Authentication of Brazilian Ginseng using Bar-HRM analysis

Abstract Hebanthe eriantha (Martius) Kuntze and Pfaffia glomerata (Spreng) Pedersen are medicinal plants popularly known as "Brazilian Ginseng" due to their similarity to Panax ginseng. In Brazil, they are sold as the same herb, despite their different pharmacological and toxicological properties. The morphological identification is difficult, which facilitates their adulteration. We report the application of the Barcode DNA High-Resolution Melting (Bar-HRM) using matK gene to differentiate both species in samples sold in the Brazilian market. Using the proposed method, we could discriminate and identify both species. Bar-HRM analysis allowed discriminating and identifying both species. It allowed the identification of H. eriantha and P. glomerata in 43.6% and 56.4% of the amplified samples, respectively. Of these, only seven samples were authenticated and, in 71.4% of the cases, adulterated. We concluded that Bar-HRM has proven to be a fast alternative method to authenticate plants under the common name "Brazilian Ginseng".

Biological properties of selected Amaranthaceae halophytic species: A review

Abstract The family Amaranthaceae contains a large number of halophytic species. The aim of this paper is to present a review of biologically significant halophytes in the flora of Serbia belonging to this family. This review includes a description of the following 18 species: Atriplex littoralis L., A. rosea L., A. halimus L., Bassia sedoides Pall., B. scoparia (L.) A.J.Scott., B. prostrata (L.) Beck., Beta trigyna W. et K., Camphorosma annua Pall., C. monspeliaca L., Chenopodium ambrosioides L., Ch. polyspermum L., Ch. vulvaria L., Ch. hybridum L., Ch. album L., Ch. rubrum L., Salicornia europaea L., Suaeda maritima (L.) Dum., Salsola soda L. These species are sources of biologically active substances and have a good potential for multi-purpose applications. Most extracts of these species have been found to exhibit biological activities such as antioxidant, anticancer, antibacterial, antifungal, antiinflammatory and others. The aim of this review is to describe these species including their morphology, distribution, phytochemistry, as well as their use for medicinal and food purposes.

In vitro antimicrobial and antimycobacterial activity and HPLC-DAD screening of phenolics from Chenopodium ambrosioides L

Abstract The main objective of this study was to demonstrate the antimicrobial potential of the crude extract and fractions of Chenopodium ambrosioides L., popularly known as Santa-Maria herb, against microorganisms of clinical interest by the microdilution technique, and also to show the chromatographic profile of the phenolic compounds in the species. The Phytochemical screening revealed the presence of cardiotonic, anthraquinone, alkaloids, tannins and flavonoids. The analysis by HPLC-DAD revealed the presence of rutin in the crude extract (12.5 ± 0.20 mg/g), ethyl acetate (16.5 ± 0.37 mg/g) and n-butanol (8.85 ± 0.11 mg/g), whereas quercetin and chrysin were quantified in chloroform fraction (1.95 ± 0.04 and 1.04 ± 0.01 mg/g), respectively. The most promising results were obtained with the ethyl acetate fraction, which inhibited a greater number of microorganisms and presented the lowest values of MIC against Staphylococcus aureus and Enterococcus faecalis (MIC = 0.42 mg/mL), Pseudomonas aeruginosa (MIC = 34.37 mg/mL), Paenibacillus apiarus (MIC = 4.29 mg/mL) and Paenibacillus thiaminolyticus (MIC = 4.29 mg/mL). Considering mycobacterial inhibition, the best results were obtained by chloroform fraction against M. tuberculosis, M. smegmatis, and M. avium (MIC ranging from 156.25 to 625 µg/mL). This study proves, in part, that the popular use of C. ambrosioides L. can be an effective and sustainable alternative for the prevention and treatment of diseases caused by various infectious agents.

Fotossensibilização primária em bovinos leiteiros causada por Froelichia humboldtiana / Primary photosensitization in dairy cattle caused by Froelichia humboldtiana

