Dynamized Aloysia Polystachya (Griseb.) Essential Oil: A Promising Antimicrobial Product.

BACKGROUND: Compounds from vegetal matter have therapeutic potential to control highly prevalent microorganisms that are resistant to commonly used antimicrobial drugs. Dynamization of compounds can either maintain or improve their therapeutic effects, and make their use safer, especially those compounds whose therapeutic dose is close to the toxic limit. Aloysia polystachya (Griseb.) stands out among aromatic plants with antimicrobial potential. OBJECTIVE: The aim of this study was to evaluate the antimicrobial activity of dynamized and crude forms of A. polystachya essential oil against Candida albicans, Escherichia coli and Staphylococcus aureus. METHODS: Essential oil was extracted from A. polystachya dry leaves, solubilized, and dynamized at 1 cH potency as recommended by the Brazilian Homeopathic Pharmacopoeia. Antimicrobial activity against C. albicans, E. coli and S. aureus of the samples was assayed using the plate microdilution method. RESULTS: Dynamized A. polystachya essential oil at the concentration of 1 µg/mL inhibited the growth of all the microbial species analyzed. The minimum inhibitory concentration of dynamized essential oil was smaller than crude essential oil for S. aureus, E. coli and C. albicans. CONCLUSION: It is reported for the first time that A. polystachya dynamized essential oil can effectively suppress microbial growth, and it is a promising adjuvant to treat infections with pathogenic S. aureus, E. coli and C. albicans.

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

Abstract 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.

Microtranscriptome analysis of sugarcane cultivars in response to aluminum stress.

Although several metallic elements are required for plant growth, excessive amounts of aluminum ions (Al3+) can result in the inhibition of root growth, thus triggering water and nutrient deficiencies. Plants under stress undergo gene expression changes in specific genes or post-transcriptional gene regulators, such as miRNAs, that can lead to stress tolerance. In this study, we investigated the miRNAs involved in the response of sugarcane to aluminum stress. Four miRNA libraries were generated using sugarcane roots of one tolerant and one sensitive sugarcane cultivar grown under aluminum stress and used to identify the miRNAs involved in the sugarcane aluminum toxicity response. The contrast in field phenotypes of sugarcane cultivars in the field during aluminum stress was reflected in the micro-transcriptome expression profiles. We identified 394 differentially expressed miRNAs in both cultivars, 104 of which were tolerant cultivar-specific, 116 were sensitive cultivar-specific, and 87 of which were common among cultivars. In addition, 52% of differentially expressed miRNAs were upregulated in the tolerant cultivar while the majority of differentially expressed miRNAs in the sensitive cultivar were downregulated. Real-time quantitative polymerase chain reaction was used to validate the expression levels of differentially expressed miRNAs. We also attempted to identify target genes of miRNAs of interest. Our results show that selected differentially expressed miRNAs of aluminum-stressed sugarcane cultivars play roles in signaling, root development, and lateral root formation. These genes thus may be important for aluminum tolerance in sugarcane and could be used in breeding programs to develop tolerant cultivars.

Chalcone Derivatives 4'-Amino-1-Naphthyl-Chalcone (D14) and 4'-Amino-4-Methyl-1-Naphthyl-Chalcone (D15) Suppress Migration and Invasion of Osteosarcoma Cells Mediated by p53 Regulating EMT-Related Genes.

Osteosarcoma (OS) is a primary malignant bone tumor that mainly affects children, adolescents, and young adults. The inhibition of metastasis is a main strategy of OS therapy since the development of metastatic disease due to drug resistance remains the most important cause of death from this cancer. Considering the severe side effects of current OS chemotherapy, the identification of anti-metastatic drugs with reduced toxicity is of great interest. Chalcones are polyphenols with a basic structure consisting of an α-, ß-unsaturated carbonyl system linking two aryl rings. These compounds exhibit anticancer activity against a variety of tumor cell lines through multiple mechanisms, including the regulation of the tumor-suppressor protein p53 and its target genes. An important process regulated by p53 is epithelial-mesenchymal transition (EMT), which facilitates tumor metastasis by conferring migratory and invasive properties to cancer cells. The activation of p53 can revert EMT and reduce migration and invasion. This study aimed to examine the inhibitory effects of two 4'-aminochalcones on the migration/invasion of the U2OS (p53+/+) and SAOS-2 (p53-/-) OS cell lines as well as the underlying molecular mechanisms. Cell viability was examined by MTT assay. Transwell assays were used to evaluate the migratory and invasive ability of the cells. The two 4'-aminochalcones showed low capacity to inhibit the viability of OS cells independent of p53 status, but preferentially suppressed the migration of U2OS cells and of a SAOS-2 cell line expressing p53. Invasion was strongly inhibited by both chalcones independent of p53 status. RT-PCR, zymography, and Western blot were used to study the expression of matrix metalloproteinases and EMT markers after treatment with the chalcones. The results indicated that the 4'-aminochalcone-induced antimigratory and anti-invasive effects are potentially associated with the inhibition of extracellular matrix (ECM) enzymatic degradation in OS cells and with the modulation of EMT genes. These effects probably result from the induced increase of p53 protein expression by the two chalcones. In conclusion, chalcones D14 and D15 have potential anti-metastatic activity mediated by p53 that can be exploited for OS treatment.

