Cyclopeptide Alkaloids from Discaria chacaye (Rhamnaceae) as Result of Symbiosis with Frankia (Actinomycetales).

Cyclopeptide alkaloids with different biological activities are present in plants of the family Rhamnaceae. Plants of this family grow in a symbiotic relationship with aerobic Gram-positive actinomycetes belonging to the genus Frankia. This goal of this research was a study of the comparative profile of alkaloids present in Discaria chacaye and to establish a connection between the presence or absence of Frankia sp. and the alkaloids. In addition, insecticidal activities of the alkaloidal extract were examined. A total of 24 alkaloids were identified, of which 12 have a benzylisoquinoline skeleton, 9 were cyclopeptides, 2 isoquinolines, and 1 aporphine. The presence of cyclopeptide alkaloids is associated with Frankia nodules in the plant root. The alkaloid extracts showed insecticidal activity with mortality dose-dependence and LD50 values between 44 to 71 µg/mL.

Japanese grape (Hovenia dulcis) powder as an antioxidant agent in Bologna sausages.

BACKGROUND: The Japanese grape (Hovenia dulcis Thunberg) has potential as an antioxidant due to its high content of phenolic compounds in the early maturation stages. In this study, the antioxidant sodium erythorbate (CONTROL) was replaced by Japanese grape pseudofruits dehydrated by freeze-drying (FDP) and oven drying (OP), in Bologna sausages. The commercial natural antioxidant licresse (LS), based on licorice, has also been studied. RESULTS: No significant differences (P < 0.05) were observed for the proximate composition and the texture profile of the Bologna sausages. An increase in pH was observed during storage for all formulations. At 28 days, the treatments FDP and OP were darker (lower L* values) and less red (lower a* values), possibly due to the presence of tannins in these samples, which may have bound with the iron present in myoglobin and minimized the oxidation reactions. The use of different antioxidants affected the nitrite contents of the Bologna sausages during storage. Regarding the lipid oxidation, the Japanese grape powder was more effective in delaying the oxidation reactions in Bologna sausages after 14 and 21 days of storage when compared to LS and the CONTROL, respectively. CONCLUSION: The Japanese grape powders were effective in retarding the lipid oxidation of Bologna sausages, possibly due to the presence of bioactive compounds, such as phenolic compounds, tannins, and ascorbic acid, showing the potential of these pseudofruits as antioxidants for use in meat products. © 2022 Society of Chemical Industry.

New cyclopeptide alkaloid of Condalia buxifolia and the absolute stereochemistry of Condaline A.

During the course of a study of Condalia buxifolia (Rhamnaceae), one new cyclopeptide alkaloid condaline B (1), together with six known cyclopeptide alkaloids, condaline A (2), and the scutianines B (3), - D (4) and -E (5), frangulanine (6), and 3,4,28-tris-epi-scutianene N (7), were isolated from the rind bark of Condalia buxifolia. Their structures have been confirmed through spectroscopic analyses such as 1D and 2D NMR experiments. The absolute stereochemistry of condaline A (2), was elucidated by X-ray crystal structure determination of its HI salt. In addition, condaline B (1) was obtained synthetically through a structural transformation of condaline A. Meanwhile, the crude methanol extract, the basic ether fraction, and alkaloids 1-7 were tested against various strains of Gram-positive and Gram-negative bacteria and fungus, showing promising antimicrobial activity.

Alkaloids from the stem barks of Scutia buxifolia Reissek (Rhamnaceae): Structures and antimicrobial evaluation.

