Green manure (Crotalaria juncea L.) enhances Origanum vulgare L. biomass accumulation, essential oil yield, and phytochemical properties.

Green manure (GM) may reduce the use of chemical fertilizers, been an ecologically appropriate strategy to cultivation of medicinal plants. Crotalaria juncea, is one of the most used because it adapts to different climatic and high nitrogen content. Origanum vulgare. is widely used in cooking, pharmaceutical, cosmetic industries and food products. The objectives of this study were to evaluate the GM on biomass, essential oil (EO), phenolic and antioxidant. The experiment consisted: control; 150, 300, 450, and 600 g (Sh= leaves+steam) more 200 g roots (R); 600 g aerial part; 200 g roots; and soil with 300 g cattle manure per pot. The highest dry weights were observed in the presence of GM and cattle manure (90 days). The control had an EO production 75% lower in relation to the dose of 450 g GM (Sh+R). Principal component analysis showed that GM and cattle manure positively influenced the dry weight, content, yield, and EO constituents, and total flavonoids. The GM contributed to the accumulation of the major EO compounds (trans-sabinene hydrate, thymol, terpinen-4-ol). The GM management may be beneficial for cultivating, because it can increase the production of biomass and the active components, in addition to being an inexpensive resource.

Impact of Colored Shade Nets on Biomass Production, Essential Oil Composition and Orientin and Isoorientin Content in Lippia gracilis Schauer.

The objective of this study was to evaluate the effect of ChromatiNet on vegetative growth, total antioxidant capacity, phenolic and essential oils (EOs) composition of Lippia gracilis. The plants were cultivated under full sunlight, black, blue and red ChromatiNet. The flavonoid content and antioxidant capacity were quantified spectrophotometrically. The C-glycosylflavone isomers (orientin and isoorientin) were isolated and identified by conventional spectroscopic techniques and measured using high-performance liquid chromatography-diode array detection. The EO was analysed by gas chromatography and gas chromatography-mass spectrometry. Environment influenced growth, total antioxidant capacity and phytochemical levels. Shoot dry weight, thymol, carvacrol and (E)-caryophyllene were favoured under red and black ChromatiNet. Root growth, EOs, caryophyllene oxide, p-cymene, flavonoids, orientin and isoorientin were favoured in sunlight. Growth and accumulation of EOs, flavonoids and photosynthetic pigments increased under blue ChromatiNet. Therefore, Lippia gracilis plants have plasticity related to the spectral quality of light and it cultivate depends of the phytochemicals of interest.

Inhibition of α-Glycosidase by Lippia dulcis Trevir. (Verbenaceae) Preparations, Quantification of Verbascoside, and Study of Its Molecular Docking.

This study aimed to quantify verbascoside (VEB), perform molecular docking studies of VEB with the α-glucosidase (GL) of Bacillus stearothermophilus, and evaluate the inhibition of the enzyme by L. dulcis preparations. The substrate concentration and presence of reduced glutathione were evaluated for their effect on the in vitro inhibition of the GL enzyme. Assays were also performed in the presence and absence of simulated gastric fluid. The antidiabetic fractions 2 and 3 were the most inhibited GL, but their activity were significantly decreased in the presence of gastric fluid. Chromatographic analyses confirmed the predominant presence of VEB in the samples. The samples had VEB concentrations between 49.9 and 243.5 mg/g. Simulation of the molecular docking of VEB were consistent with its GL-inhibitory activity. It can conclude that the crude ethanol extract and fractions show inhibitory activity against the GL enzyme.

Solvent Mixture Optimization in the Extraction of Bioactive Compounds and Antioxidant Activities from Garlic (Allium sativum L.).

Garlic is a health promoter that has important bioactive compounds. The bioactive extraction is an important step in the analysis of constituents present in plant preparations. The purpose of this study is to optimize the extraction with the best proportion of solvents to obtain total phenolic compounds (TPC) and thiosulfinates (TS) from dried garlic powder, and evaluate the antioxidant activities of the optimized extracts. A statistical mixture simplex axial design was used to evaluate the effect of solvents (water, ethanol, and acetone), as well as mixtures of these solvents, after two ultrasound extraction cycles of 15 min. Results showed that solvent mixtures with a high portion of water and pure water were efficient for TPC and TS recovery through this extraction procedure. According to the regression model computed, the most significant solvent mixtures to obtain high TPC and TS recovery from dried garlic powder are, respectively, the binary mixture with 75% water and 25% acetone and pure water. These optimized extracts presented oxygen radical absorbance capacity. Pure water was better for total antioxidant capacity, and the binary mixture of water-acetone (75:25) was better for DPPH scavenging activity. These optimized extracts can be used for industrial and research applications.

