTM profiling data identified the differences in biochemical effects between copaiba essential oils, which were indistinguishable with GC-MS data on their chemical composition. Taken together, nanofluidic protein PTM profiling represents a robust method for the assessment of the quality and therapeutic potential of essential oils.

Quantitative analysis of phenolic metabolites in Copaifera langsdorffii leaves from plants of different geographic origins cultivated under the same environmental conditions.

INTRODUCTION: Galloylquinic acid derivatives and flavonoids are the main phenolic metabolites found in Copaifera langsdorffii leaves (Leguminosae, Detarioideae), a medicinal plant with potential therapeutic application in the treatment of kidney stones. The factors that affect metabolite production in this plant species are not well understood but may include environmental and genetic factors. OBJECTIVE: To quantify the variation in metabolite production over a 12-month period for 10 groups of C. langsdorffii cultivated under the same environmental conditions. METHODS: Copaifera langsdorffii seeds were collected from 10 different regions in southeast, Brazil and grown in the same field. HPLC-UV was used to quantify nine galloylquinic acid derivatives and two flavonoids in leaf samples from mature trees. Climate data for humidity, radiation, precipitation and temperature were provided by the National Institute of Meteorology, Brazil. Multivariate analyses were performed to correlate chemical and environmental variables. RESULTS: The overall effect of environmental factors on the production of phenolic metabolites was uniform among C. langsdorffii groups. Chemical variation between groups was present, but small, and probably due to differences in their genetics and physiology. Seasonal changes influenced the production of the major phenolic metabolites, with increases in temperature and radiation levels favouring metabolite production. CONCLUSION: When C. langsdorffii trees are cultivated in the same environment, the production of the major secondary metabolites found in their leaves is very similar quantitatively, varying based on geographic location of original population and seasonal changes. This favours the standardisation of plant raw material for the production of a phytomedicine.

Eficiência de óleos essenciais na qualidade sanitária e fisiológica em sementes de feijão-fava (Phaseolus lunatus L. ) / Efficiency of essential oils in the sanitary and physiological quality of lima bean seeds (Phaseolus lunatus L. )

RESUMO A espécie Phaseolus lunatus L., conhecida popularmente como feijão-fava constitui uma das alternativas de renda e alimentação para a população da região Nordeste do Brasil. Um dos problemas enfrentados são as doenças que provocam grandes perdas na produção de sementes. O presente trabalho objetivou avaliar a eficiência dos óleos essenciais de copaíba, cravo-da-índia e manjericão na redução da incidência de fungos associados às sementes de feijão-fava, e sua interferência na qualidade fisiológica. Os tratamentos foram constituídos por óleos essenciais de copaíba, cravo-da-índia e manjericão nas concentrações de 0; 1; 1,5 e 2 mL. L-1 e fungicida (Captan®). Foram utilizadas 200 sementes por tratamento. Para análise da qualidade sanitária das sementes foi adotado o método de incubação em placas de Petri contendo dupla camada de papel-filtro umedecida com ADE, sob temperatura de 20 ±2 ºC, por 7 dias. A qualidade fisiológica das sementes foi determinada com base nos testes de germinação, emergência e vigor das sementes. Para o teste germinação foi empregado método de rolo de papel germitest e na emergência osemeio foi realizado em bandejas plásticas contendo areia lavada esterilizada, em condição de casa de vegetação, e após 9 dias da instalação foi determinado o vigor com base nos resultados de índice de velocidade de germinação e emergência; comprimento da parte aérea, raiz primária e plântula; e teor de massa seca da partes aérea, raízes e plântulas de feijão-fava. O delineamento experimental foi interiamente casualizados, em esquama de fatorial simples (3x4)+1 (óleos essenciais x concentrações + fungicida), com quatro repetições de 50 sementes. Os óleos essenciais de copaíba e manjericão reduziram consideravelmente o percentual de incidência dos fungos associados as sementes de feijão-fava. O óleo essencial de cravo-da-índia na concetração de 2 mL. L-1, reduziu a qualidade fisiológica das sementes de feijão-fava.