A reinvestigation of the chemical constituents of the stem barks of Scutia buxifolia, a member of the Rhamnaceae, resulted, along with the known alkaloids scutianine C and scutianene L, in the isolation of three undescribed diastereoisomeric alkaloids - scutianine N, 27-epi-scutianine N and 3, 4, 7-tri-epi-scutianine N -, one undescribed non macrocyclic alkaloid - scutianine Q - and a neutral compound -scutianene M. Their structures were determined using extensive NMR techniques and HRMS. The absolute configurations of the stereogenic centers of the three diastereoisomeric alkaloids have been assigned by gas chromatography employing modified cyclodextrins as chiral stationary phases. Scutianine Q had its structure and stereochemistry defined by single crystal X-ray crystallographic analysis. All tested compounds showed good to moderate antibacterial activity (MICs between 1.56 and 100 µg mL-1) when evaluated in vitro against a panel of Gram-positive and Gram-negative bacteria. Some stereochemistry-activity relationships have been identified for the antibacterial activity of diastereoisomeric alkaloids against the Gram-negative bacteria Enterobacter aerogenes. The alkaloid 27-epi-scutianine N was as active as the standard antibiotic chloramphenicol (MIC = 1.56 µg mL-1), while scutianine N and 3,4,27-tris-epi-Scutianine N were inactive (>100 µg mL-1).

Phytochemical analysis and evaluation of antioxidant and antimycobacterial activity of Colletia paradoxa from Brazil.

Colletia paradoxa (Spreng.) Esc. (Rhamnaceae, Colletieae) is a medicinal plant, threatened with extinction in Brazil, presenting great morphological variability. Our objective is to investigate the phytochemical components, antioxidant capacity and antimycobacterial activity of different morphotypes of C. paradoxa in different environments. For this, the crude extract of the leaves and branches of the individuals sampled was used. The elimination capacity of the free radicals was determined by the DPPH method, the antimycobacterial activity by the broth microdilution method and the phenolic content by the spectrophotometric method using the Folin-Ciocalteu reagent and by HPLC. The extracts of C. paradoxa and its morphotypes showed significant amounts of phenolic compounds, including quercetin, quercitrin and rutin, besides considerable antioxidant and antimycobacterial activity No connection was detected between the phytochemical composition and different morphotypes of C. paradoxa.

Assessment of Insecticidal Activity of Benzylisoquinoline Alkaloids from Chilean Rhamnaceae Plants against Fruit-Fly Drosophila melanogaster and the Lepidopteran Crop Pest Cydia pomonella.

The Chilean plants Discaria chacaye, Talguenea quinquenervia (Rhamnaceae), Peumus boldus (Monimiaceae), and Cryptocarya alba (Lauraceae) were evaluated against Codling moth: Cydia pomonella L. (Lepidoptera: Tortricidae) and fruit fly Drosophila melanogaster (Diptera: Drosophilidae), which is one of the most widespread and destructive primary pests of Prunus (plums, cherries, peaches, nectarines, apricots, almonds), pear, walnuts, and chestnuts, among other. Four benzylisoquinoline alkaloids (coclaurine, laurolitsine, boldine, and pukateine) were isolated from the above mentioned plant species and evaluated regarding their insecticidal activity against the codling moth and fruit fly. The results showed that these alkaloids possess acute and chronic insecticidal effects. The most relevant effect was observed at 10 µg/mL against D. melanogaster and at 50 µg/mL against C. pomonella, being the alteration of the feeding, deformations, failure in the displacement of the larvae in the feeding medium of D. melanogaster, and mortality visible effects. In addition, the docking results show that these type of alkaloids present a good interaction with octopamine and ecdysone receptor showing a possible action mechanism.

Pollen and microsporangium development in Ziziphus jujuba, Z. mucronata, Paliurus spina-christi and Gouania ulmifolia (Rhamnaceae).