In Vitro Anticoagulant Activity of Mikania laevigata: Deepening the Study of the Possible Interaction Between Guaco and Anticoagulants.

Mikania laevigata, popularly known in Brazil as guaco, is widely used for respiratory disorders. As this plant is rich in coumarins, there is evidence of indications that it may cause bleeding and therefore should not be used concomitantly with anticoagulants. The basis of this information is very theoretical, with no clinical evidence of such contraindication. Thus, the aim of this study was to evaluate the in vitro effect of M. laevigata extract on blood coagulation through prothrombin time (PT) and activated partial thromboplastin time (aPTT) tests, fibrinogen plasma concentration, and the new thrombin generation test, which investigate, with high sensibility, hemostatic changes (CAAE 60904316.6.0000.5149), besides evaluating its qualitative micromolecular composition, providing scientific evidence to support the management of patients taking warfarin. Ethanolic extracts of guaco leaves were incubated with a plasma pool of healthy individuals at concentrations of 1.67, 2.26, and 2.86 mg/mL. The presence of flavonoids, tannins, coumarins, and triterpenes was demonstrated by selective reagents in thin layer chromatography. Benzoylgrandifloric acid, cinnamoylgrandifloric acid, o-coumaric acid, coumarin, and quercetin-3-ß-glucoside were identified by coinjection in ultraperformance liquid chromatography. The extract at all concentrations prolonged TP and aPPT and reduced the potential for endogenous thrombin potential by the thrombin generation test. The control plasma had endogenous thrombin potential = 1465 nM/min, and after the addition of M. laevigata extract (2.26 mg/mL), this value was reduced to 1087 nM/min, indicating a lower generation of thrombin. Related to fibrinogen plasma concentration, concentrations of 2.26 and 2.86 mg/mL were effective in reducing plasma fibrinogen levels. These results allow us to conclude that the guaco extract demonstrated an anticoagulant effect in vitro, possibly interfering with intrinsic, extrinsic, and common coagulation pathways. A discussion on the contribution of the identified substances to the activity is also present.

Growth and production of volatile compounds of yarrow (Achillea millefolium L.) under different irrigation depths.

Yarrow (Achillea millefolium L., Asteraceae) is an important medicinal plant used worldwide for its medicinal properties such as the analgesic, antioxidant and anti-inflammatory ones. The aim of this study was to evaluate the growth and production of photosynthetic pigments and of volatile constituents of Achillea millefolium L. under different irrigation depths. The treatments were the application of 55, 110, 220, 440 and 880 mm of water for a period of 110 days. Data were submitted to polynomial regression analysis at 5% probability, while the volatile constituents were analyzed by standard deviation. Different irrigation depths provided quadratic growth responses being the highest dry matter production at the depth of 440 mm. The contents of chlorophyll a, b, total and carotenoids were higher at the lower depth tested (55 mm). The major volatile compounds identified were sabinene, 1,8-cineol, borneol and ß-caryophyllene. Increased water availability reduced the complexity of the volatile fraction of essential oil. Thus, it is recommended that the species be cultivated at 440 mm irrigation depth to have a higher production of dry matter and lower variation in the volatile profile of the essential oil.

Colored shade nets induced changes in growth, anatomy and essential oil of Pogostemon cablin.

The purpose of this investigation was to determine the influence of colored shade nets on the growth, anatomy and essential oil content, yield and chemical composition of Pogostemon cablin. The plants were cultivated under full sunlight, black, blue and red nets. The harvesting was performed 5 months after planting and it was followed by the analysis of plant growth parameters, leaf anatomy, essential oil content, yield and chemical composition. The plants grown under red net have produced more leaf, shoot, total dry weight and leaf area. Plants cultivated under colored nets showed differences in morphological features. Plants maintained under red net had a higher leaf blade thickness and polar and equatorial diameter of the stomata ratio. Additionally, higher yield of essential oil in the leaves was observed under red and blue colored shade net. The essential oil of the plants grown under red net showed the highest relative percentage of patchoulol (66.84%). Therefore, it is possible using colored shade nets to manipulate P. cablin growth, as well as its essential oil production with several chemical compositions. The analyses of principal components allowed observing that pogostol has negative correlation with α-guaiene and α-bulnesene. There was difference in total dry weight and patchoulol content when the patchouli is cultured under the red colored shade nets.