O presente trabalho foi conduzido com o objetivo de relatar um surto de fotossensibilização causado por Froelichia humboldtiana em bovinos leiteiros no Estado do Rio Grande do Norte, Brasil. Foram examinados animais de uma propriedade rural que apresentavam sintomatologia compatível com fotodermatite. Procedeu-se a coleta de amostras de sangue periférico de cinco bovinos para análise das atividades das enzimas hepáticas gamaglutamiltransferase e aspartatoaminotransferase, além da concentração de bilirrubina total, direta e indireta. Das áreas de pele com lesões de dois animais foram realizadas biópsias. Constatou-se que 15 animais de um rebanho composto por 40 animais apresentaram fotossemsibilização. Os animais tinham histórico de apresentar lesões de fotodermatite aproximadamente 10 dias após pastarem em áreas invadidas por F. humboldtiana. Ao exame clínico dos bovinos leiteiros notou-se que inicialmente apresentavam prurido e hiperemia nas áreas de pele despigmentadas do dorso e úbere, também havia alterações do comportamento. Posteriormente, as áreas hiperêmicas se apresentavam com edema que evoluíam para dermatite ulcerativa, necrotizante e exudativa, com perda de extensas áreas da epiderme. As úlceras eram mais graves nos quatro bovinos que apresentavam automutilação por lambedura. Esses quatro animais foram retirados do pasto e abrigados em local sombreado. Uma semana após, o prurido regrediu e as fissuras da pele passaram a cicatrizar. Porém, as lesões reapareceram logo após os bovinos serem reintroduzidas no pasto infestado por F. humbolditiana. Percebeu-se ainda queda na produção leiteira (redução de 50-60%) das vacas após a instalação de fotodermatite. Porém, os bezerros que ainda eram lactantes e ingeriam o leite nas vacas acometidas por fotossensibilização, não apresentaram sinais de fotodermatite. A histopatologia de biópsias de pele revelou inflamação na derme superficial constituída por mastócitos, linfócitos, e alguns plasmócitos. Na epiderme haviam extensas úlceras, recobertas por crostas, associada a infiltrado neutrofílico. As atividades séricas de AST, GGT e as concentrações de bilirrubina estavam dentro dos valores de referência normais para a espécie bovina. O diagnóstico de fotossensibilização primária associada à ingestão de F. humboldtiana foi baseado na epidemiologia, sinais clínicos, bioquímica sérica, biópsia de pele e reocorrência das lesões após os animais serem reintroduzidos no pasto invadido pela planta. Conclui-se que a F. humboldtiana é uma importante causa de fotossensibilização primária em bovinos leiteiros no semiárido brasileiro e que sua toxina provavelmente não é excretada pelo leite bovino.(AU)

The present study was conducted with the objective to report an outbreak of photosensitization caused by Froelichia humboldtiana in dairy cattle in the State of Rio Grande do Norte, Brazil. Animals from a rural property with symptoms compatible with photodermatitis were examined. Peripheral blood samples from five cattle were collected for the analysis of the activities of hepatic enzymes gammaglutamyltransferase and aspartate aminotransferase, in addition were also analysed the concentration of total, direct and indirect bilirubin. From the areas of skin with lesions of two animals, biopsies were performed. It was verified that 15 animals from a herd composed by 40 animals presented photosensitization. The animals had a history of photodermatitis lesions approximately 10 days after grazing in areas invaded by F. humboldtiana. Clinical examination of dairy cattle showed that they initially had pruritus and hyperemia in the depigmented areas of the dorsum and udder, and there were also behavioral changes. Subsequently, the hyperemic areas presented edema that evolved to ulcerative, necrotizing and exudative dermatitis, with loss of extensive areas of the epidermis. The ulcers were more severe in four bovines that had self-mutilation by licking. These four animals were removed from the pasture and sheltered in a shady location. A week later, the pruritus regressed and the fissures of the skin began to heal. However, the lesions reappeared after the cattle were reintroduced in the grass infested by F. humbolditiana. There was also a decrease in milk production (reduction of 50-60%) of cows after the installation of photodermatitis. However, calves that were still lactating and ingested the milk in photosensitized cows, showed no signs of photodermatitis. Histopathology of skin biopsies revealed inflammation in the superficial dermis consisting of mast cells, lymphocytes, and some plasma cells. In the epidermis there were extensive ulcers, covered by crusts, associated with neutrophilic infiltrate. Serum activities of AST, GGT and bilirubin concentrations were within normal reference values for the bovine species. The diagnosis of primary photosensitization associated with F. humboldtiana ingestion was based on epidemiology, clinical signs, serum biochemistry, skin biopsy and lesion reoccurrence after the animals were reintroduced in the pasture invaded by the plant. It is concluded that F. humboldtiana is an important cause of primary photosensitization in dairy cattle in the Brazilian semi-arid region and that its toxin is probably not excreted by bovine milk.(AU)

Chemical constituents from ethyl acetate extracts of domestic Pfaffia glomerata / 中草药