The aim of this paper is to investigate the ultrastructural events that occur during pollen grains development, with emphasis in pollen grain wall and tapetum ontogeny in Ziziphus jujuba, Z. mucronata, Paliurus spina-christi (Paliureae) and Gouania ulmifolia (Gouanieae). Anthers at different developmental stages were processed according to classic techniques for transmission electron microscopy. Differences in the number of endothecium layers and in the number of tapetal cell nuclei were found. Tapetal cells present an anastomosing tubular network and large vesicles with fibrillar content in the cytoplasm. Pollen grain development and ontogeny of pollen grain wall are similar in the four species. The number of endothecium layers, the number of nuclei of the tapetal cells and tapetal cells ultrastructure of the four species support the phylogenetic relationships previously published for the Rhamnaceae family. Tapetal vesicles with fibrillar or polysaccharide content seem to be an exclusive characteristic of the tribes Paliureae and Gouanieae. Some ultrastructural characters of the pollen grain wall development are common to other species of Rhamnaceae, such as the primexine matrix present at the microspore mother cell stage, the aperture entirely built up during the tetrad stage, the thick and fibrillar intine, and the granular infractectum.

Molecular Characterization of Hovenia Dulcis-Associated Virus 1 (HDaV1) and 2 (HDaV2): New Tentative Species within the Order Picornavirales.

In a systematic field survey for plant-infecting viruses, leaf tissues were collected from trees showing virus-like symptoms in Brazil. After viral enrichment, total RNA was extracted and sequenced using the MiSeq platform (Illumina). Two nearly full-length picorna-like genomes of 9534 and 8158 nucleotides were found associated with Hovenia dulcis (Rhamnaceae family). Based upon their genomic information, specific primers were synthetized and used in RT-PCR assays to identify plants hosting the viral sequences. The larger contig was tentatively named as Hovenia dulcis-associated virus 1 (HDaV1), and it exhibited low nucleotide and amino acid identities with Picornavirales species. The smaller contig was related to insect-associated members of the Dicistroviridae family but exhibited a distinct genome organization with three non-overlapping open reading frames (ORFs), and it was tentatively named as Hovenia dulcis-associated virus 2 (HDaV2). Phylogenetic analysis using the amino acid sequence of RNA-dependent RNA polymerase (RdRp) revealed that HDaV1 and HDaV2 clustered in distinct groups, and both viruses were tentatively assigned as new members of the order Picornavirales. HDaV2 was assigned as a novel species in the Dicistroviridae family. The 5' ends of both viruses are incomplete. In addition, a nucleotide composition analysis (NCA) revealed that HDaV1 and HDaV2 have similarities with invertebrate-infecting viruses, suggesting that the primary host(s) of these novel virus species remains to be discovered.

Localization, morphology, anatomy and ultrastructure of osmophores in species of Rhamnaceae.

Although the presence of scent was described for several species of Rhamnaceae, localization, morphology and structure of osmophores were unknown. We studied different species of the tribes Rhamneae (Rhamnoids clade), Pomaderreae, Colletieae, Paliureae (Ziziphoids clade) and the species Alphitonia excelsa (unknown tribe, Ziziphoids clade). We expect to have a better comprehension of these structures and provide information on which morphological and anatomical characters may support the phylogeny of the family. We localized the osmophores in the margins and top of the sepals using neutral red. Histochemical tests were made on transverse hand-cut sections of fresh sepals. Observations were made with stereoscopic and bright field microscopes, scanning and transmission electron microscopes. Papillae were observed in the zones with positive reaction to reagents. Different kinds of hairs are present in the sepal epidermis besides papillae. Epidermal cells present a striate cuticle with canals and cavities. Druses are abundant in most species. The ultrastructure of epidermal and subepidermal cells shows high metabolic activity: there are vesicles, mitochondria, endoplasmic reticulum, dictyosomes, plastids with lipids and starch. The vascularization is well developed and reaches the top of the sepal where the principal area of volatile components production is localized. The location and abundance of papillae are the most important traits that allow us recognize and characterize the osmophores in Rhamnaceae. There are no clear anatomical and morphological features exclusive of one clade or tribe. Therefore, in contrast to other sporophytic structures of this family, osmophores do not seem to have any systematic value.