Effects of Essential Oils on Biological Characteristics and Potential Molecular Targets in Spodoptera frugiperda.

Spodoptera frugiperda control methods have proved to be inefficient, which justifies the search for new control measures. In this search for botanical insecticides for controlling S. frugiperda, the following were evaluated: (i) the toxicity of essential oils (EOs) from Cinnamodendron dinisii, Eugenia uniflora, and Melaleuca armillaris; (ii) the effect of EOs on life table parameters against S. frugiperda; (iii) the chemical characterization of EOs; and (iv) the in silico interaction of the chemical constituents present in the three EOs with the molecular targets of S. frugiperda. The EO from E. uniflora had the lowest LD50 (1.19 µg of EO/caterpillar). The major compounds bicyclogermacrene (18.64%) in C. dinisii and terpinolene (57.75%) in M. armillaris are highly predicted to interact with the octopamine receptor (OctpR). The compound 1,8-cineole (21.81%) in M. armillaris interacts mainly with a tolerant methoprene receptor (MET) and curzerene (41.22%) in E. uniflora, which acts on the OctpR receptor. Minor compounds, such as nerolidol in C. dinisii and ß-elemene in E. uniflora, are highly ranked for multiple targets: AChE, MET, OctpR, and 5-HT1. It was concluded that the EO from E. uniflora negatively affects several biological parameters of S. frugiperda development and is promising as an active ingredient in formulations for controlling this insect pest.

Evaluation of the Anti-Inflammatory and Antioxidant Potential of Cymbopogon citratus Essential Oil in Zebrafish.

This study explored the protective capacity of the essential oil (EO) of Cymbopogon citratus against oxidative stress induced by hydrogen peroxide (H2O2) and the inflammatory potential in zebrafish. Using five concentrations of EO (0.39, 0.78, 1.56, 3.12, and 6.25 µg/mL) in the presence of 7.5 mM H2O2, we analyzed the effects on neutrophil migration, caudal fin regeneration, cellular apoptosis, production of reactive oxygen species (ROS), and activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) after 96 h of exposure. A significant decrease in neutrophil migration was observed in all EO treatments compared to the control. Higher concentrations of EO (3.12 and 6.25 µg/mL) resulted in a significant decrease in caudal fin regeneration compared to the control. SOD activity was reduced at all EO concentrations, CAT activity significantly decreased at 3.12 µg/mL, and GST activity increased at 0.78 µg/mL and 1.56 µg/mL, compared to the control group. No significant changes in ROS production were detected. A reduction in cellular apoptosis was evident at all EO concentrations, suggesting that C. citratus EO exhibits anti-inflammatory properties, influences regenerative processes, and protects against oxidative stress and apoptosis.

Isolation and HPLC quantitation of kaurane-type diterpenes and cinnamic acid derivatives of long-term stored leaves of Mikania laevigata and Mikania glomerata.

The leaves of Mikania laevigata and Mikania glomerata are used in Brazil to treat respiratory affections, being kaurane-type diterpenes and coumarin considered as the bioactive compounds. The present study reports an investigation on the HPLC-DAD profiles and contents of coumarin (1), trans-o-coumaric (2), kaurenoic (3), benzoylgrandifloric (4) and cinnamoylgrandifloric (5) acids in dried leaves of Mikania species stored in dark room under controlled conditions. Excepting 2, the constituents were isolated and purified to be employed as reference compounds. The samples were analyzed at three monthly intervals up to 18 months for M. laevigata and 12 months for M. glomerata. trans-o-Coumaric was not detected in both, whereas 1 occurred only in M. laevigata. The concentrations of the assayed constituents did not vary significantly within the evaluated period (p < 0.05), for both species. In contrast, changes in the chromatographic profiles and spectral purity of peaks from 3, 4 and 5 were detected in samples of both Mikania stored for three months, while the coumarin profile in M. laevigata modified after six months of storage. The evaluation of chromatographic profiles based on spectral purity analyses of selected peaks was shown to be a more robust tool to access chemical stability of Mikania samples than the quantitation of chemical markers' contents.

