Effect of ethanolic extracts from Piperaceae leaves on the reduction of skin necrosis and wound healing in an animal model of degloving injuries

Purpose: To assess the effect of the ethanolic extract of the leaves of three species of plants from the Piperaceae family on reducing necrosis and enhancing wound healing in an animal model of degloving injuries. Methods: The animals were divided into six groups, each consisting of six animals: sham, negative control, EEPA (Piper amalago ethanolic extract), EEPG (Piper glabratum ethanolic extract), EEPV (Piper vicosanum ethanolic extract), and positive control receiving hyperbaric oxygenation. The animals underwent surgery to induce excision wounds, and the extent of cutaneous necrosis was evaluated using graphic software, while wound healing was assessed through histopathology. Results: Skin necrosis percentage area was: sham group = 62.84% 6.38; negative control group = 63.03% 4.11; P. vicosanum = 40.80% 4.76 p < 0.05; P. glabratum 32.97% 4.01 p < 0.01; P. amalago = 32.40% 4.61 p < 0.01; hyperbaric oxygenation = 33.21% 4.29 p < 0.01. All treated groups showed higher collagen deposition and less intense, plus predominantly mononuclear inflammatory infiltrate, suggesting improved healing process. Conclusions: The three tested extracts demonstrated efficacy in reducing the extent of cutaneous necrosis caused by degloving injuries and also showed evidence of improvement in the wound healing process.

Anti-acanthamoebic activity of methanolic extract of Piper sarmentosum leaves

Aims@#Piper sarmentosum or locally known as Kaduk, is a tropical herb plant that was investigated for its phenolic content by previous researchers. The present study aimed at the analysis of crude methanolic extract of P. sarmentosum leaves for phenolic compounds identification and its anti-amoebic properties against pathogenic Acanthamoeba castellanii.@*Methodology and results@#Folin-Ciocalteu assay was used to determine P. sarmentosum leaves methanolic extract (PSLME)’s total phenolic content (TPC). The extract was further characterized by using gas chromatography-mass spectrometry (GC-MS), reverse phase-high performance liquid chromatography (RP-HPLC) and liquid chromatography-mass spectrometry (LC-MS) analyses to determine the chemical constituents in methanolic PSLME extract. The cytotoxicity of the extract was evaluated through the determination of inhibition concentration for half of cell population (IC50) of pathogenic A. castellanii followed by cell morphological analysis using inverted light and scanning electron microscopies. Acridine-orange/Propidium iodide (AOPI) staining was also conducted to determine the integrity of cell membrane for quantitative analysis. The results demonstrated that the TPC from PSLME was 142.72 mg [GAE]/g with a total of 33 phenolic compounds identified. The IC50 value obtained for A. castellanii was low (74.64 μg/mL) which indicates promising anti-acanthamoebic activity. Microscopy analyses showed that the plant extract caused cells encystment, in which exhibited by distinctive morphological changes on the cells shape and organelle, as well as shortening of acanthopodia. The dual staining and its quantitative analysis prove compromised membrane integrity in the treated amoeba.@*Conclusion, significance and impact of study@#This finding provides the evidence that PSLME contains active phenolic compounds contributing to the anti-acanthamoebic activity on pathogenic Acanthamoeba species.

Phytochemistry and anti-inflammatory activities of Piper kadsura (Choisy) Ohwi ­ a review / Actividades fitoquímicas y antiinflamatorias de Piper kadsura (Choisy) Ohwi - una revisión

Piper kadsura (Choisy) Ohwi which belongs to the family Piperaceae, is a well-known medicinal plant possessing high medicinal and various therapeutic properties. It is widely used in traditional Chinese medicine for the treatment of asthma and rheumatic arthritis. Numerous studies on this species have also corroborated the significant anti-inflammatory potential of its extracts and secondary metabolites. The main chemical constituents which have been isolated and identified from P. kadsura are lignans and neolignans, which possess anti-inflammatory activities. The present article aims to provide a review of the studies done on the phytochemistry and antiinflammatory activities of P. kadsura. The scientific journals for this brief literature review were from electronic sources, such as Science Direct, PubMed, Google Scholar, Scopus, and Web of Science. This review is expected to draw the attention of the medical professionals and the general public towards P. kadsura and to open the door for detailed research in the future.

Piper kadsura (Choisy) Ohwi, perteneciente a la familia Piperaceae, es una planta medicinal conocida que posee importantes propiedades medicinales y diversas propiedades terapéuticas. Es ampliamente utilizada en la medicina tradicional china para el tratamiento del asma y la artritis reumática. Numerosos estudios sobre esta especie también han corroborado el destacado potencial antiinflamatorio de sus extractos y metabolitos secundarios. Los principales componentes químicos que se han aislado e identificado de P. kadsura son los lignanos y los neolignanos, que poseen actividades antiinflamatorias. El presente artículo tiene como objetivo proporcionar una revisión de los estudios realizados sobre las actividades fitoquímicas y antiinflamatorias de P. kadsura. Las revistas científicas para esta breve revisión de literatura fueron de fuentes electrónicas, como Science Direct, PubMed, Google Scholar, Scopus y Web of Science. Se espera que esta revisión atraiga la atención de los profesionales médicos y el público en general respecto de P. kadsura y abra la puerta a una investigación detallada en el futuro.

Actividad in vitro anti-Leishmania (Trypanosomatidae) del epóxido trans-Z-&-bisaboleno y del Safrol, en frutos de Piper auritum (Piperaceae) / Anti-Leishmania (Trypanosomatidae) activity of trans-Z-&-bisabolene epoxide and isolation of safrole, in fruits of Piper auritum (Piperaceae)

Resumen La leishmaniasis es una enfermedad con una alta incidencia en el ser humano, que puede ser controlada, pero como los tratamientos tienen efectos secundarios importantes se han realizado estudios de diversas plantas con el fin de encontrar compuestos con actividad antileishmaniásica que presenten pocos efectos nocivos para el ser humano. El presente estudio consistió en realizar un tamizaje fitoquímico de la planta, para identificar la presencia de cumarinas, terpenos, triterpenos y azúcares reductores. El objetivo fue encontrar componentes químicos puros con actividad contra el parásito Leishmania sp. Por tal razón se purificaron los compuestos: trans-Z-alfabisaboleno y el Safrol, a los que se les realizaron pruebas del efecto anti parasitario que presentaron un CI50 de 50.0 µg/mL y 0.0 µg/mL, respectivamente. Además, se discute la importancia de estos nuevos hallazgos. El compuesto mayoritario presente en los aceites esenciales (Safrol) no es el componente que presentó la actividad. Es importante realizar estudios sobre su proyección en el tratamiento de la leishmaniasis.