ABSTRACT The Phaseolus lunatus L. species, popularly known as lima bean, is one of the alternative sources of income and food for the population of Northeast Brazil. One of the problems is the diseases that cause great losses in seed production. This study evaluated the effectiveness of essential oils of copaiba, clove, and basil in reducing the incidence of fungi associated with lima bean seeds, as well as the effect on their physiological quality. The treatments consisted of essential oils of copaiba, clove, and basil at concentrations of 0; 1; 1.5; and 2 mL. L-1 and fungicide (Captan®). A total of 200 seeds were used per treatment. To analyze the sanitary quality of the seeds, the method used was incubation in petri dishes containing a double layer of filter paper moistened with distilled sterilized water at a temperature of 20 ± 2°C, for seven days. The physiological quality of the seeds was determined by testing their germination, emergence, and vigor. For the germination test, the method used was a roll of Germitest paper. For the emergence, the seeds were sown in plastic trays containing washed sterilized sand in greenhouse conditions. After nine days, the vigor was determined based on the results of germination and emergence speed rates; length of the aerial part, primary root, and seedling; and dry matter content of the aerial part, roots, and seedlings of the lima beans. The experimental design was completely randomized in a simple factorial arrangement (3x4)+1 (essential oils x concentrations + fungicide) with four replications of 50 seeds. The essential oils of copaiba and basil considerably reduced the percentage of incidence of the fungi associated with lima bean seeds. The essential clove oil at 2 mL. L-1 reduced the physiological quality of the lima bean seeds.

Flavonoids and Methoxy-galloylquinic Acid Derivatives from the Leaf Extract of Copaifera langsdorffii Desf.

Despite reports on the pharmacological potential of Copaifera langsdorffii Desf. (Leguminosae-Caesalpinioideae) leaf extract, little is known about its chemical composition. In this work, a phytochemical study from the C. langsdorffii ethanol/H2O 7:3 (v/v) extract was undertaken. Separation was performed by high-speed counter-current (HSCCC) and Sephadex LH-20 column chromatographies, followed by preparative HPLC. The EtOAc- and H2O-soluble fractions of the extract furnished the flavonoids quercitrin (1) and afzelin (2) and 3-O-(3-O-methyl-galloyl)quinic acid (3), respectively. The H2O-soluble fraction furnished 3,4-di-O-(3-O-methyl-galloyl)quinic acid (4), 3,5-di-O-(galloyl)-4-O-(3-O-methyl-galloyl)quinic acid (5), and 3,5-di-O-(3-O-methyl-galloyl)-4-O-(galloyl)quinic acid (6). Their chemical structures were elucidated by NMR means.

Qualidade sanitária e germinação de sementes de copaíba / Sanitary quality and germination of copaiba seeds

Copaifera langsdorffii Desf. pode ser empregada na arborização urbana e na recuperação de áreas degradadas, mas carece de informações sobre análise da qualidade de sementes para a formação de mudas. O objetivo foi determinar a população fúngica associada às sementes, testar métodos para a eliminação desses microrganismos e avaliar a influência desses procedimentos sobre a germinação. Foi realizado o teste de sanidade (método do papel de filtro e do meio BDA), tratamento de sementes (NaClO 2% por 0, 5, 10, 15 e 20 minutos) e o teste de germinação (BOD a 25oC com luz contínua). Os métodos de incubação em papel de filtro e em meio BDA podem ser usados para detecção de fungos em sementes de copaíba, mas o meio BDA é mais eficaz;; o uso de hipoclorito de sódio na concentração de 2% por períodos de 10, 15 ou 20 minutos é capaz de eliminar fungos presentes em sementes de copaíba, entretanto, o período de 15 minutos pode apresentar melhores resultados.

Copaifera langsdorffii Desf. can be used to urban arborization and recomposition of degraded area, but need information about its seed quality to seedlings formation. The aim were determine fungi population associated with seeds, try methods to eliminate it and evaluate the influence of these procedures in seeds germination. Were realized the sanitary test (blotter test and BDA medium),seed treatment (NaClO 2% during 0, 5, 10, 15 and 20 minutes) and germination test (BOD at 25oC with constant light). Blotter test and BDA medium can be used to detect fungi in C. langsdorffi seeds, but BDA is most efficient; NaClO 2% during 10, 15 and 20 minutes is able to eliminate fungi at seeds, but 15 minutes can present best results.