Seasonal variation on the contents of coumarin and kaurane-type diterpenes in Mikania laevigata and M. glomerata leaves under different shade levels.

Coumarin (1) and kaurane-type diterpenes are considered the bioactive constituents of Mikania glomerata and M. laevigata, used in Brazil to treat respiratory affective disorders. The seasonal variation of 1, ortho-coumaric acid (2), benzoylgrandifloric acid (3), cinnamoylgrandifloric acid (4), and kaurenoic acid (5) in leaves of both species, cultivated in full sunlight and under shade levels of 40 and 80%, was quantified by HPLC. Compound 2 was detected solely in M. laevigata in concentrations below the limit of quantification. Coumarin was not found in M. glomerata, whereas its concentration reached 0.94±0.24% (w/w) in M. laevigata farmed in summer under 80% shading. Both Mikania species produced higher amounts of kaurane diterpenes when cultivated in plenty of sunlight. Hence, maximum contents of 1 are reached in M. laevigata cultivated under high shading, but with reduced concentrations of 3-5. Conversely, M. glomerata should be cultivated under full sunlight and harvested in winter for highest concentrations of kaurane-type diterpenes.

Elucidating the molecular mechanisms of essential oils' insecticidal action using a novel cheminformatics protocol.

Essential oils (EOs) are a promising source for novel environmentally safe insecticides. However, the structural diversity of their compounds poses challenges to accurately elucidate their biological mechanisms of action. We present a new chemoinformatics methodology aimed at predicting the impact of essential oil (EO) compounds on the molecular targets of commercial insecticides. Our approach merges virtual screening, chemoinformatics, and machine learning to identify custom signatures and reference molecule clusters. By assigning a molecule to a cluster, we can determine its most likely interaction targets. Our findings reveal that the main targets of EOs are juvenile hormone-specific proteins (JHBP and MET) and octopamine receptor agonists (OctpRago). Three of the twenty clusters show strong similarities to the juvenile hormone, steroids, and biogenic amines. For instance, the methodology successfully identified E-Nerolidol, for which literature points indications of disrupting insect metamorphosis and neurochemistry, as a potential insecticide in these pathways. We validated the predictions through experimental bioassays, observing symptoms in blowflies that were consistent with the computational results. This new approach sheds a higher light on the ways of action of EO compounds in nature and biotechnology. It also opens new possibilities for understanding how molecules can interfere with biological systems and has broad implications for areas such as drug design.

Eugenia uniflora, Melaleuca armillaris, and Schinus molle essential oils to manage larvae of the filarial vector Culex quinquefasciatus (Diptera: Culicidae).

Populations of Culex quinquefasciatus Say, 1823 (Diptera: Culicidae) have shown resistance to insecticides of the carbamate and organophosphate classes. The objective of this study was to assess the susceptibility of C. quinquefasciatus larvae to essential oils from leaves of Eugenia uniflora L., Melaleuca armillaris (Sol. ex Gaertn.) Sm., and Schinus molle L and C. quinquefasciatus larvae's biochemical responses after their exposure to these leaves. The essential oils were chemically analyzed by GC and GC/MS. First, the lethal concentration for 50% (LC50) values was estimated using different concentrations of essential oils and probit analysis. The larvae were exposed for 1 h at the LC50 estimated for each essential oil. The susceptibility of the larvae to essential oils was evaluated using the following biochemical parameters: concentrations of total protein and reduced glutathione; levels of production of hydrogen peroxide and lipid peroxidation; and the activity of the enzyme acetylcholinesterase (AChE). The main chemical constituents in E. uniflora were E-ß-ocimene, curzerene, germacrene B, and germacrone; in M. armillaris were 1,8-cineole and terpinolene; and in S. molle were sabinene, myrcene, and sylvestrene. The essential oils had LC50 values between 31.52 and 60.08 mg/L, all of which were considered effective. All of them also promoted changes in biochemical parameters when compared to the control treatment. The essential oils of S. molle and E. uniflora inhibited the activity of the AChE enzyme, and the essential oil of M. armillaris increased it. All essential oils had larvicidal activity against C. quinquefasciatus, but the essential oil of E. uniflora was the most efficient. Thus, the findings of the present study suggest that the essential oil of E. uniflora can be considered promising for the development of botanical larvicides.