Abstract The leishmaniosis disease incidence is high in tropical regions, and its current treatment has shown severe secondary effects. Considering this problem, many studies have focused on plants, looking for chemical components that have anti-leishmanial activity, and are free of adverse effects for human beings. The purpose of this work was to find a chemical component with this kind of activity in Piper auritum. In a phytochemical screening of this plant, we found some cumarins, terpens, triterpens and reducing sugars; and later, we identified the components trans-Z-α-bisabolene epoxide and Safrol. The first component presented a CI50 of 50.0 µg/mL of anti-Leishmania activity. The Safrol, which is the major component of the essential oils of this plant, did not show antiparasitic activity. These results are discussed considering treatment of leishmaniasis. Rev. Biol. Trop. 66(2): 826-835. Epub 2018 June 01.

First phytochemical description of essential oils from Piper cachimboense (Piperales, Piperaceae) / Primeira descrição fitoquímica de óleos essenciais de Piper cachimboense (Piperales, Piperaceae)

Piper cachimboense is recorded only for the Amazon region of Brazil and Colombia, and the objective of this study was to report the first phytochemical assessment of the composition of the essential oils (EOs) from this species collected in the Amazon rainforest, in Novo Progresso, Pará State, Brazil. Samples of leaves were subjected to hydrodistillation in a Clevenger-type apparatus. The chemical identification was carried out by gas chromatography. The yield of oils was of 11.03 ± 5.94% for fresh leaves, and 1.07 ± 0.27% for dry leaves. The analysis showed 36 volatile compounds from fresh leaves and 49 from dried leaves. Main constituents in EOs of both fresh and dried leaves from P. cachimboense were (E)-caryophyllene, germacrene-D, -amorfene, -cadinene and apiole.

Piper cachimboense é registrada apenas para a região amazônica do Brasil e Colômbia, e o objetivo deste estudo foi relatar, pela primeira vez, a composição fitoquímica dos óleos essenciais (OEs) desta espécie coletada na floresta amazônica, em Novo Progresso/PA, Brasil. Amostras de folhas foram submetidas a hidrodestilação em aparelho tipo Clevenger. A identificação química foi realizada por cromatografia gasosa e o rendimento dos óleos foi de 11,03 ± 5,94% para as folhas frescas e de 1,07 ± 0,27% para as folhas secas. A análise mostrou 36 compostos voláteis para folhas frescas e 49 para folhas secas. Os constituintes principais dos OEs de folhas frescas e secas de P. cachimboense foram (E)-cariofileno, germacreno-D, -amorfeno, -cadineno e apiol.

4-Nerolidylcatechol induces autophagy in human glioblastoma cells

ABSTRACT Gliomas account for the majority of primary malignant brain tumors and present invasive behavior into adjacent healthy tissue. While 4-NC had previously shown to induce apoptotic cell death in a melanoma model, for the glioma model described in this paper 4-NC is cytotoxic for the cells with the induction of the autophagic pathway. Trypan blue exclusion assay showed that 4-NC was cytotoxic in a dose-dependent manner for A172 and T98G cell lines. IC10 and IC50 values were at 32 µM and 41 µM for A172 and T98G respectively. Inhibition of cell proliferation was observed by total cell counts and by cell cycle analysis by flow cytometry, with cell cycle arrest of A172 and T98G cell lines respectively in the G1/G0 and S phases of the cell cycle. 4-NC induced up-regulation of autophagic pathways, as shown by immunoblotting for LC3-I/II, Real-Time PCR for ATG-7 and Beclin-1 genes, and by fluorescence microscopy observation of autophagic vacuoles in cells transfected with GFP-LC3 and electron microscopy. Glioma cells concomitantly treated with 4-NC and 3-MA, an inhibitor of the autophagic process, are more sensible to cell death, suggesting that autophagy protects the cells from the action of 4-NC.

Grandisin induces apoptosis in leukemic K562 cells

Abstract In this study, the potential antileukemic activity of grandisin, a lignan extracted from Piper solmsianum, was evaluated against the leukemic line K562. The cytotoxicity of grandisin (0.018 to 2.365 µM) was evaluated in K562 and normal peripheral blood lymphocytes by Trypan Blue Exclusion and MTT methods after 48h exposure to the drug. In both methods, cellular viability was concentration-dependent and the IC50 values were lower than 0.85µM. Analysis of K562 cells after treatment with grandisin showed that the cell cycle was arrested in the G1 phase with a 12.31% increase, while both S and G2 phases decreased. Morphological studies conducted after the exposure of K562 to grandisin revealed changes consistent with the apoptosis process, which was confirmed by anexin V stain and caspase activation. Thus, lignan grandisin showed antileukemic activities against the K562 cell line and the cell death process occurred via apoptosis.

Caracterización fitoquímica y evaluación de actividad inhibitoria sobre acetilcolinesterasa de hojas de Piper pesaresanum C. DC / Phytochemical characterization and evaluation of the inhibitory activity of Piper pesaresanum C. DC leaves against acetylcholinesterase