Genetic and chemical diversity of Uncaria tomentosa (Willd. ex. Schult.) DC. in the Brazilian Amazon.

Uncaria tomentosa (Willd. ex Schult.) DC., a plant native to the Amazon region, is used widely in popular medicine and by the pharmaceutical industry because of its anti-inflammatory activity. However, the survival of this species is endangered by deforestation and indiscriminate collection, and a preservation plan is urgently required. The objectives of this study were to determine the genetic and chemical variability between and within eight populations of U. tomentosa from the Brazilian states of Acre, Pará and Amapá, and to investigate possible correlations between genetic and geographical distances, and between geographical distances or altitude and the accumulation of bioactive oxindole alkaloids. Three sequence-related amplified polymorphism (SRAP) markers were employed to fingerprint genomic DNA, and the amounts of mitraphylline and isomitraphylline in leaf samples were established by high-performance liquid chromatography. Although significant divergence existed between the tested populations (FST = 0.246), the largest genetic diversity and the highest percentage of polymorphism (95.68%) was found within the population from Mâncio Lima, Acre. Gene flow was considered rather limited (Nm = 1.57), and no correlations between genetic and geographical distances were detected, suggesting that population structure followed an island model. Accumulations of mitraphylline and isomitraphylline varied in the range 32.94 to 0.57 and 3.75 to 0.36 mg g-1 dry weight, respectively. The concentration of isomitraphylline was positively influenced by altitude, such that the population collected at the site with the highest elevation (Tarauacá, Acre) exhibited the greatest alkaloid content. SRAP markers were very efficient in fingerprinting genomic DNA from U. tomentosa populations and clearly showed that genetic variability within populations was greater than between populations. A conservation and management plan should prioritize the creation of germplasm banks to prevent the loss of existing genetic variability, particularly within alkaloid-rich populations such as those of Tarauacá.

Biosurfactants produced by Microbacterium sp, isolated from aquatic macrophytes in hydrocarbon-contaminated area in the Rio Negro, Manaus, Amazonas / Biossurfactantes produzidos por Microbacterium sp, isolada de macrófita aquática em área impactada por hidrocarbonetos no rio Negro, Manaus, Amazonas

Endophytic bacteria isolated from Eichhornia crassipes (Mart) Solms., collected in oil contaminated wastewater of effluent generated by Petrobras refinery in Manaus were investigated to determine their potential for producing biosurfactants. Assay with 2.6-dichlorophenol indophenol (DCPIP) indicator to verify hydrocarbon biodegradation activity; oil emulsification test; drop-collapse method; surface tension and growth curve of biosurfactant production. The M87 Microbacterium sp. strain chosen for this work was identified by the sequencing of the rDNA region and the chemical characterization was performed by FTIR, UFLC/MS and 1H RMN techniques. The selected bacterial isolate provided 3g L-1 of biosurfactant, using diesel oil as sole carbon source, being efficient in biodegrading oil as demonstrated by the DCPIP test. Fractions obtained by column chromatography were efficient in reducing water surface tension around 40 mN m-1, especially fraction 1, which reduced it to 34.17 mN m-1. The different techniques of chemical analysis used for the identification of the biosurfactant isolate indicated that this is probably a long - chain fatty acid lipid type, which may be used in the future as both biosurfactant in decontamination processes of hydrocarbon-polluted areas or as bioemulsifier in countless processes, since it exhibited no toxicity as determined by Alamar Blue assay.