In vitro anticoagulant activity of selected medicinal plants: potential interactions with warfarin and development of new anticoagulants.

OBJECTIVES: Warfarin is the most widely used anticoagulant in the world, but it has several limitations including its narrow therapeutic range, need for dose adjustment and high potential for interactions. The simultaneous use of other drugs or even medicinal plants and certain foods could interfere with its therapeutic activity. In this context, this study aims to investigate the in vitro anticoagulant potential and phytochemical constitution of 17 plants selected from a previous clinical cross-sectional study (2014), that investigated the habits of plant utilization among patients taking warfarin. METHODS: Ethanol extracts and essential oils were evaluated, in vitro, as to their effect in the prothrombin time (PT) and activated partial thromboplastin time (aPTT) tests. Four species that presented aPTT >50 s were selected for phytochemical evaluation. RESULTS: Thirteen of the 17 plants selected demonstrated a significant anticoagulant effect in at least one of the evaluated parameters. Citrus sinensis (PT=14.75 and aPTT=53.15), Mentha crispa (aPTT=51.25), Mikania laevigata (PT=14.90 and aPTT=52.10), and Nasturtium officinale (aPTT=50.55) showed greater anticoagulant potential compared to normal plasma pool (PT=12.25 and aPTT=37.73). Chemical profiles of these four species were obtained, and certain compounds were identified: rosmarinic acid from M. crispa and isoorientin from N. officinale. CONCLUSIONS: Thus, the results of this study could be a useful indicator for clinical practice towards the possibility of interaction between these plants and anticoagulants, although further clinical research is needed taking into consideration the limitations of in vitro studies. These findings also suggest that further research into the action of these plants could be of real clinical value in identifying potential alternative anticoagulant therapies.

Biocontrol potential of methyl chavicol for managing Spodoptera frugiperda (Lepidoptera: Noctuidae), an important corn pest.

Synthetic insecticides applied to control Spodoptera frugiperda (Lepidoptera: Noctuidae) can have negative impacts on environment and human health. Botanical essential oils can be sources of organic molecules with biocontrol potential and advantages, such as minor impacts on the selection of resistant pest insects and low toxicity to humans. The aim of this study was to investigate the biocontrol action of essential oils from Brazilian species and methyl chavicol compounds on the development and metabolism of S. frugiperda. Essential oils of Eremanthus erythropappus (Asteraceae), Ocimum selloi, Hyptis suaveolens, and Hyptis marrubioides (Lamiaceae) were distilled by the steam distillation method and analyzed by gas chromatograph techniques. The essential oils were incorporated into an artificial diet (at 1, 2, and 4 mg mL-1) and offered to S. frugiperda caterpillars. Larvae of S. frugiperda at 48 h of age were fed an artificial diet containing the major constituent of O. selloi (methyl chavicol). The major compounds of the essential oils were methyl chavicol for O. selloi, α-bisabolol for E. erythropappus, bicyclogermacrene for H. suaveolens, and ß-thujone for H. marrubioides. O. selloi caused 100% mortality in S. frugiperda larvae at a concentration of 1 mg mL-1 after 48 h. H. marrubioides essential oil caused 100% mortality in larvae at a concentration of 4 mg mL-1 after 48 h. O. selloi and H. marrubioides inhibited acetylcholinesterase (AchE) activity in 72.87% and 81.69% of larvae, respectively. O. selloi presented the highest toxicity to S. frugiperda and the lowest inhibition of AchE. Methyl chavicol was lethal to all larvae within 24 h at a concentration of 0.92 mg mL-1 of diet. Methyl chavicol showed the best insecticidal activity and potential to be used as a natural insecticide to control S. frugiperda.