Medicinal plant extracts on the control of Diabrotica speciosa (Coleoptera: Chrysomelidae) / Extratos de plantas medicinais no controle de Diabrotica speciosa (Coleoptera: Chrysomelidae)

The aim of this study was to evaluate the insecticidal effect of aqueous, alcoholic, and oil extracts from leaves of eight medicinal plants against Diabrotica speciosa prepared at five concentrations. The extracts that used commercial soybean oil as solvent showed the highest D. speciosa mortality due to the solvent itself, regardless of the used plants and their concentrations. Thus, commercial soybean oil was discarded as solvent since at these volumes it would cause serious phytotoxicity problems. After 24 hours of exposure of the pest to the extracts, the highest D. speciosa mortality values were observed for Copaifera langsdorfii and Chenopodium ambrosioides extracts, both in 5% alcohol, and Artemisia verlotorum, in 10% water. However, in the last mortality assessment (48 h), C. langsdorfii extract in 5% alcohol showed higher mortality of this pest, followed by C. ambrosioides extract in 5% alcohol, compared to the remaining plants.

O objetivo deste trabalho foi avaliar o efeito inseticida de extratos aquosos, alcoólicos e oleosos de folhas de oito plantas medicinais contra Diabrotica speciosa preparadas em cinco concentrações. Os extratos que utilizaram óleo de soja comercial como solvente apresentaram as maiores mortalidades de D. speciosa em função do próprio óleo, independentemente das plantas utilizadas em suas concentrações. Sendo assim, o óleo de soja comercial foi descartado como solvente, pois nestes volumes acarretaria sérios problemas de fitotoxidade. Após 24 horas de exposição da praga aos extratos, os maiores valores de mortalidade de D. speciosa foram observados nos extratos de Copaifera Langsdorfii e de Chenopodium ambrosioides, ambos em álcool 5%, e de Artemisia verlotorum, em água 10%. Entretanto, na última avaliação de mortalidade (48 h), o extrato de C. langsdorfii em álcool a 5% apresentou maior mortalidade dessa praga, seguida pelo extrato alcoólico a 5% de C. ambrosioides comparada às demais plantas.

Antimicrobial activity of copaíba (Copaifera langsdorffii) oleoresin on bacteria of clinical significance in cutaneous wounds / Atividade antimicrobiana do óleo-resina de copaíba (Copaifera langsdorffii) em bactérias de significância clínica em úlceras cutâneas

The present study was designed to evaluate the in vitro antimicrobial activity of Copaifera langsdorffii oleoresin, which has been used in folk medicine as an anti-inflammatory, antibacterial, healing among others. The oleoresin was tested against Gram-positive (Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacteria related to infections in cutaneous wounds. Antimicrobial activity was determined by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Copaiba oleoresin showed antimicrobial activity only against the Gram-positive bacteria with MIC of 200 µg/mL, 400 µg/mL and 1100 µg/mL for S. aureus, S. pyogenes and E. faecalis, respectively. MBC values were the same as MIC for S. aureus and S. pyogenes and for E. faecalis it was 1200 µg/mL. Considering that infection significantly impairs the wound healing process, we believe that the use of copaiba oleoresin as a component of a topical formulation could be a valuable adjunct in the treatment of infected wounds, mainly in the case of wounds infected by Gram-positive microorganisms.

Este trabalho avaliou a atividade antimicrobiana in vitro do óleo-resina da Copaifera langsdorffii, o qual vem sendo utilizado há muitos anos na medicina tradicional popular, principalmente devido às suas propriedades antiinflamatórias, antibacterianas, cicatrizante entre outras. O óleo-resina foi testado em bactérias Gram-positivas (Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis) e Gram-negativas (Pseudomonas aeruginosa e Escherichia coli) relacionadas com infecções de úlceras cutâneas. A atividade antimicrobiana foi determinada pelos testes da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM). O óleo-resina apresentou atividade antimicrobiana in vitro apenas para as bactérias Gram-positivas, com valores de CIM de 200 µg/mL, 400 µg/mL e 1100 µg/mL para S. aureus, S. pyogenes e E. faecalis, respectivamente. Os valores de CBM foram os mesmos que os valores de MIC para S. aureus e S. pyogenes. O valor de CBM para o microrganismo E. faecalis foi de 1200 µg/mL. Considerando que a presença de infecção significativamente impede o processo normal de cicatrização de úlceras cutâneas, acreditamos que o óleo-resina de copaíba, utilizado como componente de formulações tópicas, poderia ser um adjunto importante no tratamento de úlceras cutâneas infectadas, principalmente nos casos de infecção por microrganismos Gram-positivos.