Foram investigadas bactérias endofíticas isoladas de Eichhornia crassipes (Mart.) Solms., coletadas em águas contaminadas com resíduos de petróleo em um afluente da refinaria da Petrobrás/Manaus, para avaliação da produção de biossurfactantes. Para selecionar o micro-organismos à produção e caracterização de biossurfactantes, foram realizados os seguintes testes: a descoloração do indicador 2,6 indofenol (DCPIP), emulsificação do diesel, colapso da gota, tensão superficial e curva de produção. A caracterização química foi realizada por meio das técnicas de FT-IR, UFLC/MS e RMN1H. A bactéria M87 Microbacterium neste estudo, foi identificada pelo sequenciamento da região rDNA e produziu 3g L-1 de biossurfactantes utilizando o diesel como fonte de carbono, mostrando-se eficiente na ação biodegradadora do petróleo, por meio do teste de Indofenol (DCPIP). As frações obtidas, mostraram-se eficazes na redução da tensão superficial da água abaixo de 40 mN m-1, com destaque para a fração 1 que reduziu a tensão superficial para 34,17 mN m-1. Pelas análises química utilizadas, pode-se inferir que, provavelmente, se trata de um ácido graxo de cadeia longa, que pode ser utilizado futuramente tanto como biossurfactante em processos de descontaminação de ambientes impactados por hidrocarbonetos, assim como bioemulsificante em inúmeros processos uma vez que não apresentou toxicidade por meio do teste realizado.

A standardized methanol extract of Eclipta prostrata (L.) L. (Asteraceae) reduces bronchial hyperresponsiveness and production of Th2 cytokines in a murine model of asthma.

ETHNOPHARMACOLOGICAL RELEVANCE: Eclipta prostrata (L.) L. (Asteraceae) has been used in Brazilian traditional medicine to treat asthma and other respiratory illnesses. AIMS OF THE STUDY: To investigate the effects of different doses of a standardized extract of E. prostrata using a murine model of allergen induced asthma. MATERIALS AND METHODS: Balb/c mice were sensitized twice with ovalbumin (OVA) administered intraperitoneally and challenged over four alternate days with nasal instillations of OVA solution. The standardized methanol extract of E. prostrata was administered in doses of 100, 250 and 500mgkg-1 concomitantly with nasal instillation over seven consecutive days. Control animals were treated with dexamethasone or saline solution. Bronchial hyperresponsiveness, production of Th1 and Th2 cytokines, allergen sensitization, airway and lung inflammation, mucous secretion and airway remodeling were assessed. RESULTS: The concentrations of chemical markers in the standardized methanol extract were 0.02% oroboside, 1.69% demethylwedelolactone and 1.71% wedelolactone. Treatment with 250mgkg-1 of extract, which provided 0.745, 4.22 and 4.30mgkg-1day-1 of oroboside, demethylwedelolactone and wedelolactone, respectively, significantly reduced (P<0.05) respiratory resistance and elastance. Such effects were comparable with those produced by dexamethasone. The total number of inflammatory cells and eosinophils in the bronchoalveolar lavage and the concentrations of interleukin (IL)-4, IL-5 and IL-13 in lung homogenate were significantly reduced (P<0.05) by the methanol extract of E. prostrata. CONCLUSION: The results presented herein demonstrate for the first time the anti-inflammatory activity of E. prostrata in a murine model of asthma, thereby supporting the ethnopharmacological uses of the plant.

The glycosylated flavonoids vitexin, isovitexin, and quercetrin isolated from Serjania erecta Radlk (Sapindaceae) leaves protect PC12 cells against amyloid-ß25-35 peptide-induced toxicity.

The Aß peptide-mediated toxicity participates in the neuronal death that occurs in Alzheimer's disease. The present study aims to isolate the major compounds of Serjania erecta Radlk leaves and assess whether these compounds protect PC12 cells from Aß25-35 peptide-induced toxicity. We isolated three flavonoid glycosides with high purity: quercetrin, vitexin, and isovitexin. The Aß25-35 peptide alone decreased the PC12 cell viability in a concentration-dependent manner, as evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. We selected the Aß25-35 peptide concentration of 50 µM for the experiments. Treatment of PC12 cells with the flavonoids before exposure to the Aß25-35 peptide increased cell viability, i.e., these compounds protected the cells against Aß25-35 peptide-induced toxicity. Vitexin promoted higher protection levels than quercetrin and isovitexin, and reduced the lactate dehydrogenase release and NO production in Aß25-35 peptide-treated PC12 cells. Therefore, the glycosylated flavonoids that exist in S. erecta leaves, especially vitexin, protect PC12 cells from Aß25-35 peptide-induced toxicity.

Nematocidal effects of natural phytoregulators jasmonic acid and methyl-jasmonate against Pratylenchus zeae and Helicotylenchus spp.