Características y contenido de extractivos del leño de Discaria chacaye y Ochetophila trinervis (Rhamnaceae) de zonas de ecotono del suroeste de la provincia de Neuquén / Characteristics and wood extract contents of Discaria chacaye and Ochetophila trinervis (Rhamnaceae) from ecotonal areas of southwest Neuquen province

En los ambientes de ecotono entre los bosques y la estepa en Patagonia, destacados por presentar alta biodiversidad y afectados históricamente por diversas actividades humanas, se presentan dos valiosas especies leñosas: Discaria chacaye y Ochetophila trinervis (Rhamnaceae) muy utilizadas para leña y con gran importancia ecológica, social y cultural en la región. Con el objetivo de realizar un aporte al conocimiento del leño de estas especies se aprovechó madera de ejemplares adultos extraídos para leña en cercanías a San Martín de los Andes, suroeste de la provincia de Neuquén, República Argentina. Se elaboraron tablas, cubos, cortes microscópicos y macerados para la determinación de características estéticas, macroscópicas, microscópicas, propiedades físicas (densidad y contenido de humedad de equilibrio higroscópico) y de contenido de extractivos del leño. Los caracteres estéticos y macroscópicos de la madera de las dos especies indican su aptitud para usos en artesanías y trabajos con piezas de poca talla. Las características anatómicas señalan alta seguridad conductiva y especialización del leño de ambas especies a situaciones de estrés hídrico o térmico. La alta densidad de la madera explica en gran parte su preferencia para el uso como combustible en relación a otras especies leñosas de la región. El contenido de extractivos de la madera de ambas especies (14 %) fue levemente mayor al reportado para especies de climas templados. Consideramos que la presente información podrá ser de utilidad para futuros estudios orientados al manejo sustentable o conservación de estas valiosas especies y de los ambientes donde crecen.

In ecotone environments between woodlands and steppe in Patagonia, highlighted by its high biodiversity and historically affected by multiple human activities, two valuable woody species are present: Discaria chacaye and Ochetophila trinervis (Rhamnaceae) widely used for firewood and with great ecological, social and cultural importance in the region. In order to contribute to the knowledge of the bole of these species, wood from adult specimens, extracted for firewood near San Martín de los Andes, southwest of the province of Neuquén, Argentina, was procured. boards, cubes, microscopic cuts and macerations were prepared for determination of aesthetic, macroscopic and microscopic characteristics, physical properties (density and hygroscopic equilibrium moisture content) and wood extracts content. Aesthetic and macroscopic wood characters of both species indicate their aptitude for use in handicrafts and small pieces of manufacture. Wood anatomical characteristics point out high conductive safety and specialization of both species for water and thermal stress situations. The high density in the wood of both species largely explains its preference as a fuel in the region. Wood extractives content of both species was slightly higher than expected for temperate climate species. We believe this information may be useful for future studies aimed at sustainable management and conservation of these valuable species and the environments where they grow.

Methodology proposed for photogrammetric monitoring of the exotic species Hovenia dulcis Thunb. in the Green Belt area surrounding the city of Santa Cruz do Sul, RS, Brazil.

The spread of the exotic species Hovenia dulcis known as Japanese raisin tree, coming from Asia, detected in a protected area of 465.0 ha surrounding the city of Santa Cruz do Sul, RS, Brazil, named "Green Belt." In this context, this research aimed at developing an aerial monitoring system able to identify and quantify the extension in the Green Belt area that this species invaded, as well as describing the impacts caused on the local forest community. We collected data from an ultralight Flyer GT aircraft equipped with a vertical camera. The images were taking in June and July 2015, when H. dulcis trees have shed their leaves, displaying a gray color, and September and October 2015, when the leaves are young and with a light green color. Based on the methodology proposed trough aerial monitoring by photogrammetry, the results indicated that the total invaded area by H. dulcis in the Green Belt of Santa Cruz do Sul County, RS, was 131.8 ha, corresponding to 20.9% of the total area. To validate this method, a quantitative comparison between the result from a terrestrial fieldwork carried out and the proposed method showed no significant differences in the estimated area occupied by H. dulcis. We concluded that these results validate the proposed aerial assessment method.