Chemical composition and anti-inflammatory activity of essential oil and ethanolic extract of Campomanesia phaea (O. Berg.) Landrum leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Campomanesia species are used in folk medicine for anti-inflammatory, -ulcerogenic, -diabetic, -obesity, and many other purposes. AIM OF THE STUDY: This study aimed to investigate the phytochemical profile and pharmacotherapeutic potential of the essential oil (EO) and ethanolic extract (EXT) of the leaves of Campomanesia phaea in relation to antioxidant and anti-inflammatory effects using chemical methods and in vitro bioassays in cell culture. MATERIALS AND METHODS: Gas and liquid chromatography techniques coupled to mass spectrometry were used to identify the main secondary metabolites. The antioxidant activity was determined by the chemical methods of radical sequestration of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and by ferric reducing antioxidant power (FRAP); in addition to the protective effect against cellular oxidative damage caused by hydrogen peroxide (H2O2) in macrophage culture. The anti-inflammatory and immunomodulatory activity was evaluated for the influence on the production of nitric oxide and superoxide anion (O2•-), and by the quantification of proinflammatory cytokines tumor necrosis factor (TNF alpha) and interleukin 6 (IL- 6) through Enzyme Linked Immuno Sorbent Assay (ELISA) technique and inhibition of nuclear factor kappa B (NF-κB) through chemiluminescence. RESULTS: A total of 41 compounds were identified in the essential oil (EO), being (E)-caryophyllene (14%) and caryophyllene oxide (6.9%) the major compounds. In the ethanolic extract (EXT), three flavonoids from the flavanones group were identified: alpinetin O-dideoxy-hexoside, 5,7-dimethoxyflavanone and alpinetin. The EO and EXT inhibited the production of O2•- (99.0% and 52.9%) at a concentration of 100 µg/mL, intracellular NO•- (50.0% and 51.9%) and proinflammatory cytokines IL-6 (41.0% and 82.9%) and TNF-α (74.7% and 87.9%) at a concentration of 50 µg/mL, respectively. In addition, inhibition of nuclear factor kappa B (EO 36.2% and EXT 40.9%) was observed at 20 µg/mL. CONCLUSIONS: Taken together, the results indicated that EO and EXT possess potent anti-inflammatory activities and it may hold therapeutic promise in the management of acute and chronic inflammatory conditions.

Study of the influence of wavelengths and intensities of LEDs on the growth, photosynthetic pigment, and volatile compounds production of Lippia rotundifolia Cham in vitro.

Lippia rotundifolia Cham. is in the family Verbenaceae and is endemic to the Cerrado. This species is aromatic and characterized by the presence of glandular trichomes on its leaves that are rich in monoterpenes. The objective of this study was to evaluate the growth, photosynthetic pigment production, and chemical composition of L. rotundifolia grown in vitro under different light wavelengths and intensities. The light intensities consisted of five treatments using cool white fluorescent lamps at 20, 54, 78, 88, and 110 µmol m-2 s-1. The light quality consisted of six treatments using light-emitting diodes (LEDs) in different light wavelengths, namely, white, red, blue, and their interactions: 1R:1B, 2.5R:1B, and 1R:2.5B. After 45 days, the biometric parameters, photosynthetic pigment content, and volatile compounds were evaluated. The lower light intensities of 20 and 54 µmol m-2 s-1 generated higher growth, photosynthetic pigment content, and biomass accumulation. Myrcene and pentadecane were highest under light intensities of 88 and 110 µmol m-2 s-1, respectively. The highest limonene and ocimenone levels were obtained at 20 and 54 µmol m-2 s-1 intensity, respectively, and the highest myrcenone content was obtained at 78 µmol m-2 s-1 intensity. Regarding the light wavelengths, the combination of red and blue spectra further stimulated plantlet growth, and the 2.5R:1B combination obtained the best biometric data and total chlorophyll content. The z-ocimenone chemical compound contents were highest under the 1R:2.5B light spectrum. The monochromatic blue spectrum increased the myrcene and limonene content but decreased the myrcenone content, which was increased by red light. The highest pentadecane contents were obtained with the white spectrum and the red and blue combinations.

Biological screening of herbal extracts and essential oil from Plectranthus species: α-amylase and 5-lipoxygenase inhibition and antioxidant and anti-Candida potentials

Abstract The phenolic compound content, the antioxidant and α-amylase inhibition potentials of different extracts of the Plectranthus amboinicus, P. barbatus and P. ornatus were evaluated. We also evaluated the influence of plant growth and harvest time on the chemical composition of the essential oil (EO) of P. amboinicus, its antioxidant and anti-Candida activities and the α-amylase and lipoxygenase inhibitions. The turbo-extract of P. barbatus showed the greatest phenolic compound content and antioxidant activity. No α-amylase inhibition activity was observed in the analyzed extracts, but the turbo-extraction and refluxing extracts possessed high antioxidant activities. Protected cultivation and morning harvest conditions gave the best antioxidant activities, which was associated to the highest carvacrol content. P. amboinicus EO antioxidant activity could contribute to the reduction of oxidative stress in diabetes. Causal Candida strains of diabetic foot ulcers showed sensitivity to P. amboinicus EO. C. albicans and C. dubliniensis were the most sensitive of the selected Candida strains. Turbo-extracts or refluxing of the three species extracts and the EO of P. amboinicus should be considered as a potential candidate for the management the complications of type 2 diabetes.