Introducción: el género Piper perteneciente a la familia Piperaceae es de gran importancia económica debido a sus aplicaciones a nivel alimenticio, industrial y medicinal. Especies de este género son conocidas popularmente como cordoncillos y se caracterizan porque presentan un amplio espectro de actividades biológicas, entre las que se encuentra el efecto neuroprotector, que está asociado al tratamiento de enfermedades neurodegenerativas como el Alzheimer. En Colombia existen muchas especies a las cuales no se les han desarrollado estudios investigativos, tal es el caso de Piper pesaresanum C. DC. Objetivo: caracterizar química y biológicamente el extracto etanólico de hojas de P. pesaresanum mediante un estudio fitoquímico biodirigido y evaluar la actividad inhibitoria sobre acetilcolinesterasa. Métodos: a partir del extracto etanólico de hojas maduras de P. pesarenasum se realizó un estudio químico biodirigido para aislar e identificar las sustancias responsables de la actividad inhibitoria sobre acetilcolinesterasa. La evaluación de la actividad inhibitoria sobre acetilcolinesterasa se realizó por autografía directa empleando el método de Ellman. Adicionalmente se realizó un tamizaje fitoquímico preliminar mediante pruebas de coloración y precipitación llevadas a cabo en vía húmeda y cromatografía en capa delgada. Resultados: el análisis fitoquímico preliminar sugiere la presencia de terpenos, esteroides, fenoles y cumarinas. La evaluación de la actividad inhibitoria sobre acetilcolinesterasa permitió evidenciar zonas claras de inhibición en las fracciones de éter de petróleo (EP) y cloroformo (CHCl3). El estudio fitoquímico biodirigido condujo al aislamiento e identificación por primera vez para la especie del ácido 4-metoxi-3,5-di(3'-metil-2'-butenil) benzoico. Conclusiones: el estudio químico y de actividad biológica llevado a cabo en la especie P. pesaresanum permitió identificar al ácido 4-metoxi-3,5-di(3'-metil-2'-butenil) benzoico como uno de los compuestos responsables de la actividad inhibitoria sobre acetilcolinesteras(AU)

Introduction: The genus Piper, from the Piperaceae family, has great economic importance, due to its uses on a nutritional, industrial and medicinal level. The species in this genus are commonly known as peppers, and are characterized by a broad range of biological activities, including a neuroprotective effect associated with the treatment of neurodegenerative conditions such as Alzheimer's disease. Many of the species growing in Colombia have not been studied. Such is the case with Piper pesaresanum C. DC. Objective: Carry out a chemical and biological characterization of the ethanolic extract of P. pesaresanum leaves by means of a bioguided phytochemical study and evaluate inhibitory activity on acetylcholinesterase. Methods: A bioguided chemical study was conducted of the ethanolic extract of P. pesarenasum mature leaves to isolate and identify the substances responsible for inhibitory activity on acetylcholinesterase. Evaluation of inhibitory activity on acetylcholinesterase was based on direct autography using the Ellman method. Preliminary phytochemical screening was also performed by means of wet color and precipitation tests, and thin-layer chromatography. Results: Preliminary phytochemical analysis suggests the presence of terpenes, steroids, phenols and coumarins. Evaluation of inhibitory activity on acetylcholinesterase revealed clear-cut inhibition areas in petroleum ether (PE) and chloroform (CHCl3) fractions. The bioguided phytochemical study led to isolation and identification of 4-methoxy-3.5-di(3'-methyl-2'-butenyl) benzoic acid for the first time in the study species. Conclusions: Chemical study and evaluation of the biological activity of the species P. pesaresanum led to identification of 4-methoxy-3.5-di(3'-methyl-2'-butenyl) benzoic acid as one of the compounds responsible for inhibitory activity on acetylcholinesterase(AU)

Induction and growth pattern of callus from Piper permucronatum leaves / Indução e padrão de crescimento de calos de folhas de Piper permucronatum

ABSTRACT Piper permucronatum is a perennial shrub, a medicinal plant native to the Amazon Rainforest. Traditionally, the tea of its leaves is used to combat menstrual and intestinal cramps, stomach pain, digestive problems, diarrhea, hemorrhage, and nausea. Its leaf’s essential oil is effective against Aedes aegypti larvae; its flavones and flavanones have a fungicidal effect against Clamidosporium cladosporioides and C. sphaerospermum; its hexanic extract is effective against Leishmania amazonensis. The objective of this study was to provide a protocol for callus induction from P. permucronatum leaves and an identification of the callus growth pattern, focusing on the deceleration phase, when the callus cells must be subcultured into liquid medium in order to produce a cell suspension cultures. Leaf explants were inoculated in a solid MS medium supplemented with factorial combinations of 2,4-D, BA, NAA and GA3. Callus formation was evaluated weekly until the 49th day. Subsequently, new explants were inoculated at the hormonal combination that resulted in the highest callus cell proliferation and, every seven days during a period of 70 days, samples were dried and weighed to determine the callus growth pattern. NAA and GA3 were not effective for callus induction. Combinations of 2,4-D and BA resulted in callus induction and proliferation. The highest percentage of callus induction was observed with the combination of 4.52 µM 2,4-D and 4.44 µM BA. The calluses thereby produced were friable and whitish. The callus growth pattern followed a sigmoid shape. The deceleration phase started on the 56th day of culture.

RESUMO Indução e padrão de crescimento de calos de folhas de Piper permucronatum. Piper permucronatum é um arbusto perene, uma planta medicinal native da Floresta Amazônica. Tradicionalmente, o chá de suas folhas é usado em casos de cólicas menstruais e intestinais, dores de estômago, problemas digestivos, diarreia, hemorragia e náusea. O óleo essencial das folhas é efetivo contra a larva de Aedes aegypti; suas flavonas e flavanonas têm efeito fungicida contra Clamidosporium cladosporioides e C. sphaerospermum; seu extrato hexânico é efetivo contra Leishmania amazonensis. O objetivo deste trabalho foi determinar um protocolo para indução de calos em folhas de P. permucronatum e identificar o padrão de crescimento dos calos, com foco na fase de desaceleração, quando as células de calo devem ser subcultivadas em meio líquido para produzir culturas de células em suspensão. Explantes foliares foram inoculados em meio MS sólido suplementado com combinações fatoriais de 2,4-D, BAP, ANA e GA3. A formação de calos foi avaliada semanalmente até o 49º dia. Posteriormente, novos explantes foram inoculados na combinação hormonal que resultou na maior proliferação de células de calo e, a cada sete dias durante 70 dias, amostras foram secas e pesadas para determinar o padrão de crescimento dos calos. ANA e GA3 não foram efetivas para a indução de calos. Combinações de 2,4-D e BAP resultaram em indução e proliferação de calos. A maior porcentagem de indução de calos foi observada com a combinação de 4,52 µM de 2,4-D e 4,44 µM de BAP. Os calos produzidos eram friáveis e esbranquiçados. O crescimento dos calos seguiu um padrão sigmoide. A fase de desaceleração iniciou no 56º dia de cultivo.