The aim of this study was to evaluate the nematocidal effects of natural phytoregulators jasmonic acid (JA) and methyl-jasmonate (MJ) against plant parasitic nematodes Pratylenchus zeae (Graham) (Pratylenchidae) and Helicotylenchus spp. (Hoplolaimidae). Both JA and MJ promoted elevated percentages of mortality in P. zeae and Helicotylenchus spp. after 12 and 24 h of nematodes exposition at different concentrations of jasmonates. Considering the potential use of jasmonates as biofertiliser added now for their nematocidal effects, our results are of relevance in terms of biotechnological application.

Micropropagação de Croton antisyphiliticus Mart. / Micropropagation of Croton antisyphiliticus Mart.

Croton antisyphiliticus Mart. ex M. Arg., conhecido popularmente como pé-de-perdiz, é uma planta medicinal nativa do Cerrado, cuja raiz é utilizada na forma de decoctos para combater infecções do aparelho reprodutor masculino e feminino. A coleta da planta é realizada de forma extrativista e não há trabalhos a respeito da sua conservação. O objeto do trabalho foi estabelecer um protocolo de micropropagação de Croton antisyphiliticus. Foram avaliados o efeito de diferentes concentrações das citocininas, o tamanho do recipiente para o cultivo in vitro, o co-cultivo e a posição da gema em relação ao desenvolvimento dos explantes, bem como a influência do AIB no enraizamento in vitro e a aclimatização dos explantes. O melhor desenvolvimento in vitro foi obtido com explantes do tipo gema apical, co-cultivadas em meio MS suplementado com BAP (1µM), em frasco tipo pote. O enraizamento foi obtido em meio MS sem regulador vegetal e a aclimatização pode ser realizada em solo do cerrado com brotações sem a presença de raízes desenvolvidas in vitro.

Croton antisyphiliticus Mart. ex M. Arg., popularly known as "pé-de-perdiz", is a native medicinal plant of Cerrado, whose root is used in the form of decoctions against infections of the male and female reproductive. The collection of the plant is carried through of extractives form and there are no studies regarding its conservation. The aim of this study was to establish a micropropagation protocol of C. antisyphiliticus. The effect of different concentrations of cytokinins, the size of the container for the in vitro culture, co-cultivation and bud position in relation to the development of the explants, as well as the influence of AIB on in vitro rooting and acclimatization of explants were evaluated. The best development in vitro was obtained with apical explants co-cultivated on MS medium supplemented with BAP (1µM) in bottle type pot. Rooting was achieved on MS medium without growth regulators and acclimatization can be performed in Cerrado soil without the presence of shoots developed roots in vitro.

Micropropagação e conservação de Macrosyphonia velame (St. Hil.) Muell. Arg. em banco de germoplasma in vitro / Micropropagation and conservation of Macrosyphonia velame (St. Hil.) Muell. Arg. in vitro germoplasm bank

A Macrosyphonia velame é uma planta medicinal do Cerrado, pertencente à família Apocynaceae, e conhecida popularmente como velame branco. Extratos de raízes de velame são utilizados pela população como depurativo e anti-sifilítico. Este trabalho teve como objetivo desenvolver o protocolo de micropropagação de M. velame, com vistas à conservação da espécie em Banco do Germoplasma in vitro. Sementes foram coletadas nos municípios de Sacramento, Tapira e Araxá, MG, e utilizadas como fonte de explantes. Segmentos nodais de plântulas axênicas foram transferidos para meio MS/2 suplementado com BAP, Cinetina, 2ip e TDZ em diferentes concentrações (0,25; 5,0 e 1,0mg L-1). Para o enraizamento, brotações com 2cm de altura foram transferidas para meios MS/2 suplementado com 1,0; 5,0; e 10,0mg L-1 de IBA ou ANA e cultivadas por 5, 10 e 30 dias, sendo em seguida subcultivadas em MS/2 por mais trinta dias e só então avaliadas quanto à formação de raízes. Na aclimatização e enraizamento ex vitro, foram utilizados substratos Plantmax®, areia e solo individualmente ou em combinações (1:1) de areia/Plantmax®; areia/solo; Plantmax®/solo. Para o estabelecimento do Banco de Germoplasma, brotações com 3,5cm foram transferidas para meio MS/2 suplementado com 2 por cento de sacarose + 4 por cento de manitol ou sorbitol; 2 por cento de sacarose + 4 por cento de manitol ou sorbitol + 2mg L-1 de pantotenato de cálcio; 2 por cento de sacarose + 4 por cento de manitol ou sorbitol + 2mg L-1 de espermidina. A porcentagem de germinação in vitro foi baixa, 33 por cento, 4 por cento e 2 por cento das sementes coletadas em Araxá, Tapira e Sacramento, respectivamente. O meio MS/2 sem adição de citocinina promoveu a proliferação de brotos (4,0 por gema), elongação (5,2cm), número de gemas (8,6) e reduziu vitrificação (4 por cento). Em relação aos substratos testados, as plântulas se desenvolveram melhor no Plantmax®, sendo que 40 por cento das plântulas sobreviveram e a maioria apresentou formação de raízes. Plântulas cultivadas por três meses em meio de cultura MS/2 + 2 por cento de sacarose + 4 por cento de manitol + 2mg L-1 de pantotenato de cálcio, sob condições de Banco de Germoplasma, apresentaram 40 por cento de sobrevivência.