Seasonal variation in essential oil content and chemical profile of mint in southeast of Brazil / Variação sazonal no teor e perfil químico do óleo essencial de menta no sudeste do Brasil

ABSTRACT: This study evaluated the influence of seasonality in essential oil (EO) chemical composition of Mentha x piperita L. and Mentha viridis L. grown in southeast of Brazil. Leaves were collected from November 2011 until August 2013, in the middle of the following seasons: Spring (November), Summer (February), Autumn (May) and Winter (August). EO extraction was carried out by hydrodistillation in Clevenger apparatus for 1 hour. Chemical composition of EO was analyzed by GC-FID and GC-MS. The EO content and its chemical composition were influenced by seasonality in both species. The highest EO content was obtained for M. x piperita during spring (4.26%) and for M. viridis during spring (3.30%) and summer (3.70%). Mentha x piperita increased menthol (16.31 to 41.26%), neomenthol (3.02 to 6.39%) and menthone (5.56 to 41.58%) contents during spring and summer, respectively. Whereas, M. viridis EO content did not show a quality response pattern in relation to seasonality. Therefore, harvest time for M. x piperita is recommended during spring, and for M. viridis is recommended during spring and summer.

RESUMO: O estudo teve como objetivo avaliar a influência da sazonalidade na composição química do óleo essencial (OE) de Mentha x piperita L. e Mentha viridis L. cultivadas no sudeste do Brasil. As folhas foram coletadas de novembro de 2011 até agosto de 2013, em meados das seguintes estações: primavera (novembro), verão (fevereiro), outono (maio) e inverno (agosto). A extração do OE foi realizada por hidrodestilação em aparelho de Clevenger por uma hora. A composição química do EO foi analisada por GC-FID and GC-MS. O teor de OE e sua composição química foram influenciados pela sazonalidade em ambas as espécies. O maior teor de OE foi obtido para M. x piperita na primavera (4,26%) e para M. viridis na primavera (3,30%) e verão (3,70%). Mentha x piperita aumentou o conteúdo de mentol (16,31 a 41,26%), neomentol (3,02 a 6,39%) e mentona (5,56 a 41,58%) durante a primavera e o verão, respectivamente. Já o conteúdo de OE de M. viridis não apresentou padrão de resposta de qualidade em relação à sazonalidade. Portanto, a colheita para M. x piperita é recomendada durante a primavera, e para M. viridis é recomendada durante a primavera e verão.

Sperm Quality and Testicular Histomorphometry of Wistar Rats Supplemented with Extract and Fractions of Fruit of Tribulus terrestris L.

ABSTRACT The aim of this study was to assess the sperm quality and testicular histomorphometry of Wistar rats supplemented with extract and fractions of fruits of Tribulus terrestris L. The ethanolic extract was obtained by dynamic maceration of spray-dried fruit. This extract was fractionated by liquid-liquid partition, using increasing polarity solvents. Twenty male rats were separated in four groups, with five rats in each group. The control was supplemented with distilled water, while the others were daily given the ethanolic extract, hexanic or aqueous fraction soluble in methanol in a dose of 42 mg.kg-1.day-1 for 70 days. Sperm was obtained from the right epididymal tail for the analysis of motility, count, morphology and viability. The testicular weight of groups supplemented with ethanolic extract and aqueous fraction soluble in methanol was higher when compared to the control. The gonadosomatic index increased in the group supplemented with ethanolic extract. The nuclear, cytoplasmic and individual volume of Leydig cells increased in supplementation with hexanic and aqueous fractions soluble in methanol. It was concluded that the extract influenced the spermatogenesis, while hexanic and aqueous fractions soluble in methanol promoted the changes in the intertubular compartment. Therefore, Tribulus terrestris did not improve the sperm quality of the rats.