Holopothrips chaconi sp.n. (Thysanoptera: Phlaeothripinae) from leaf-vein galls on Piper species (Piperaceae) in Costa Rica

AbstractMost species of the Neotropical genus Holopothrips are associated with plant galls but very little is known about their biology. Here, we provide observations on the biology of a new species of Holopothrips associated with leaf-vein galls on four species of Piper at a cloud forest site (Zurquí de Moravia) in Costa Rica. This species colonized the galls soon after the gall-inducing cecidomyiid or one of its parasitoids emerged, and several generations of thrips appeared to occupy the empty galls. A total of 175 empty galls from 34 leaves were collected, brought to the laboratory for dissection under the microscope, and the contents quantified. Holopothrips occupied approximately 75 % of the galled leaves and among the latter they occupied about 40 % of the galls. Every combination of adults, nymphs and eggs was found, which implies that adult thrips move in and out of galls, and possibly do not defend their galls from invasion by other members of their species. However, when disturbed, both nymphs and adults raise the tip of their abdomen and emit a distinctive odor, suggesting a defensive reaction against potential predators. Preliminary evidence suggests that they feed on gall tissue lining the inner cavity and spend very little time outside the galls. The thrips is described as a new species, and is one of only four known species of Holopothrips to have the unusual condition of two pairs of epimeral setae on the pronotum. This new species was compared to these other three Holothrips species, and the remarkable variation of the female spermatheca and the male sternal pore plate is illustrated. Further research is needed to confirm that several generations of thrips occupy empty galls, to determine whether adult thrips do indeed move between galls, and to explore in greater detail their possible chemical defense. Rev. Biol. Trop. 63 (4): 1035-1042. Epub 2015 December 01.

ResumenLa mayoría de las especies del género neotropical Holopothrips se asocian a las agallas de plantas pero se sabe muy poco de su biología. Aquí, se provee observaciones sobre la biología de una nueva especie de Holopothrips que se asocia a las agallas de venas foliares en cuatro especies de Piper en un bosque nuboso (Zurquí de Moravia) de Costa Rica. Esta especie coloniza las agallas luego de que el cecidómido inductor o un parasitoide del mismo emerge, y aparentemente varias generaciones de trips ocupan las agallas vacías. Un total de 175 agallas vacías en 34 hojas fueron recolectadas, llevadas al laboratorio para disección bajo el microscopio y se cuantificó su contenido. Holopothrips ocupaba aproximadamente el 75 % de las hojas con agallas y en ellas el 40 % de las agallas. Se encontró toda combinación de adultos, ninfas y huevos, lo cual implica que los trips adultos entran y salen y posiblemente no defienden sus agallas de la invasión de otros miembros de su especie. Sin embargo, adultos y ninfas, levantan la punta del abdomen al ser perturbados, emitiendo un olor distintivo que sugiere una reacción defensiva ante posibles depredadores. La evidencia preliminar indica que se alimentan del tejido interior de la agalla y pasan muy poco tiempo fuera de la misma. Se describe la especie nueva y es una de sólo cuatro especies conocidas de Holopothrips que poseen dos pares de setas epimorales en el pronoto. Al compararla con estas otras especies surge una marcada variación en la espermateca de la hembra y en las placas del poro esternal en el macho, los cuales se ilustran. Se requiere más investigación para confirmar la sugerencia que varias generaciones de trips ocupan las agallas vacias, para determinar si es cierto que los trips adultos se mueven entre agallas, y para explorar en más detalle la supuesta defensa química.

Contribuição ao estudo farmacognóstico da Piper ovatum Vahl / Contribution to the pharmacognostic study of Piper ovatum Vahl

RESUMO: O presente estudo descreve as características morfo-anatômicas dos órgãos vegetativos e do pó da Piper ovatum Vahl de modo que os dados obtidos possam ser utilizados como referência em análises de controle de qualidade de amostras de fármacos, a fim de verificar sua autenticidade. As raízes, caules, pecíolos e folhas foram fixadas, seccionadas à mão livre e coradas, as secções transversais e paradérmicas foram analisadas por microscopia óptica e a superfície do limbo foi observada, também, por microscopia eletrônica de varredura (MEV). Os órgãos vegetativos da P. ovatum apresentam morfologia e anatomia similar às outras espécies de Piper. No entanto, não foram observadas inclusões celulares nas folhas de P. ovatum. Análises por MEV mostraram a presença de tricomas glandulares constituídos de pedúnculo unicelular e porção secretora globóide igualmente unicelular recoberto por cutícula, na epiderme abaxial das folhas. Também foi observada a presença de uma cutícula espessa e que origina crostas no limite entre uma célula e outra, em ambas as superfícies foliares. No mesófilo foi observada a presença de idioblastos oleíferos característica marcante de outras espécies de Piperaceae. Além disso, na microscopia do pó foram observados hipoderme e idioblastos oleíferos em fragmentos do limbo, fragmentos de fibras esclerenquimáticas do caule, além de células esclerosas isoladas ou em grupos no pecíolo. O perfil cromatográfico do extrato hidroetanólico das folhas de P. ovatum foi obtido por cromatografia líquida de alta eficiência (CLAE). Nas análises por CLAE foram identificados como substâncias majoritárias do extrato as amidas piperovatina e piperlonguminina nos tempos de retenção de 10,25 e 10,81 min., respectivamente.

ABSTRACT: The present study describes the morphological and anatomical characteristics of vegetative organs and powder of the Piper ovatum Vahl, in order to use the obtained data as reference in the quality control tests of pharmaceutical samples, investigating their authenticity. The roots, stems, petioles and leaves were fixed, freehand sectioned and stained according to usual microtechniques. The transverse and paradermal sections were analyzed by optical microscopy and the leaf surface was also observed by scanning electron microscopy (SEM). The vegetative organs of the P. ovatum show morphology and anatomy similar to other species of Piper. However, cellular inclusions were not observed in the P. ovatum leaves. The SEM analysis showed the presence of glandular trichomes consisting of a unicellular stalk and globular secretory portion covered by cuticle on the abaxial surface of the leaves. The SEM also had shown one thick cuticle forming crusts in the limit of the epidermal cells, on both leaf surfaces. In the mesophyll, we observed oil idioblasts, which are typical features of other species of Piperaceae. Moreover, in the powder of the P. ovatum we observed hypodermis and oil idioblasts in leaf fragments, fragments of sclerenchyma fibers from the stem and isolated sclereids or in petiole groups. The chromatographic profile of the hydroethanolic extract of the P. ovatum leaves was obtained by high performance liquid chromatography (HPLC). In this analysis, we identified the amides piperovatine and piperlonguminine in the retention times of 10.25 and 10.81 min., respectively, as majority compounds present in the extract.