Root extracts of Macrosyphonia velame an Apocynaceae native of Brazilian Cerrado, known as white velame have been popularly used as depurative and anti-syphilitic agent. The aim of the present research was to develop a micropropagation protocol for the in vitro conservation of M. velame in a germplasm bank. Seeds of velame collected in Sacramento, Tapira and Araxá, MG, Brazil, were used as initial explants. Nodal segments from axenic plantlets were inoculated on MS/2 medium supplemented with different concentrations (0.25, 5.0 and 1.0mg L-1) of BAP; kinetin; 2iP or TDZ. For in vitro rooting , plantlets (2cm high) were inoculated on MS/2 medium supplemented with IBA or NAA (1.0, 5.0, and 10.0mg L-1), maintained for 5, 10 and 30 days and sub-cultured to MS/2 medium for an additional thirty days before evaluating rooting. For acclimatization and ex vitro rooting plantlets were transplanted into Styrofoam boxes containing either Plantmax®, sand and soil one by one or in combinations (1:1) of sand/Plantmax®; sand/soil; Plantmax®/soil. For the in vitro conservation of M. velame in germplasm bank plantlets (3.5cm high) were inoculated on MS/2 medium supplemented with either 2 percent sucrose + 4 percent of mannitol or sorbitol; 2 percent sucrose + 4 percent mannitol or sorbitol + 2mg L-1 calcium pantothenate; 2 percent of sucrose + 4 percent of mannitol or sorbitol + 2mg L-1 spermidine. The proportion of seed germination was considered low, 33 percent, 4 percent and 2 percent for seeds collected in Araxá, Tapira and Sacramento respectively. Explants cultured on MS/2 medium without addition of cytokinin showed enhanced height (5.2cm), increased number of buds (8.6), proliferation of 4 shoots per bud and minimal (4 percent) proportion of vitrification. Plantlets acclimatized ex vitro developed better in Plantmax® substrate, most plantlets presented root formation and survival reached 40 percent. M. velame plantlets cultured for three months on MS/2 added with 2 percent sucrose + 4 percent mannitol + 2mg L-1 of calcium pantothenate, under germoplasma bank conditions presented 40 percent of survival.

Micropropagação de Dioscorea multiflora Grised / Micropropagation of Dioscorea multiflora Grised

Dioscorea multiflora uma planta nativa do Sul do Brasil produz a diosgenina como metabólito secundário majoritário, uma substância potencialmente usada pela indústria farmacêutica para a produção de cortisona e substâncias com ação contraceptiva. Objetivou-se, neste trabalho otimizar o protocolo de micropropagação de D. multiflora, visando a produção de mudas em escala comercial. Segmentos nodais subcultivados em meio MS sólido foram transferidos para multiplicação em meio MS suplementado com BAP (0,01; 0,1; 0,5; 1,0 e 3,0 mg L-1)e meio MS suplementado com 0,1 mg L-1 ou 0,5 mg L-1 de BAP acrescido de diferentes concentrações de sacarose (2, 4, 6, 8 e 10 por cento). Para o enraizamento, as brotações foram cultivadas em meio MS suplementado com AIB (0,1; 0,5; 1,0 e 3,0 mg L-1) e meio MS suplementado com ANA (0,1; 0,5; 1,0 e 3,0 mg L-1). Os experimentos in vitro foram instalados em delineamento experimental inteiramente casualizado e cada tratamento constituiu-se de 3 repetições e 10 cubetas/parcela. Plântulas com e sem raízes foram aclimatizadas em casa de vegetação. Melhores resultados de multiplicação e enraizamento foram obtidos em meio MS + 0,1 mg L-1 de BAP (80 por cento) e em meio MS + 1,0 mg L-1 de AIB (42,6 por cento), respectivamente. Não houve diferença quanto à porcentagem de sobrevivência das plântulas in vitro e ex vitro durante a aclimatização (75 por cento). O protocolo de micropropagação para D. Multiflora é efetivo e pode ser usado para a produção em escala comercial.