Vagrant birds as a dispersal vector in transoceanic range expansion of vascular plants.

Birds are thought to be important vectors underlying the disjunct distribution patterns of some terrestrial biota. Here, we investigate the role of birds in the colonisation by Ochetophila trinervis (Rhamnaceae), a vascular plant from the southern Andes, of sub-Antarctic Marion Island. The location of O. trinervis on the island far from human activities, in combination with a reconstruction of island visitors' travel history, precludes an anthropogenic introduction. Notably, three bird species occurring in the southern Andes inland have been observed as vagrants on Marion Island, with the barn swallow Hirundo rustica as the most common one. This vagrant displays long-distance migratory behaviour, eats seeds when insects are in short supply, and has started breeding in South America since the 1980s. Since naturalised O. trinervis has never been found outside the southern Andes and its diaspores are incapable of surviving in seawater or dispersing by wind, a natural avian dispersal event from the Andes to Marion Island, a distance of >7500 km, remains the only probable explanation. Although one self-incompatible shrub seems doomed to remain solitary, its mere establishment on a Southern Ocean island demonstrates the potential of vagrancy as a driver of extreme long-distance dispersal of terrestrial biota.

Antibacterial activity of Discaria americana Gillies ex Hook (Rhamnaceae).

ETHNOPHARMACOLOGY RELEVANCE: Discaria americana Gillies ex Hook (sin. Discaria febrifuga and Discaria longispina) (Rhamnaceae) is a plant native from Rio Grande do Sul (Southern Brazil), Uruguay and Argentine, and has been used in Brazilian traditional medicine as antipyretic agent, and for stomach disorders. In Rio Grande do Sul, Uruguay and Argentine, the roots, in decoction, are used as tonic and febrifuge. Although it is a plant widely used by the population, there are no studies proving this popular use. MATERIAL AND METHODS: The crude neutral methanol extract, and pure isolated alkaloids, were investigated in vitro for antimicrobial activities against four Gram-positive bacteria: Staphylococcus aureus, Bacillus subtillis, Bacillus cereus, Enterococcus faecium; and five Gram-negative bacteria: Escherichia coli, Enterobacter cloacae, Enterobacter aerogenes, Salmonella enterica serovar Typhimurium and Pseudomonas aeruginosa. RESULTS: The crude neutral methanol (CME) extract of the root bark of Discaria americana showed antibacterial activity, ranging from 62.5 to 250 µg mL-1 (MIC), against the tested bacteria. From the fractions obtained from the crude extract, the basic ethereal fraction (BEF) showed to be more effective, with MICs between 31.5 and 125 µg mL-1 against the tested bacteria. The bioassay-guided fractionation of the ethyl ether basic fraction yielded eight cyclopeptide alkaloids: frangufoline (1), frangulanine (2), adouetine Y' (3), discarine A (4) discarine B (5), discarine C (6), discarene C (7) and discarine D (8). When evaluated against the Gram-positive bacteria Enterococcus faecium, discarine B (5) proved to be the most active alkaloid with a MIC/MLC = 0.77/1.55 µg mL-1, near the most active antibacterial agent levofloxacin (MIC/MLC = 0.77/0.77 µg mL-1). Moreover, discarine C (6) was the more active alkaloid against Salmonella enterica serovar Typhimurium, with a MIC/MLC = 3.1/6.2 µg mL-1, the same observed for the antibacterial agent azithromycin. Kinetic measurements of the bacteriolytic activities of discarine B (5) against Enterococcus faecium (Gram-positive), and of discarine C (6) against Salmonella enterica serovar Typhimurium (Gram-negative) were determined by optical density based on real time assay, suggesting that both mode of action are partially bacteriolytic. CONCLUSION: In conclusion, five 14-membered cyclopeptide alkaloids isolated from Discaria americana Gillies ex Hook (Rhamnaceae) showed promising antibacterial activity, making this metabolites a class of scientific interest. The good activity presented by the extract and the alkaloids against the Gram-positive bacteria Enterococcus faecium and against the Gram-negative bacteria Salmonella enterica serovar Typhimurium, Enterobacter. aerogenes and Escherichia coli, corroborate with the popular use of this plant for stomach disorders and as antifebrile.

