Micropropagação, propagação por sementes e banco de germoplasma de Mandevilla velutina (Mart.) Woodson

Mandevilla velutina (Mart.) Woodson (Apocynaceae) é uma planta medicinal de espécie nativa de regiões do Cerrado brasileiro com propriedades anti-ofídicas. Devido a alta exploração e devastação deste bioma, M. velutina está em perigo de extinção. Como uma iniciativa para a conservação desta espécie ameaçada e economicamente importante, um protocolo de micropropagação foi desenvolvido e genótipos foram colocados no Banco de Germoplasma "Cerrado In vitro". Para a propagação in vitro de M. velutina, segmentos nodais foram inoculados no meio Murashige and Skoog (MS) suplementado com diferentes concentrações de BA, zeatina, 2ip, DTT e TDZ. A melhor resposta de multiplicação foi obtida em meio suplementado com 0.44 µM BA na razão 1: 6.7. Plântulas cultivadas em MS/2 suplementado com 26.85 µM NAA enraizaram com sucesso (50.5%). Quando plântulas enraizadas e não enraizadas foram aclimatizadas em condições de solo, grande perda foi observada entre plântulas não enraizadas, enquanto as enraizadas tiveram 100% de sobrevivência. Foi possível a manutenção de 43% de M. velutina em condições saudáveis por seis meses, sem subculturas usando meio MS suplementado com 2% de sacarose, 13.8 mM de espermidina, 2% de sorbitol e 2% de dextrose.Mandevilla velutina (Mart.) Woodson (Apocynaceae) is a medicinal plant species with antivenom properties, native from Brazilian Savanna regions (Cerrado), which due to overexploitation and habitat deforestation is in danger of extinction. As an initiative for conserving this endangered but economically important plant species, a micropropagation protocol was developed and genotypes were stored in the Germplasm Bank "Cerrado In vitro". For the in vitro propagation of M. velutina, nodal segments were inoculated on Murashige and Skoog (MS) medium supplemented with different concentrations of BA, Zeatin, 2ip, DTT and TDZ. Best multiplication ratio was achieved when to the medium 0.44 µM BA, ranging 1: 6.7, were added. Plantlets cultured on MS/2 medium supplemented with 26.85 µM NAA rooted successfully (50.5%). Although rooted and un-rooted plantlets acclimatized to soil conditions, great losses were observed within un-rooted plantlets, while the rooted presented 100 % survival. It was possible to maintain 43% of the M. velutina germplasm under healthy conditions for six months, with no subcultures, using the MS medium supplemented with 2% sucrose, 13.8 mM spermidine, 2% sorbitol and 2% dextrose

Germinação de sementes in vitro e desenvolvimento de plantas ex vitro de algodãozinho-do-campo

Este estudo teve por objetivo avaliar a assepsia, quebra de dormência e germinação das sementes de Cochlospermum regium in vitro e o desenvolvimento ex vitro de plantas dessa espécie. Na assepsia das sementes, foram utilizados benlate, hipoclorito de cálcio e ácido sulfúrico. A superação da dormência foi realizada com sementes imersas em ácido sulfúrico por diferentes períodos. Para verificar a influência do tamanho das sementes na germinação, estas foram classificadas em grandes (A=34,08mm²), médias (A=27,74mm²) e pequenas (A=17,99mm²). No desenvolvimento de plantas ex vitro, foram utilizados substratos areia ou solo de Cerrado+esterco. Eficiente assepsia das sementes foi obtida com a utilização do ácido sulfúrico. A imersão por 146 e 144 minutos, para quebra de dormência, proporcionou maior porcentagem de germinação (63%) e IVG (0,78) respectivamente - sem nenhuma contaminação. As sementes maiores apresentam maior IVG (0,61) em relação a sementes menores. Embora o desenvolvimento das plantas em solo de Cerrado+esterco ou areia tenha sido semelhante, no primeiro substrato, as mudas mostraram-se mais vigorosas.This study was designed to conduct experiments with germination and dormancy breaking in in vitro seeds of Cochlospermum regium in vitro and evaluate the development of plants ex vitro for this species. For the sterilization of seeds introduced in vitro, were used benlate, calcium hypochlorite and sulfuric acid. The break dormancy was realized with seeds immersed in sulfuric acid for different periods. To check the influence of seed size on germination, they were classified as large (A=34.08mm²), medium (A=27.74mm²) and small (A=17.99mm²). For the development of ex vitro plants were used sand substrate or Cerrado soil + manure. Efficient sterilization of seeds was achieved with the use of sulfuric acid. The immersion for 146 and 144 minutes, to break dormancy, showed higher germination rate and germination speed of 63% and 0,78 respectively - with no contamination. Larger seeds germinated faster (0.61) than smaller ones. Though ex vitro development of plants cultured in both substrates was similar, plantlets cultured in soil + manure were more vigorous