Dioscorea multiflora is a plant native to southern Brazil that produces diosgenin as a major secondary metabolite, a substance which is used by the pharmaceutical industry for the production of cortisone and substances with contraceptive action. The objective of this work was to optimize the micropropagation protocol of D. multiflora, for the production of seedlings on a commercial scale. Nodal segments subcultured in solid MS medium were transferred for multiplication to MS medium supplemented with BAP (0.01, 0.1, 0.5, 1.0 and 3.0 mg L-1) and MS medium supplemented with 0.1 mg L-1 or 0.5 mg L-1 BAP plus different concentrations of sucrose (2, 4, 6, 8 and 10 percent). For rooting, the shoots were cultured on MS medium supplemented with IBA (0.1, 0.5, 1.0 and 3.0 mg L-1) and MS medium supplemented with NAA (0.1, 0.5, 1.0 and 3.0 mg L-1). A completely randomized design was used with treatment consisting of 3 replicates with 10 buckets per plot. Seedlings with and without roots were acclimatized in a greenhouse. The best results of multiplication and rooting were obtained in MS medium + 0.1 mg L-1 BAP (80 percent) and in MS medium + 1.0 mg L-1 IBA (42.6 percent), respectively. There was no difference in the survival percentage of seedlings in vitro and during ex vitro acclimatization (75 percent). The micropropagation protocol for production of D. multiflora is effective and can be used for commercial production.

Serotonin receptor (5-HT 2A) and catechol-O-methyltransferase (COMT) gene polymorphisms: triggers of fibromyalgia?

INTRODUCTION: Fibromyalgia is a rheumatic syndrome characterized by diffuse and chronic pain associated with fatigue, sleep disorders, anxiety, depression, memory loss, and dizziness. Although the physiological mechanisms that control fibromyalgia have not been precisely established, neuroendocrine, genetic or molecular factors may be involved in fibromyalgia. OBJECTIVE: The aim of the present study was to characterize serotonin receptor (5-HT2A) and catechol-O-methyltransferase (COMT) gene polymorphisms in Brazilian patients with fibromyalgia and to evaluate the participation of these polymorphisms in the etiology of the disease. MATERIAL AND METHODS: Genomic DNA extracted from 102 blood samples (51 patients, 51 controls) was used for molecular characterization of the 5-HT2A and COMT gene polymorphisms by PCR-RFLP. RESULTS: Analysis of the 5-HT2A polymorphism revealed a frequency of 25.49% C/C, 49.02% T/C and 25.49% T/T in patients, and of 17.65% C/C, 62.74% T/C and 19.61% T/T in the control group, with no differences between the two groups.Analysis of the COMT polymorphism in patients showed a frequency of 17.65% and 45.10% for genotypes H/H and L/H, respectively. In the control group the frequency was 29.42% for H/H and 60.78% for L/H, also with no differences between the two groups. However, there was a significant difference in the frequency of the L/L genotype between patients (37.25%) and controls (9.8%), which permitted differentiation between the two groups. CONCLUSION: The L/L genotype was more frequent among fibromyalgia patients. Though considering a polygenic situation and environmental factors, the molecular study of the rs4680 SNP of the COMT gene may be helpful to the identification of susceptible individuals.

Germinação de sementes in vitro e desenvolvimento de plantas ex vitro de algodãozinho-do-campo / in vitro seeds germination and ex vitro plants development of 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 por cento) 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 percent 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.

Polimorfismos dos genes do receptor de serotonina (5-HT2A) e da catecol-O-metiltransferase (COMT): fatores desencadeantes da fibromialgia? / Serotonin receptor (5-HT 2A) and catechol-O-methyltransferase (COMT) gene polymorphisms: Triggers of fibromyalgia?