Root bark of Discaria americana attenuates pain: A pharmacological evidence of interaction with opioidergic system and TRP/ASIC channels.

ETHNOPHARMACOLOGICAL RELEVANCE: Discaria americana (Rhamnaceae) root bark infusion have been used in traditional medicine as antipyretic, tonic, ameliorative of stomach and skin diseases and diabetes. This study was designed to investigate whether the methanolic extract of the root bark of Discaria americana (MEDa) exhibits antinociceptive effects in mice. Furthermore, it was investigated the involvement of the opioidergic system in MEDa mechanism of action as well the interactions with TRP/ASIC channels in its effect. MATERIALS AND METHODS: The antinociceptive effect of intra-gastric gavage (i.g.) of MEDa (0.3-300 mg/kg) was evaluated in mice subjected to acute chemical (acetic-acid, formalin, glutamate, capsaicin, cinnamaldehyde, and acidified saline) or thermal (hot plate) tests of pain. The involvement of opioid system was evaluated in the formalin test. A nonspecific effect of MEDa was observed by measuring locomotor activity and exploratory behavior in open field test. RESULTS: MEDa significantly reduced the number of writhing induced by acetic acid and inhibited the nociception in the two phases of formalin. These effects were inhibited by pretreatment with naloxone. The nociception induced by hot plate and intraplantar injection of glutamate, capsaicin, cinnamaldehyde and acidified saline were significantly inhibited by MEDa. Only the dose of 300 mg/kg altered the locomotor activity. CONCLUSIONS: Our results demonstrated, for the first time, that the methanolic extract of the root bark of Discaria americana presents antinociceptive effect in chemical and thermal stimuli and its analgesic properties can be due activation of the opioidergic system. These results support the use of Discaria americana in traditional medicine and demonstrate that this plant presents a therapeutic potential for the development of phytomedicines with antinociceptive profile.

Structure of the style and pollen tube pathway in the Ziziphoid and Rhamnoid clades of Rhamnaceae.

The ultrastructure of the style and pollen tube pathway before, during and after anthesis were studied in 13 species belonging to the tribes Pomaderreae, Paliureae, Colletieae and Gouanieae (Ziziphoid clade) and Rhamneae (Rhamnoid clade) using light microscopy and transmission electron microscopy. The aim of this study is to provide new morphological characters useful for phylogenetic analysis at suprageneric level in Rhamnaceae. The patterns of pollen tube growth and the ultrastructural changes undergone by cells of the style were also described. Species of Rhamneae (Scutia buxifolia and Condalia buxifolia) have a solid style, with the transmitting tissue forming three independent strands (S. buxifolia) or a central, single horseshoe-shaped strand as seen in transversal section (C. buxifolia) which could derive from the fusion of formerly independent strands. In contrast, Pomaderreae, Gouanieae and Paliureae showed semi-solid styles, while in Colletieae, as previously reported, the style is hollow with two or three stylar canals. The style anatomy and the ultrastructure of the pollen tube pathway show that there is a tendency towards a solid style with a single strand of transmitting tissue within the family. The three-canalled hollow style could be the plesiomorphic state of the character "type of style" in the family, the semi-solid style the synapomorphic state and the solid style with three strands of transmitting tissue the apomorphic state, with the solid style with a single strand of transmitting tissue as the most derived state. Therefore, Colletieae would be the most basal tribe of the Ziziphoid clade.

Countercurrent chromatography separation of saponins by skeleton type from Ampelozizyphus amazonicus for off-line ultra-high-performance liquid chromatography/high resolution accurate mass spectrometry analysis and characterisation.