Morphologic and morphometric analysis of the root canal apical third cleaning after biomechanical preparation using 3.3% Ricinus communis detergent and 1% NaOCl as irrigating solutions

This study evaluated, by morphologic and morphometric analyses, the cleaning of apical third of root canals instrumented with nickel-titanium rotary files using different irrigating solutions. Twenty-seven single-rooted mandibular premolars were assigned to three groups (n=9), according to the irrigating solution used: Group I, distilled and deionized water; Group II, 1% NaOCl; and Group III, 3.3% Ricinus communis detergent. Biomechanical preparation was performed with Protaper Plusâ nickel-titanium files as follows: S1, SX and S2 at the cervical and middle thirds, and 25/02, 25/04, 25/06, 30/02, 30/04 and 30/06 to complete the instrumentation, operating at 1 mm from the root apex. Irrigation was done at each file change with 2 mL of irrigating solution, totalizing 20 mL for each tooth. After biomechanical preparation, the apical thirds were serially sectioned and histologically processed. The cross-sections were examined by an optical microscope (X40) connected to a computer. The images were captured and analyzed using a computer software and submitted to morphometric analysis with aid of a grid. The percentage of debris remaining at the apical third was calculated. Data were submitted to statistical analysis by ANOVA and Tukey test. The results showed no statistically significant difference (p>;0.01) between the groups irrigated with 1% NaOCl and 3.3% Ricinus communis detergent, which presented the lowest percentage of debris at the apical third, 8.49% and 10.11%, respectively. The group irrigated with distilled and deionized water had the highest percentage of debris (15.58%) and was statistically different from the other groups (pEstudou-se, por meio de análise morfológica e morfométrica, a capacidade de limpeza promovida pela instrumentação rotatória com limas Ni-Ti e irrigação com diferentes soluções. Vinte e sete pré-molares inferiores foram distribuídos em três grupos (n=9), de acordo com a solução irrigante utilizada: Grupo I, água destilada e deionizada; Grupo II, NaOCl a 1% e Grupo III, detergente derivado do óleo de mamona (Ricinus communis) a 3,3%. O preparo biomecânico foi realizado com sistema rotatório de Ni-Ti Protaper Plus, obedecendo a seguinte ordem de uso dos instrumentos: S1, SX e S2 para o terço cervical e médio, seguida do 25/02, 25/04, 25/06, 30/02, 30/04 e 35/02, completando a instrumentação todos operando 1 mm aquém do ápice. A irrigação foi realizada a cada troca de instrumento com 2 mL de solução irrigante, totalizando um volume de 20 mL para cada dente. Após o preparo biomecânico, os terços apicais dos dentes foram submetidos ao processamento histológico. Os espécimes foram analisados em microscópio óptico (40X) conectado a um computador. As imagens foram capturadas e analisadas utilizando-se softwares específicos e submetidas à análise morfométrica por meio de uma grade de integração. A porcentagem de debris presente no terço apical dos canais foi calculada. Os dados foram submetidos à análise estatística por meio de ANOVA e teste de Tukey. Os resultados do presente estudo evidenciaram que não houve diferença estatisticamente significante (p>;0,01) entre os grupos irrigados com NaOCl a 1% e detergente de mamona a 3,3%, que apresentaram menor porcentagem de debris no terço apical, 8,49% e 10,11% respectivamente. O grupo irrigado com água destilada e deionizada apresentou a maior porcentagem de debris (15,58%), sendo estatisticamente diferente (

Chemical constituents from Tabernaemontana catharinensis root bark: a brief NMR review of indole alkaloids and in vitro cytotoxicity