INTRODUÇÃO: A fibromialgia é uma síndrome reumática caracterizada por dor difusa e crônica associada a fadiga, insônia, ansiedade, depressão, perda de memória e tontura. Embora os mecanismos fisiológicos que controlam a fibromialgia não tenham sido estabelecidos, fatores neuroendócrinos, genéticos ou moleculares podem estar envolvidos. OBJETIVO: O objetivo do presente estudo foi caracterizar os polimorfismos dos genes do receptor de serotonina (5-HT2A) e da catecolO-metiltransferase (COMT) em pacientes brasileiros com fibromialgia, a fim de avaliar sua participação na etiologia da doença. MATERIAL E MÉTODOS: O DNA genômico extraído de 102 amostras de sangue (51 pacientes, 51 controles) foi usado para a caracterização molecular dos polimorfismos dos genes 5-HT2A e COMT, por meio de PCR-RFLP. RESULTADOS: A análise molecular dos polimorfismos do gene 5-HT2A demonstrou frequências de 25,49 por cento C/C, 49,02 por cento T/C e 25,49 por cento T/T, nos pacientes com fibromialgia, e 17,65 por cento C/C, 62,74 por cento T/C e 19,61 por cento T/T, no grupo controle, não apresentando diferença significativa entre o grupo de pacientes e o grupo controle. Os polimorfismos do gene da COMT em pacientes com fibromialgia apresentaram uma frequência de 17,65 por cento e 45,10 por cento para os genótipos H/H e L/H, respectivamente. No grupo controle, as frequências foram de 29,42 por cento, para H/H, e 60,78 por cento, para L/H, sem diferença significativa entre ambos os grupos. Entretanto, houve diferença significativa na frequência do genótipo L/L em pacientes (37,25 por cento) e controles (9,8 por cento), o que permitiu a diferenciação entre os dois grupos. CONCLUSÃO: A frequência do genótipo L/L foi maior nos pacientes com fibromialgia. Apesar de a fibromialgia envolver uma situação poligênica e fatores ambientais, o estudo molecular do SNP rs4680 do gene da COMT pode auxiliar a identificação de indivíduos suscetíveis.

INTRODUCTION: Fibromyalgia is a rheumatic syndrome characterized by diffuse and chronic pain associated with fatigue, sleep disorders, anxiety, depression, memory loss, and dizziness. Although the physiological mechanisms that control fibromyalgia have not been precisely established, neuroendocrine, genetic or molecular factors may be involved in fibromyalgia. OBJECTIVE: The aim of the present study was to characterize serotonin receptor (5-HT2A) and catecholO-methyltransferase (COMT) gene polymorphisms in Brazilian patients with fibromyalgia and to evaluate the participation of these polymorphisms in the etiology of the disease. MATERIAL AND METHODS: Genomic DNA extracted from 102 blood samples (51 patients, 51 controls) was used for molecular characterization of the 5-HT2A and COMT gene polymorphisms by PCR-RFLP. RESULTS: Analysis of the 5-HT2A polymorphism revealed a frequency of 25.49 percent C/C, 49.02 percent T/C and 25.49 percent T/T in patients, and of 17.65 percent C/C, 62.74 percent T/C and 19.61 percent T/T in the control group, with no differences between the two groups.Analysis of the COMT polymorphism in patients showed a frequency of 17.65 percent and 45.10 percent for genotypes H/H and L/H, respectively. In the control group the frequency was 29.42 percent for H/H and 60.78 percent for L/H, also with no differences between the two groups. However, there was a significant difference in the frequency of the L/L genotype between patients (37.25 percent) and controls (9.8 percent), which permitted differentiation between the two groups. CONCLUSION: The L/L genotype was more frequent among fibromyalgia patients. Though considering a polygenic situation and environmental factors, the molecular study of the rs4680 SNP of the COMT gene may be helpful to the identification of susceptible individuals.

Atividade antioxidante de Jacaranda decurrens Cham., Bignoniaceae / Antioxidant activity of Jacaranda decurrens Cham., Bignoniaceae

Muitas doenças e processos degenerativos estão associados à superprodução de espécies reativas de oxigênio (ERO) o que tem estimulado vários grupos de pesquisa a investigarem o potencial antioxidante de substâncias produzidas por diversas famílias da flora mundial. O presente trabalho teve como objetivo avaliar a atividade antioxidante de extrato bruto hidroalcoólico de folhas de Jacaranda decurrrens e de frações obtidas a partir desse extrato, pelo método espectrométrico de descoloração do radical 2,2-difenil-1-picrilhidrazila (DPPH). A atividade antioxidante (91 por cento) do extrato bruto e da fração Jd-2 foi semelhante à atividade obtida com o padrão rutina, na concentração de 1 mg/L (92,56 por cento). A fração Jd-3 quando comparada com as outras frações apresentou maior atividade antioxidante em concentrações igual ou abaixo de 2,5 mg/L. A atividade antioxidante das amostras está relacionada à presença dos triterpenos ácidos ursólico e oleanólico presentes no extrato bruto e nas frações Jd-1 e Jd-2 e também a flavonóides glicosilados contidos na fração Jd-3. Este é o primeiro trabalho demonstrando a atividade antioxidante de extrato de folhas de Jacaranda decurrens.