Ampelozizyphus amazonicus Ducke (Rhamnaceae), a medicinal plant used to prevent malaria, is a climbing shrub, native to the Amazonian region, with jujubogenin glycoside saponins as main compounds. The crude extract of this plant is too complex for any kind of structural identification, and HPLC separation was not sufficient to resolve this issue. Therefore, the aim of this work was to obtain saponin enriched fractions from the bark ethanol extract by countercurrent chromatography (CCC) for further isolation and identification/characterisation of the major saponins by HPLC and MS. The butanol extract was fractionated by CCC with hexane - ethyl acetate - butanol - ethanol - water (1:6:1:1:6; v/v) solvent system yielding 4 group fractions. The collected fractions were analysed by UHPLC-HRMS (ultra-high-performance liquid chromatography/high resolution accurate mass spectrometry) and MSn. Group 1 presented mainly oleane type saponins, and group 3 showed mainly jujubogenin glycosides, keto-dammarane type triterpene saponins and saponins with C31 skeleton. Thus, CCC separated saponins from the butanol-rich extract by skeleton type. A further purification of group 3 by CCC (ethyl acetate - ethanol - water (1:0.2:1; v/v)) and HPLC-RI was performed in order to obtain these unusual aglycones in pure form.

Antinociceptive and anti-inflammatory effect of the Scutia buxifolia Reissek stem barks extract.

BACKGROUND: Scutia buxifolia (Rhamnaceae) has been extensively studied for its phenolics groups, which are able to capture free radicals; being therefore, considered promising as an antioxidant in preventing diseases resulting from oxidative stress. HYPOTHESIS: Scutia buxifolia extract (SBE) presents antinociceptive and anti-inflammatory effect in mice. STUDY DESIGN: SBE (400-800mg/kg) was tested in different pain models to investigate its antinociceptive and anti-inflammatory action. METHODS: It was carried out the abdominal writhing test, capsaicin test, thermal hyperalgesia and incisional pain. The inflamed tissue by carrageenan was used for the analysis of interleukins (IL), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), c-reactive protein (CRP), nitrite and nitrate (NOx) determination and myeloperoxidase (MPO) activity. Furthermore, we evaluate the possible action mechanism of SBE using naloxone in capsaicin test. RESULTS: SBE prevented the nociception caused by acetic acid, formalin and capsaicin test. However, neither the SBE prevented the thermal hyperalgesia in hot-plate test, nor the naloxone reversed the SBE antinociceptive effect in capsaicin test. Furthermore, the administration of SBE prevented significantly the increase of MPO activity, the NOx content, and the levels of IL-1, IL-6, TNF-α, INF-γ and CRP and was able to increase the IL-10 levels after the inflammation induced by carrageenan in mice. In addition, SBE prevented mechanical hyperalgesia in a postoperative pain model. CONCLUSION: The SBE presents great antinociceptive and anti-inflammatory activity in mice but this effect not seem to have its action mechanism like opioids. It is possible that its antinociceptive effects are associated with levels decrease of inflammatory mediators.

Actividad insecticida y reguladora del crecimiento de extractos de blachnum chillones (blechnaceae) y condolía microphylla cav. (rhamnaceae), sobre larvas de gallería melonada (L.) (lepidoptera: pyralidae) / Insecticidal activity and growth regulatory of blechnum chilense (blechnaceae) and condalia microphylla cav. (rhamnaceae) extracts, on larvae of galleria mellonella (linnaeus) (lepidoptera: pyralidae)

Galleria mellonella (L.) es una de las plagas más importantes para la apicultura, debido a que en su estado larval se alimentan de la cera, polen y miel almacenados en los panales de Apis mellifera (L.). El objetivo del presente estudio fue determinar la actividad insecticida y reguladora del crecimiento de extractos obtenidos a partir de hojas y tallos de Blechnum chilense (Kaulf.) Mett y Condalia microphylla Cav. sobre larvas (L2) de G. mellonella. Los extractos fueron obtenidos con solventes orgánicos de diferente polaridad. Los resultados obtenidos indican que, el extracto de B. chilense obtenido con acetato de etilo y el extracto de C. microphylla obtenido con acetona, tienen efecto insecticida efectivo sobre larvas de G. mellonella y a aplicado en dosis sub-letales afectan negativamente la ganancia de peso larvario. A su vez, el extracto obtenido de B. chilense presentó actividad reguladora del desarrollo larvario de G. mellonella, induciendo el estado de pupa en forma prematura.