This work describes the isolation and structural determination of pharmacological compounds present in the bark of roots of Tabernaemontana catharinensis (Apocynaceae). Among the 27 substances detected 12 were identified as terpenoid-indole alkaloids, 2 steroids and 13 pentacyclic triterpenes. Structures were outlined based on HMQC, COSY, DEPT, 13C, and ¹H NMR data and MS. Spectral data of indole alkaloids were reviewed. An in vitro screening of the extracts and isolated compounds was carried out. Compounds ibogamine (5), 3-oxo-coronaridine (9) and 12-methoxy-4-methylvoachalotine (MMV) demonstrated effective cytotoxicity towards SKBR-3 breast adenocarcinoma and C-8161 human melanoma tumor cell lines.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNAER

Genetic variability in natural populations of Zeyheria montana mart. from the Brazilian Cerrado

Zeyheria montana, an endemic species of the Bignoniaceae family from the Brazilian Cerrado's known for its anti-cancer properties, is widely used as imuno stimulant in the popular medicine and its therapeutic activity must be validated by scientific data. The objective of this work was to evaluate the genetic variability of eight plant populations collected within the state of São Paulo, Brazil, via Random Amplification of Polymorphic DNA (RAPD) used as molecular markers. After an optimized protocol for the amplification reaction, nine selected primers generated 105 reproducible bands, indicating up to 60% polymorphism. Analysis of molecular variance (AMOVA) revealed higher genetic variation within populations (84.03%) than among populations (15.97%). The variation values estimated by phiST (0.160) indicated moderate to high inter population structuration. Levels of similarity inter plants with genetic and geographical distances, estimated by the unweighted pair-group method analysis (UPGMA) clustering and non-metric multidimensional scaling (NMDS) ordination methods and by the Mantel test (-0.2345 p = 0.118) denoted that the structure found follows the island model, which assumes that a single population of infinite size may have initiated the existing populations of Zeyheria montana, with no spatial position correlation. Based on the obtained data, a germplasm bank from individuals representing the species variability was established. Furthermore the information here reported can be of importance to develop strategies for the conservation of Z. montana.Zeyheria montana, planta arbustiva da família Bignoniaceae, é uma espécie endêmica do Cerrado e possui atividade anti-câncer, sendo utilizada como estimulante na medicina popular. O objetivo deste estudo foi avaliar a variabilidade genética de oito populações localizadas no estado de São Paulo, utilizando marcadores moleculares de Polimorfismo de DNA Amplificado ao Acaso (RAPD). Após a otimização da reação de amplificação, nove iniciadores selecionados geraram 105 fragmentos RAPD reprodutíveis, sendo que a maioria (60,0%) foi polimórfica. A análise molecular de variância (AMOVA) mostrou que a variabilidade dentro de populações (84,03%) foi maior que entre populações (15,97%). As estimativas de variação fiST (0,1597) indicam estruturação populacional moderadamente alta. O agrupamento por meio de UPGMA, a ordenação pelo NMDS e o teste de Mantel entre as matrizes de distâncias genéticas e geográficas demonstraram que a estruturação encontrada segue um modelo de "ilhas", onde uma única população de tamanho infinito pode ter dado origem às populações atuais de Zeyheria, sem relação com sua posição espacial. Com base nos resultados obtidos foi estruturado um banco de germoplasma de indivíduos, representando a variabilidade da espécie. Adicionalmente, as informações deste estudo são importantes para dar suporte a estratégias de conservação de Z. montana

Preservation of phenolic compounds on dried leaf infusion of Bauhinia forficata Link

Bauhinia forficata Link aqueous extract is usually recommended as a phytomedicine to reduce blood glucose levels and its biological activity has been linked to the presence of phenolic compounds from B. forficata preparations. Several drying processes are used in the production of dry herbal extracts, which may influence the chemical composition and efficacy of final herbal medicines. Due to significant chemical changes, defining appropriate drying processes is essential for phytopharmaceutical drug development. In view of this, we analyzed dried B. forficata leaf infusion (BFLI) extracts by HPLC-UV-MSn, followed by molecular networking analysis to evaluate the chemical profiles from dried extracts yielded by freeze-and spray-drying processes. The main metabolites detected included 11 ferulic/isoferulic acid derivatives and 13 glycosylated flavonoids. The qualitative chemical profiles were alike for both drying processes, whereas the relative abundance of some flavonoids was higher using spray-drying. Taken together, our results showed that freeze-and spray-drying preserved the phenolic profile of BFLI and suggested that spray-drying may be the most suitable to obtain its dried products. Along with studying the chemical profiles of dried herbal extracts, evaluating the influence of drying processes on the quality and chemical profiles of final products is pivotal and may benefit future research