Efeitos de diferentes tempos de extração no teor e composição química do óleo essencial de folhas de pariparoba [Pothomorphe umbellata (L. ) Miq. ] / Effects of time of extraction in the yield and chemical composition of the essential oil of pariparoba leaves [Pothomorphe umbellata (L. Miq. ]

O presente trabalho visou avaliar a influência de diferentes tempos de extração no teor e composição química do óleo essencial da espécie Pothomorphe umbellata. Folhas de pariparoba foram coletadas em Cajamar - SP, no mês de junho de 2006. O delineamento experimental adotado foi inteiramente casualizado, com seis tempos de extração (90, 120, 150, 180, 210 e 240 minutos) e 4 repetições. A extração do óleo essencial foi realizada por hidrodestilação e a análise química em CG-EM. O tempo de 180 minutos proporcionou maior rendimento do óleo essencial (0,42%), enquanto a composição química não foi influenciada pelos diferentes tempos de destilação. As principais substâncias identificadas foram: D-germacreno (65,5%), alfa-selineno (9,7%), trans-cariofileno (7,5%), e espatulenol (6,58%). Conclui-se que o tempo de extração afeta diretamente o rendimento e a proporção das substâncias químicas, recomendando-se a extração em até 180 minutos. .

The present work aimed to evaluate the influence of different times of extraction in the yield and chemical composition of the essential oil of "pariparoba" (Pothomorphe umbellate). Leaves of "pariparoba" were collected in the municipal district of Cajamar - state of São Paulo, Brazil, in June 2006. The adopted experimental design was completely randomized, with six times of extraction (90, 120, 150, 180, 210, and 240 minutes) and 4 repetitions. The extraction of the essential oil was accomplished by hydrodistillation, and the chemical analysis was performed in GC-MS. The time of extraction of 180 minutes provided greater yield of the essential oil (0.42%), and its chemical composition was not influenced by the different times of hydrodistillation. The main substances showed are D-germacrene (65.5%), alpha-selinene (9.7%), trans-caryophyllene (7.5%) and spathulenol (6.58%). The results of this work showed that the time of extraction affects the yield and the proportion of the chemical substances and it is recommended the extraction in up to 180 minutes.

O extrato bruto hidroalcoólico das folhas de Piper caldense C. DC. na redução do consumo de álcool em ratos / The hydroalcoholic extract of leaves of Piper caldense C. DC. decreases alcohol consumption in rats

A investigação de plantas medicinais como recurso terapêutico para combater o alcoolismo ainda é muito insipiente com poucas espécies investigadas. No presente trabalho avaliamos o efeito do extrato hidroalcoólico das folhas da planta Piper caldense C. DC. com relação ao consumo de álcool e também a sua potencialidade ansiolítica. Foram utilizados ratos Wistar machos, os quais foram divididos em grupos controle (solução fisiológica) e tratado (extrato da planta nas doses de 25, 50 e 150 mg/kg, e diazepam 2 mg/kg). Os animais foram tratados através da via intraperitoneal 1 h antes dos experimentos. Os modelos animais utilizados foram: Labirinto em Cruz Elevado (LCE) e auto-administração de álcool (AA). No LCE o extrato não apresentou efeito ansiolítico do tipo benzodiazepínico. Por outro lado, no comportamento relacionado a auto-administração de solução de álcool, o extrato apresentou efeito significativo reduzindo o consumo de álcool em relação ao grupo controle. Apesar do extrato não ter apresentado efeito ansiolítico, o mesmo parece apresentar potencialidades para combater o abuso e a dependência de álcool.

The investigation of medicinal plants as therapeutic resources in the combat of alcoholism is still very incipient with few species being investigated. The purpose of the present study was to examine the effects of the hydroalcoholic extract of leaves of Piper caldense C. DC. in relation to alcohol consumption and also its anxiolytic potentiality. Male Wistar rats, which had been separated into control group (saline) and treated group (Plant extract doses of 25, 50 and 150 mg/kg and 2 mg/kg of diazepam), were used. Animals were injected intraperitoneally 1 h before the tests. The following models were used: Elevated plus maze (EPM) and alcohol self-administration (ASA). In the EPM, the extract did not show benzodiazepine anxiolytic effect. On the other hand, in the behavior related to alcohol self-administration, the extract showed a significant effect, reducing alcohol consumption compared to the control group. Although the extract has not shown any anxiolytic effect, the results suggest that the plant has potential to combat alcohol abuse and addiction.

Essential oils of leaves of Piper species display larvicidal activity against the dengue vector, Aedes aegypti (Diptera: Culicidae) / Óleo essencial da folha de espécies de Piper exibem atividade larvicida contra o vetor da dengue Aedes aegypti (Diptera: Culicidae)

The mosquito Aedes aegypti is the vector of the dengue virus, an endemic arbovirus from tropical and subtropical regions of the world. The increasing resistance of mosquitoes to commercial insecticides impairs regular control programs; therefore, chemical prospecting originating from the Amazonian flora is promising for potential new insecticides. Several Piper species are, notably, rich in phenylpropanoids and terpenoids, substances with proven insecticidal activity. The composition and the larvicidal activity of three Piper species against A. aegypti were evaluated. Essential oils were extracted by hydrodistillation in a modified Clevenger apparatus and analyzed by GC/MS. The major components found in Piper arboreum were germacrene D (31.83%) and bicyclogermacrene (21.40%); in Piper marginatum: (E)-methyl isoeugenol (27.08%), (E)-anethole (23.98%) and (Z)-methyl isoeugenol (12.01%); and in Piper aduncum: (E)-isocroweacin (29.52%) and apiole (28.62%) and elemicin (7.82%). Essential oils from the Piperaceae species studied resulted in Lethal Concentrations (LC50) of 34-55 ppm, while LC90 was higher than 100 ppm, except for P. marginatum (85 ppm).