Galleria mellonella (L.) is the most detrimental pest to beekeeping, due the larvae feeds on hive of Apis mellifera (L.) consuming the wax, pollen and honey. The aim of this study was to determine the insecticidal activity and growth regulatory activity of extracts obtained from leaves and stems of Blechnum chilense (Kaulf.) Mett and Condalia microphylla Cav. for larvae of G. mellonella. The extracts were obtained with organic solvents of different polarity. The results show that the extract of B. chilense obtained with ethyl acetate, and the extract of C. microphylla obtained with acetone have effective insecticidal activity on larvae of G. mellonella, when applied at sub-lethal doses affect adversely the larval weight gain. In turn, the extract obtained from B. chilense showed regulatory activity on larval development of G. mellonella, inducing pupal stage prematurely.

Inhibitory effect of Scutia buxifolia extracts, fractions, and ursolic acid on Na(+), K(+)-ATPase activity in vitro in membranes purified from rat hearts.

ETHNOPHARMACOLOGICAL RELEVANCE: Scutia buxifolia is a tree native to South America and is used as a cardiotonic agent; however, this property has not been associated with a clear mechanism or a specific compound. AIM OF THE STUDY: Given the importance of Na(+),K(+)-ATPase as a drug target in the treatment of heart failure, this study aimed to investigate the possible inhibitory effect of S. buxifolia crude extract and fractions (dichloromethane, ethyl acetate, and butanolic fractions), and identified compounds with effects on the activity of Na(+),K(+)-ATPase in vitro. MATERIALS AND METHODS: First, we characterized the crude extract and fractions by high-performance liquid chromatography, and then monitored their effects on the activity of Na(+),K(+)-ATPase obtained from heart muscle and brain membranes of adult male Wistar rats. RESULTS: We identified gallic acid, chlorogenic acid, caffeic acid, rutin, quercitrin, quercetin, and ursolic acid in S. buxifolia stem bark and leaves; quercitrin and ursolic acid were the main compounds in the ethyl acetate and dichloromethane fractions from leaves and stem bark. The crude extract (3 and 30mg/ml), and the ethyl acetate and dichloromethane fractions (0.1 and 1mg/ml) of both the stem bark and leaves inhibited Na(+),K(+)-ATPase activity in heart and brain samples. We found that, of the identified compounds, only ursolic acid (0.1mg/ml) was able to diminish Na(+), K(+)-ATPase activity in heart and brain samples. CONCLUSIONS: These data indicated that the cardiotonic effects of S. buxifolia may be due to the inhibition of Na(+),K(+)-ATPase activity in heart muscle, supporting the popular use of this plant as a treatment for heart failure.

Pollen and microsporangium development in Hovenia dulcis (Rhamnaceae): a different type of tapetal cell ultrastructure.

Despite that there is some literature on pollen morphology of Rhamnaceae, studies addressing general aspects of the microsporogenesis, microgametogenesis, and anther development are rare. The aim of this paper is to describe the ultrastructure of pollen grain ontogeny with special attention to tapetum cytology in Hovenia dulcis. Anthers at different stages of development were processed for transmission and scanning electron microscopy, bright-field microscopy, and fluorescence microscopy. Different histochemical reactions were carried out. The ultrastructural changes observed during the development of the tapetal cells and pollen grains are described. Large vesicles containing carbohydrates occur in the tapetal cell cytoplasm during the early stages of pollen development. Its origin and composition are described and discussed. This is the first report on the ontogeny and ultrastructure of the pollen grain and related sporophytic structures of H. dulcis.