Arcuate AgRP, but not POMC neurons, modulate paraventricular CRF synthesis and release in response to fasting

Background: The activation of the hypothalamic–pituitary–adrenal (HPA) axis is essential for metabolic adaptation in response to fasting. However, the neurocircuitry connecting changes in the peripheral energy stores to the activity of hypothalamic paraventricular corticotrophin-releasing factor (CRFPVN) neurons, the master controller of the HPA axis activity, is not completely understood. Our main goal was to determine if hypothalamic arcuate nucleus (ARC) POMC and AgRP neurons can communicate fasting-induced changes in peripheral energy stores, associated to a fall in plasma leptin levels, to CRFPVN neurons to modulate the HPA axis activity in mice. Results: We observed increased plasma corticosterone levels associate with increased CRFPVN mRNA expression and increased CRFPVN neuronal activity in 36 h fasted mice. These responses were associated with a fall in plasma leptin levels and changes in the mRNA expression of Agrp and Pomc in the ARC. Fasting-induced decrease in plasma leptin partially modulated these responses through a change in the activity of ARC neurons. The chemogenetic activation of POMCARC by DREADDs did not affect fasting-induced activation of the HPA axis. DREADDs inhibition of AgRPARC neurons reduced the content of CRFPVN and increased its accumulation in the median eminence but had no effect on corticosterone secretion induced by fasting. Conclusion: Our data indicate that AgRPARC neurons are part of the neurocircuitry involved in the coupling of PVNCRF activity to changes in peripheral energy stores induced by prolonged fasting.Fil: Alves Fernandes, Alan Carlos. University Of Ribeirao Preto; BrasilFil: Pereira de Oliveira, Franciane. Universidade Federal de Sao Paulo.; BrasilFil: Fernandez, Gimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: da Guia Vieira, Luane. University Of Ribeirao Preto; BrasilFil: Gugelmin Rosa, Cristiane. University Of Ribeirao Preto; BrasilFil: do Nascimento, Taís. University Of Ribeirao Preto; BrasilFil: de Castro França, Suzelei. University Of Ribeirao Preto; BrasilFil: Donato Jr, Jose. Universidade de Sao Paulo. Departamento de Fisiología; BrasilFil: Vella, Kristen R.. Weill Cornell Medical College; Estados UnidosFil: Antunes Rodrigues, Jose. University Of Ribeirao Preto; BrasilFil: Mecawi , André. Universidade Federal de Sao Paulo.; BrasilFil: Perello, Mario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Leico Kagohara Elias, Lucila. University Of Ribeirao Preto; BrasilFil: Rorato, Rodrigo. Universidade Federal de Sao Paulo.; Brasi

Cytotoxic effects of essential oils from three Lippia gracilis Schauer genotypes on HeLa, B16, and MCF-7 cells and normal human fibroblasts

This study aimed to evaluate the chemical composition of the essential oils from three genotypes of Lippia gracilis Schauer (Verbenaceae) and investigate the cytotoxic activities of these oils. Essential oils were extracted from the leaves using a Clevenger-type apparatus, and chemical analysis was performed using a gas chromatograph coupled to a mass spectrometer and flame ionization detector. 3T3, MRC5, B16, HeLa, and MCF-7 cell lines were used to study the in vitro cytotoxicity of the essential oils, and the level of cell death was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test with three replicates. The cytotoxic activity was expressed as the concentration that inhibited 50% of cell growth. The main compound in the essential oil of LGRA-106 was thymol (40.52%), while LGRA-109 and LGRA-201 contained 45.84 and 32.60% carvacrol, respectively, as their major compound. The essential oils of L. gracilis showed cytotoxic activity against both normal and tumor cells at concentrations below 100 μg/mL; this demonstrated the antitumor potential of these essential oils, which should be further investigated