Since many diseases and degenerative processes have been associated to the overproduction of reactive oxygen species (ROS) many research groups are motivated to investigate the antioxidant potential of substances produced by several families of the world flora. The aim of this work was to evaluate the antioxidant activity of crude hydroalcoholic extract of Jacaranda decurrrens leaves and fractions obtained from that extract, using the free radical DPPH (2,2-difenyl-1-picrylhydrazyl) through spectrometric discoloration method. Results confirmed that either the crude extract or fractions of the extract presented antioxidant activity. The antioxidant activity (91 percent) demonstrated by the crude extract was equivalent to the antioxidant activity determined for fraction Jd-2 in the concentration of 10 mg/L, and similar to the activity showed by the standard rutin, in the concentration of 1 mg/L (92,56 percent). Fraction Jd-1 in the concentration of 10 mg/L and 5 mg showed 84 percent and 86 percent antioxidant activity, respectively. Those values are significantly different and inferior if compared to the standard rutin. Compared to the other fractions the Jd-3 presented higher antioxidant activity. The antioxidant potential of Jacaranda decurrens crude extracts and fractions Jd-1 and Jd-2 is probably related to the production of the triterpenes ursolic and oleanolic acids and also to the glycosylated flavonoids produced in the fraction Jd-3. This is the first report on the antioxidant activity of crude hydroalcoholic extract of J. decurrrens leaves and fractions obtained from that extract.

Antimicrobial activity of Arctium lappa constituents against microorganisms commonly found in endodontic infections

A atividade antimicrobiana de extratos brutos de folhas de Arctium lappa, bem como de suas fases, foi avaliada in vitro. Os microrganismos Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis e Candida albicans, comuns na cavidade bucal, especificamente em infecções endodônticas, foram utilizados. O método de difusão em Agar permitiu a detecção da fase hexânica como inibitória do crescimento microbiano. Ensaios de bioautografia identificaram substâncias antimicrobianas presentes no extrato. Os resultados demonstraram a presença, na fase hexânica bruta e em suas sub-frações, de constituintes que têm Rf (fatores de retenção) em três zonas distintas, sugerindo a presença de ativos com estruturas químicas de diferentes polaridades, que exibiram especificidade contra os microrganismos alvos. Conclui-se que os constituintes de Arctium lappa apresentam um grande potencial de inibição microbiana contra os microrganismos endodônticos estudados.

Isolation and structure determination of bioactive isoflavones from callus culture of Dipteryx odorata.

Crude extracts of callus and roots of Dipteryx odorata were analyzed by HPLC to detect and quantify isoflavone contents. Based on spectroscopic and X-ray crystallography data the structures of two isoflavones were elucidated as 7-hydroxy-4',6-dimethoxyisoflavone and 3',7-dihydroxy-4',6-dimethoxyisoflavone. The production of dry biomass of 7-hydroxy-4',6-dimethoxyisoflavone in cultured callus was 4.12 mg/g, approximately eleven fold higher than the amount accumulated in roots of D. odorata wild-growing plants. The 7-hydroxy-4',6-dimethoxyisoflavone was effective against glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi. This is the first report on those bioactive isoflavones accumulated in callus of D. odorata.

Antimicrobial activity of Arctium lappa constituents against microorganisms commonly found in endodontic infections.

This study evaluated in vitro the antimicrobial activity of rough extracts from leaves of Arctium lappa and their phases. The following microorganisms, commonly found in the oral cavity, specifically in endodontic infections, were used: Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis and Candida albicans. The agar-diffusion method allowed detection of the hexanic phase as an inhibitor of microbial growth. Bioautographic assays identified antimicrobial substances in the extract. The results showed the existence, in the rough hexanic phase and in its fractions, of constituents that have retention factors (Rf) in three distinct zones, thereby suggesting the presence of active constituents with chemical structures of different polarities that exhibited specificity against the target microorganisms. It may be concluded that the Arctium lappa constituents exhibited a great microbial inhibition potential against the tested endodontic pathogens.

Antimicrobial screening and quantitative determination of benzoic acid derivative of Gomphrena celosioides by TLC-densitometry.

The antimicrobial activity of ethanolic extract and pure compounds of Gomphrena celosioides have been screened by Kirby-Bauer method. Quantitative determination of 4-hydroxy-3-methoxy-benzoic acid in stems, leaves, flowers and roots was established by TLC-densitometry. Results showed significant activity against Staphylococcus aureus and Salmonella typhi. There were no significant differences in the determined benzoic acid derivative.