O mosquito Aedes aegypti é o vetor do vírus da dengue, um arbovírus endêmico em regiões tropicais e subtropicais do mundo. A crescente resistência dos mosquitos aos inseticidas comerciais prejudica programas regulares de controle, portanto, a prospecção química proveniente da flora amazônica surge como alternativa promissora para novos inseticidas. Várias espécies de Piper são notavelmente ricas em fenilpropanóides e terpenóides, substâncias com atividade inseticida comprovada. A composição e atividade larvicida de três espécies de Piper sobre A. aegypti foi avaliada. Os óleos essenciais foram extraídos por hidrodestilação em Clevenger modificado e analisado por GC/MS. Os principais componentes encontrados em Piper arboreum foram germacreno D (31,83%) e biciclogermacreno (21,40%); Piper marginatum: (E)-metilo de isoeugenol (27,08%), (E)-anetole (23,98%) e (Z)- methyl isoeugenol (12,01%), e Piper aduncum: (E)-Isocroweacin (29,52%), apiol (28,62%) e elemicin (7,82%). Os óleos essenciais a partir das espécies de Piperaceae estudadas resultou em concentrações letais (CL50), 34-55 ppm, enquanto que CL90 foi superior a 100 ppm, com excepção para P. marginatum (85 ppm).

Preferential consumption of larger fruits of Piper arboreum (Piperaceae) by Carollia perspicillata (Phyllostomidae) in the brazilian tropical dry forest / Consumo preferencial de frutos maiores de Piper arboreum (Piperaceae) por Carollia Perspicillata (Phyllostomidae) em uma floresta tropical brasileira

Frugivorous animals may use morphological traits of food items such as size, hardness, shape, color and smell as cues that allow them to assess cost-benefit relationship of foraging activity. Fruit size is an important trait that influences feeding behavior of most frugivores, since there is a functional correlation between fruit size and frugivores' body size. Therefore, size-based preference is fundamental to understand plant-frugivore interactions and seed dispersal. In this sense, we tested the hypothesis of preferential consumption of larger fruits of Piper arboreum by the short-tailed fruit bat Carollia perspicillata in the Brazilian tropical dry forest. Results showed considerable variation in fruit size among and within plants. There was also significant difference in fruit ripening time among plants, which was not related to fruit size. Average size of remaining fruits reduced as they were removed from plants by the bats, which indicates preferential consumption of larger fruits. On the other hand, plant phenology constrained consumption of bigger fruits. Only a small fraction of plant crop was available for consumption each night, regardless of fruit size, which probably coerced bats to feed on smaller fruits as the availability of larger ones decreased. Results suggest that bat preference on fruit size is mediated by plant phenological strategies.

Os frugívoros utilizam as características morfológicas dos frutos tais como tamanho, dureza, forma, cor e cheiro, como pistas para avaliar o custo-benefício da atividade de forrageio. O tamanho dos frutos é uma característica importante que influencia o comportamento alimentar dos frugívoros, já que existe uma correlação funcional entre tamanho do fruto e o do tamanho do frugívoro. Por isso, a seleção do fruto a partir do seu tamanho é um elemento fundamental para a compreensão das interações frugívoro-planta e da dispersão de sementes. Neste sentido, nós testamos a hipótese de consumo preferencial de frutos maiores de Piper arboreum pelo morcego Carollia perspicillata em uma floresta tropical brasileira. Os resultados mostraram ampla variação no tamanho do fruto entre e dentro das plantas parentais. Também houve diferença significativa no tempo de maturação dos frutos entre as plantas, o qual não esteve relacionado com o tamanho do fruto. O tamanho médio dos frutos restantes na planta após a remoção pelos morcegos decresceu, o que indicou o consumo preferencial dos frutos maiores. Por outro lado, a fenologia da planta restringiu o consumo destes frutos. Um número pequeno de frutos maduros esteve disponível para o consumo por noite, independentemente de seu tamanho, o que provavelmente limitou a escolha dos morcegos, obrigando-os a se alimentarem dos frutos menores conforme a disponibilidade dos maiores reduziu-se. Os resultados sugerem que a preferência dos morcegos pelo tamanho dos frutos é mediada pelas estratégias fenológicas das plantas.

Vascular epiphytes in a remnant of seasonal semideciduous forest in zona da mata, State of Minas Gerais, Brazil / Epífitas vasculares em um remanescente de floresta estacional semidecidual na zona da mata, Minas Gerais, Brasil

This study was performed in a remnant of Seasonal Semideciduous Forest at Fazenda Fortaleza de Sant'Anna, in the municipality of Chácara, Minas Gerais, Brazil (22.0129S, 43.8628W), in an area of ca. 1 ha, via monthly expeditions throughout the year 2012. We found 91 species of vascular epiphytes, distributed in 44 genera and 12 families. The richest families were Orchidaceae (35 spp.), Bromeliaceae (18 spp.), Polypodiaceae (10 spp.), Piperaceae (nine spp.) and Araceae (seven spp.). Two species were recorded for the first time in Minas Gerais: Rodriguezia sticta and Stelis oligantha (Orchidaceae). Rhipsalis oblonga (Cactaceae) was recollected in the state after about 100 years without records. Four species are cited in the list of endangered plants in Minas Gerais: Nematanthus crassifolius (Gesneriaceae) and Cattleya bicolor (Orchidaceae) are in the category "Vulnerable" (VU), Nidularium azureum (Bromeliaceae) and Pleurothallis pectinata (Orchidaceae) are "Critically Endangered" (CR). Nidularium azureum is also cited as CR in the Red Book of Brazilian Flora. Most species were classified as characteristic holoepiphytes and anemochory was the most frequent dispersion syndrome.