Parâmetros fisiológicos de mudas de copaíba sob diferentes substratos e condições de sombreamento / Physiological parameters in seedlings of copaiba under different shade conditions and substrates

O presente trabalho teve como objetivo avaliar parâmetros fisiológicos de mudas de copaíba produzidas sob o efeito de diferentes níveis de sombreamento e tipos de substratos. O experimento teve a duração de 130 dias e foi conduzido em blocos casualizados no esquema fatorial 5x4, com cinco substratos: Bioplant®; 70% vermiculita + 30% casca de arroz carbonizada; 40% vermiculita + 30% casca de arroz carbonizada + 30% fibra de coco; 50% vermiculita + 30% casca de arroz carbonizada + 20% areia; 70% vermiculita + 15% casca de arroz carbonizada + 15% vermicomposto de resíduo de indústria têxtil; quatro níveis de sombreamento (100, 70, 50 e 30% do pleno sol) e três repetições. O nível de sombreamento de 50% em relação ao pleno sol possibilitou a produção de mudas de copaíba com maiores teores de clorofila b e clorofila total, além de proporcionar os menores valores de transpiração diária e ao longo do dia das plantas. A área foliar, os teores de clorofila e a transpiração das mudas de copaíba independem do tipo de substrato, podendo ser produzidas em qualquer um dos meios de crescimento avaliados.

This research aimed to evaluate physiological parameters of seedlings of copaiba produced under the effect of different shading levels and types of substrates. The experiment lasted 130 days and was conducted in randomized blocks in factorial scheme 5x4 with five substrates: Bioplant®, 70% vermiculite + 30% rice hulls, 40% vermiculite + 30% rice charred hulls + 30% fiber coconut, 50% vermiculite + 30% rice charred hulls + 20% sand, 70% vermiculite + 15% rice charred hulls + 15% vermicompost residue of textile industry; four shading levels (100, 70, 50 and 30% full sun) and three replications. The shade level of 50% compared to full sun, enabled the production of seedlings of Copaiba with higher concentrations of chlorophyll b and total chlorophyll, and provided the lowest daily transpiration throughout the day and the plants. The leaf area, chlorophyll content and transpiration of seedlings of Copaiba independent of the substrate and can be produced in any of the growth media evaluated.

An efficient and rapid DNA minipreparation procedure suitable for PCR/SSR and RAPD analyses in tropical forest tree species

An efficient and rapid DNA minipreparation modified method for frozen samples was developed for five tropical tree species: Copaifera langsdorffii, Hymenaea courbaril, Eugenia uniflora, Tabebuia roseo alba and Cariniana estrellensis. This procedure that dispenses the use of liquid nitrogen, phenol and the addition of proteinase K, is an adaptation of the CTAB-based DNA extraction method. The modifications included the use of PVP to eliminate the polyphenols, only one chloroform-isoamyl alcohol step and the addition of RNase immediately after extraction with chloroform. The yields of the DNA samples ranged from 25.7 to 42.1 µg from 100 mg leaf tissue. The DNA samples extracted by this method were successfully used for PCR (SSR and RAPD) analyses in these five and other twelve tropical tree species.

Este trabalho teve como objetivo otimizar um protocolo econômico, rápido e eficaz de minipreparação de DNA genômico, para as espécies florestais Copaifera langsdorffii (Óleo de Copaíba), Hymenaea courbaril (Jatobá), Eugenia uniflora (Pitanga), Tabebuia roseo alba (Ipê Branco) e Cariniana estrellensis (Jequitibá Branco). Este método é uma adaptação da técnica de extração CTAB de Doyle e Doyle (1990), o qual consiste principalmente na adição de PVP para eliminar polifenoles, somente uma etapa de extração com clorofórmio-álcool isoamílico e a adição da RNase A imediatamente após a extração com clorofórmio. O método também dispensa o uso de nitrogênio líquido, o uso do fenol e a adição de proteinase K. Os DNAs das espécies florestais extraídos apresentaram alto rendimento e boa qualidade, com rendimento de 25.7 a 42.1 µg de DNA a partir de 100 mg de tecido foliar congelado. Com este protocolo, em apenas 1 dia de trabalho, uma pessoa pode completar o isolamento do DNA de aproximadamente 50 amostras de folhas (dependendo da capacidade da centrífuga). O DNA obtido pode ser usado para métodos de análise baseados em PCR (SSR e RAPD).