O estudo foi realizado em um remanescente de Floresta Estacional Semidecidual, na Fazenda Fortaleza de Sant'Anna, município de Chácara, Minas Gerais, Brasil (22,0129S, 43,8628W), em uma área aproximada de 1 ha, através de expedições mensais durante o ano de 2012. Foram encontradas 91 espécies de epífitas vasculares, distribuídas em 44 gêneros e 12 famílias. As famílias mais ricas foram Orchidaceae (35 spp.), Bromeliaceae (18 spp.), Polypodiaceae (10 spp.), Piperaceae (nove spp.) e Araceae (sete spp.). Duas espécies foram registradas pela primeira vez para Minas Gerais: Rodriguezia sticta e Stelis oligantha (Orchidaceae). Rhipsalis oblonga (Cactaceae) foi recoletada no estado depois de cerca de 100 anos sem registros. Quatro espécies estão presentes na lista de plantas ameaçadas de extinção em Minas Gerais: Nematanthus crassifolius (Gesneriaceae) e Cattleya bicolor (Orchidaceae) na categoria "Vulnerável" (VU), Nidularium azureum (Bromeliaceae) e Pleurothallis pectinata (Orchidaceae) como "Criticamente em Perigo" (CR). Nidularium azureum também é citado como CR no Livro Vermelho da Flora do Brasil. A maioria das espécies foi classificada como holoepífita característica e a síndrome de dispersão mais frequente foi a anemocoria.

Avaliação da atividade farmacológica do óleo essencial de Piper rivinoides Kunth / Pharmacological activity of the assessment of the essential oil of Piper rivinoides Kunth

A familia Piperaceae é composta por cinco gêneros e suas espécies são utilizadas para vßrios fins medicinais.Objetivo: investigar o efeito antinociceptivo do óleo essencial de Piper rivinoides Kunth (OEPR). Métodologia:Estudo fitoquímico: Foi utilizado o método MDGC(cromatografia a gßs multi-dimensional) para identificar oscomponentes de OEPR. Modelo de Contorção abdominal: camundongos foram estimulados com ßcido acético0,8 por cento ip. As contorções foram contadas durante 10 minutos. Antagonistas adrenérgicos, imidazolínicos e opióidestambém foram usados neste modelo. Modelo de capsaicina: Capsaicina foi injetada na pata posterior direita (1,6mig/pata). Contou-se o tempo de lambedura por 5 minutos. Teste de formalina: Formalina foi injetada na patadireita (20 mig/pata). A contagem foi realizada em duas fases (1¬ - 5 min e 2¬ - após 15 min a 30 min). Tail Flick:A cauda foi exposta a luz (40 W) e avaliada a retirada da cauda. Teste de placa quente: Observou-se a retirada dapata dos animais tratados. Von frey e edema da pata: Carragenina foi injetada na pata direita (300 mig/pata). 1, 3 e6 horas após o estímulo foram avaliados o número de retirada da pata e o edema. Do macerado do coxim da patados experimentos de Von frey e edema foram dosados proteína e citocinas. Rota rod e habituação em campoaberto: camundongos foram colocados no aparelho de rota rod e verificado o tempo de permanência. No campoaberto foi verificada quantidade de crossing e rearing. Indução de úlcera: Os animais foram tratados com etanola 70 por cento e OEPR e quantificado o número de úlceras. Os animais foram tratados (v.o) com OEPR (0,1, 1, 10, 100e 300 mg / kg) ou veículo 1 hora antes. O tratamento com os antagonistas (ip) foram realizados 20 minutos antesde OEPR, veículo ou clonidina. Morfina e diclofenaco foram os fßrmacos padrões. Todos os experimentos foramrealizados de acordo com o CEUA (033/09), com 5 animais/grupo...

The Piperaceae family consists of five genera and their species are used for various medicinal purposes.Objective: To investigate the analgesic effect of the essential oil of Piper rivinoides Kunth (OEPR).Methodology: Phytochemical studies: We used the MDGC method (multi-dimensional gas chromatography) toidentify OEPR compounds. Writhing Model: Mice were challenged with acetic acid (0.8%, ip). The contortionsare counted for 10 minutes. Adrenergic, imidazoline and opioids antagonists were also used in this model.Capsaicin model: Capsaicin was injected into the hind paw (1.6 µg/paw). It was counted the licking time for 5minutes. Formalin test: Formalin was injected into the right paw (20 µg/paw). Counting was done in two stages(1st one for 5 min and 2nd one 15 min after the first, for 15 min). Tail Flick: The tail was exposed to light (40 W)and the withdrawal of the tail was evaluated. Hot plate test: The paw withdrawal from the hot plate was analyzed.Von frey and paw edema: Carrageenan was injected into the right paw (300 µg/paw). 1, 3 and 6 hours after thestimulation the number of paw withdrawal were assessed and edema. Protein and cytokines from paw cushionmash (Von frey and edema experiments) were measured. Rota rod and Open field: mice were placed on the rodroute set and checked the length of permanence on the instrument. The amount of crossing and rearing wasverified in the open field. Induction of ulcers: Animals were treated with 70% ethanol and OEPR and the numberof ulcers were quantified. The animals were treated (po) with OEPR (0.1, 1, 10, 100 and 300 mg/kg) or vehicle 1hour before. Treatment with antagonists (ip) were conducted 20 min before OEPR, vehicle or clonidine.Morphine and diclofenac were standard drugs. All experiments were performed in accordance with the CEUA(033/09), 5 animals/group...

Propagação vegetativa de estacas de Piper hispidumSw. em diferentes substratos / Vegetative propagation of cuttings of Piper hispidum Sw. in different substrates

RESUMO Piper hispidum é uma espécie pioneira pertencente à família Piperaceae, com importância na medicina popular e na obtenção de óleo essencial. Assim como outras espécies da família, possui poucas informações sobre técnicas de cultivo. O objetivo foi avaliar a propagação via estaquia de Piper hispidum em função do tipo de substrato e estaca em Manaus, Amazonas, Brasil. O experimento foi realizado na Embrapa Amazônia Ocidental. O delineamento utilizado foi inteiramente casualizado em esquema fatorial 3 (estacas) x 5 (substratos), com três repetições de 12 estacas, sendo as estacas (apical, mediana e basal) e os substratos (areia lavada, substrato comercial, solo + esterco de aves, solo + casca de guaraná e fibra de coco). Foram avaliadas: enraizamento (%), número de brotações, comprimento da maior brotação (cm), número de folhas, comprimento da maior raiz (cm), massa seca da raiz (g) e massa seca das brotações (g). Foi realizada análise de variância pelo teste F a 5% de probabilidade e para as médias foi realizado o teste Tukey ao nível de 5% de probabilidade. Recomenda-se utilizar estacas apicais e basais, nesta ordem. Ocorreu interação entre os fatores substrato e tipos de estaca somente para variável número de brotações. Os substratos areia lavada e substrato comercial são indicados para maiores porcentagens de enraizamento com 81,56% e 81,33%, respectivamente. O enraizamento foi superior nas estacas apicais (85,67%) e basais (74,47%). Porém, para esta espécie os substratos solo + esterco de aves e/ou solo + casca de guaraná foram mais indicados quando o objetivo é obter estacas de qualidade.