Influência do óleo de Copaifera langsdorffii no reparo de ferida cirúrgica em presença de corpo estranho / Influence of Copaifera langsdorffii oil on the repair of a surgical wound in the presence of foreign body

A Copaifera langsdorffii é uma leguminosa nativa do Brasil, da qual pode ser extraído um óleo, popularmente conhecido como óleo de copaíba. Este óleo é amplamente utilizado para tratamento de feridas cutâneas por ser reconhecido como antiinflamatório e cicatrizante. Apesar disso, poucas comprovações científicas do verdadeiro efeito terapêutico desta planta medicinal foram produzidas. O objetivo desse trabalho foi avaliar o efeito do tratamento tópico com C. langsdorffii no processo de reparo quando um corpo estranho é o indutor da inflamação. Para isso, 60 camundongos da linhagem BALB/c foram submetidos à incisão cirúrgica linear de 1cm no dorso para realizar o implante de uma lamínula de vidro com 12mm de diâmetro no tecido subcutâneo. Quatro tratamentos para a ferida foram estabelecidos: controle (C) tratado com solução salina estéril, controle veículo (CV) tratado com óleo mineral estéril, tratamento 1 (T1) tratado com diluição (v:v) de óleo mineral estéril e óleo de C. langsdorffii, tratamento 2 (T2) tratado com óleo de C. langsdorffii puro. As avaliações foram realizadas em períodos de tempo pré-determinados (1, 3, 5, 7 e 14 dias). Era possível perceber características da fase proliferativa como a reepitelização, a presença de fibroblastos e a neovascularização, porém os grupos tratados com o óleo (T1 e T2) não apresentavam reepitelização aos três dias. Esses grupos aos 5 e 7 dias apresentavam no exame macroscópico maior intensidade de edema, hiperemia e permanência de crostas. Na microscopia, a reepitelização ainda não estava completa e a crosta era serocelular. Nos grupos C e CV, apesar de discreta, era predominante a presença de mononucleares, enquanto nos grupos T1 e T2 o infiltrado inflamatório era misto e com maior intensidade que nos outros grupos. Quatorze dias depois da incisão cirúrgica, os aspectos macroscópicos dos grupos C e CV eram semelhantes e os grupos T1 e T2, apesar de a ferida estar completamente fechada e sem...

Copaifera langsdorffii is a Brazilian native leguminosae that produce resin-oil, popularly known as copaíba oil. This oil is used for the treatment of skin wound due to its recognized antiinflammatory and wound healing effects. Despite, its popular use, there are few published data about the therapeutic effect of this medicinal plant. The aim of the study was to evaluate the topic treatment effect of the Copaíba oil on the process of skin repair inflammation induced by a foreign body subcutanously implanted. Sixty BALB/c mice were submitted to a 1cm linear incision and a 12mm circle coverslip was subcutaneously implanted. Four treatments groups were established: control, sterile saline (C); vehicle control, sterile mineral oil, (VC); treatment 1 (T1), mineral oil plus copaiba oil (V/V), and treatment 2 (T2) copaiba oil. The evaluations were performed at pre-determined time points (1, 3, 5, 7 and 14 days). It was possible to find fibroblasts, epithelial cells proliferation, re-epithelization and newly formed blood vessels in all groups, however, all oil treated groups (T1 and T2) did not present re-epithelization at three days post surgical incision. On days 5 and 7, a higher intensity of edema and hyperemia on the groups T1 and T2 was observed, besides that, the T1 and T2 groups presented a serous cellular scab on the wounds that was absent on the C and VC groups. The inflammatory reactions among the groups C and VC showed more mononuclear cells than the T1 and T2 groups that presented a mixed cell patter composed from both mono and polymorphonuclear cells. Although the surgical wounds were re-epithelizaded, in the groups T1 and T2, they were covered by a serous cellular crust and the dermis tissue still presented an intense mononuclear cell inflammatory focus. Fourteen days after of the surgical incision, the gross aspects on groups C and VC were similar and on groups T1 and T2, despite wound to be completely closed and without...