ABSTRACT Piper hispidum is a pioneer species belonging to the family Piperaceae, with relevance in popular medicine and in obtaining essential oil. As other species of this family, there is little information about cultivation techniques. The aim of this work was to evaluate the propagation through cutting from the Piper hispidum according to the type of substrate and cutting technique in Manaus, Amazonas, Brazil. The trial experiment was conducted at the Embrapa Western Amazon. The design was completely randomized factorial 3 x 5, with three replications of 12 cuttings with types of cuttings (apical, median and basal) and substrates (washed sand, commercial substrate, soil + poultry manure, soil + guarana shell and coconut fiber). After 60 days, the following characteristics were evaluated: rooting (%), number of shoots, length of the largest sprouting (cm), number of leaves, length of the longest root (cm), root dry weight (g) and dry weight of shoots (g). An analysis of variance was performed by the F test at 5% probability and for the averages` comparison the Tukey test was done at 5% level of probability. It is recommended to employ apical and basal cuttings, respectively. There was interaction between the factors and substrate types of cuttings only for the variable number of sprouts. These two substrates, washed sand and commercial substrate, are suggested for higher percentages of rooting with 81,56% and 81,33 %, respectively. The rooting was higher in the apical cuttings (85,67 %) and basal ones (74,47 % ). However, for this species, the substrates soil + poultry manure and/or soil + guaraná shell were most indicated when the goal was to obtain high-quality cuttings.

Circadian variation of essential oil from Piper marginatum Jacq / Variaciones circadiana de aceite esencial de Piper marginatum Jacq

The composition of the essential oil from the leaves of Piper marginatum (Piperaceae) shows circadian variation and a higher yield during high solar incidence. The essential oils samples were investigated by GC-FID and GC-MS, which allowed identification of 29 compounds. Principal component analysis (PCA) and cluster analysis (CA) showed a significant quantitative variability in the chemical composition of the studied samples as well as a correlation between the oil profiles and the collection time. Two main groups were observed, with predominance of phenylpropanoids or sesquiterpenes. The predominant chemical compounds were phenylpropanoids, having as main representative (Z)-asarone and (E)-asarone.

La composición del aceite esencial de hojas de Piper marginatum (Piperaceae) muestra una variación circadiana y un mayor rendimiento durante la mayor incidencia solar. Las muestras de aceites esenciales se investigaron por GC-FID y GC-MS, lo que permitió la identificación de 29 compuestos. El análisis de componentes principales y el análisis de conglomerados mostraron una variación cuantitativa significativa en la composición química de las muestras estudiadas, así como una correlación entre los perfiles del aceite esencial y el tiempo de recolección. Se observaron dos grupos principales, con predominio de fenilpropanoides o sesquiterpenos. Los compuestos predominantes fueron los fenilpropanoides, (Z)-asarona y (E)-asarona.

Avaliação das potenciais atividades tripanocida e antileishmania do extrato de folhas de Piper arboreum (Piperaceae) e de suas frações / Evaluation of the potential trypanocidal and antileishmanial activities in Piper arboreum (Piperaceae) leaf extract and its fractions

A leishmaniose e a tripanossomíase americana fazem parte de um grupo de doenças tropicais endêmicas, especialmente entre as populações pobres. São consideradas doenças negligenciadas por não despertarem interesse da indústria farmacêutica. Atualmente, a quimioterapia é o único tratamento específico disponível para estas doenças, onde os medicamentos utilizados são nifurtimox e benzonidazol. Este trabalho teve como objetivo avaliar a atividade antiparasitária e citotóxica do extrato bruto etanólico e frações de folhas de Piper arboreum, frente a linhagens de Leishmania brasiliensis e de Trypanosoma cruzi. Após a preparação do extrato etanólico bruto e as suas respectivas frações, testes in vitro foram realizados para avaliar atividade antiparasitária frente a T. cruzi, utilizado o clone epimastigota CL-B5 e contra L.brasiliensis foram utilizadas formas promastigotas. O ensaio de citotoxicidade foi realizado com linhagens de fibroblastos NCTC929. Os resultados indicaram que as amostras apresentaram toxidade elevada, com exceção da fração de acetato de etila. Já os resultados da atividade antiparasitária indicaram uma maior atividade da fração hexânica observando a morte de 62% e 51% de células epimastigotas e de 100% e 92% de células promastigotas, respectivamente. Através dos resultados conclui-se que a P. arboreum pode ser considerada uma fonte alternativa de produtos naturais com atividade contra T. cruzi e L. brasiliensis...

Leishmaniasis and South American trypanosomiasis (Chagas disease) are part of a group of related human diseases endemic in tropical regions, especially among the poor. They are referred to as neglected diseases, because of a lack of interest on the part of the pharmaceutical industry. Today, chemotherapy is the only specific treatment against these diseases, the most frequently used drugs being nifurtimox and benznidazole. The aim of this study was to assess the antiparasitic and cytotoxic activity of the crude ethanolic leaf extract of Piper arboreum and its fractions against strains of Leishmania brasiliensis and Trypanosoma cruzi. After preparing the crude ethanolic extract and fractions, in vitro tests were performed to assay antiparasitic activity against T. cruzi, using the epimastigote clone CL-B5, while against L. brasiliensis, promastigote forms were used. The cytotoxicity assay was performed with a fibroblast cell line, NCTC 929. The results indicate that the samples possessed high toxicity, except for the ethyl acetate fraction. The results for antiparasitic activity showed highest activity in the hexane fraction, which was lethal to 62% and 51% of the epimastigotes and 100% and 92% of the promastigotes, at 500 and 100µg/mL, respectively. From these results, it is concluded that P. arboreum can be considered an alternative source of natural products with activity against T. cruzi and L.brasiliensis...