Objective: To study the chemical constituents of ethyl acetate extract from domestic Pfaffia glomerata. Methods: The compounds were isolated and purified by silica gel, reversed-phase, Sephadex LH-20 column chromatography, and their structures were elucidated by spectroscopic analysis as well as chemical methods. Results: Twenty compounds were identified as 1-undecanol (1), oleic acid (2), β-sitosterol (3), 2-hydroxy-4,6-dimethoxyacetophenone (4), oleanolic acid (5), pfaffianol A (6), benzene-1,4-diol (7), vanillic acid (8), iresinone (9), ethylcaffeate (10), oleanoicaeid-28-O-β-D-glucopyranosyl ester (11), allantoin (12), ajugasterone-C (13), β-ecdysterone (14), iresinoside (15), adenine (16), oleanolic acid 3-O-β-D-glucuronopyranoside (17), ficusoside B (18), pfaffiaglycoside B (19), and β-D-Glucopyranosyl-3-(O-β-D-glucopyranosyloxy)-oleanolate (20). Conclusion: Compounds 7-10, 15, 17, 18, and 20 are isolated from this plant for the first time.

Triterpenoid saponins from the roots of Cyathula officinalis and their inhibitory effects on nitric oxide production / 中国天然药物

The present study was designed to investigate the chemical constituents of the roots of Cyathula officinalis. Compounds were isolated by silica gel, Sephadex LH-20, ODS column chromatography, and preparative HPLC. Their structures were determined on the basis of 1D and 2D NMR techniques, mass spectrometry, and chemical methods. One new oleanane-type triterpenoid saponin, 28-O-[α-L-rhamnopyranosyl-(1→3)-β-D-glucuronopyranosyl-(1→3)-β-D-glucopyranosyl] hederagenin (1), was isolated from the roots of Cyathula officinalis. The anti-inflammatory activities of the isolates were evaluated for their inhibitory effects against LPS-induced nitric oxide (NO) production in RAW 264.7 macrophages cells. Compounds 2, 4, and 6 exhibited moderate anti-inflammatory activities.

Anatomical and histochemical analysis of Dysphania ambrosioides supported by light and electron microscopy

ABSTRACT Dysphania ambrosioides (L.) Mosyakin & Clemants (syn: Chenopodium ambrosoides L.), Amaranthaceae, popularly known as “mastruz”, is an herb widely used in Brazil as anthelmintic. To contribute to the knowledge about medicinal plants, a microscopic analysis was accomplished to describe the main anatomical characters of root, stem, petiole and leaf blade of D. ambrosioides and histochemical tests were performed on the leaf blade. Cross-sections were obtained, by hand, for microscopic analysis of root, stem, petiole and leaf blade; to the leaf blade were still made paradermal sections, scanning electron microscopy analysis, maceration and histochemical tests. The main characters useful in the identification of the plant were: anomalous secondary thickening in the root and stem; presence of idioblasts containing crystal sand in the root, stem, petiole and leaf blade; in these there are also idioblasts with druses; presence of non-glandular and glandular trichomes in the stem, petiole and leaf blade; stomata on the stem, petiole and leaf blade, identified in these as anomocytic and anisocytic; dorsiventral mesophyll and collateral vascular bundles. Maceration revealed that the vessel elements are helical type. Through the histochemical tests, it was evidenced the presence of lipophilic substances, essential oils, oleoresins, phenolic compounds, starch, lignin and calcium oxalate crystals. This work provides support to the quality control of the species.

Troponina C na detecção imuno-histoquímica de alterações regressivas precoces no miocárdio de ovinos naturalmente intoxicados por Amaranthus spinosus (Amaranthaceae) / Troponin C in the immunohistochemistry detection of early regressive changes in sheep myocardium naturally poisoned by Amaranthus spinosus (Amaranthaceae)

Amaranthus spp. são plantas nefrotóxicas popularmente conhecidas como "caruru". Em casos de intoxicação por estas plantas, a principal alteração histopatológica está presente no rim, sob forma de nefrose tubular tóxica, porém em alguns casos pode haver alterações cardíacas. Alterações no eletrocardiograma, compatíveis com quadros de hipercalemia, foram descritas em suínos intoxicados por Amaranthus retroflexus e lesões como degeneração e necrose de miócitos cardíacos descritas em suínos intoxicados por A. caudatus e ovinos intoxicados por A. spinosus. Há dúvidas com relação às alterações cardíacas, que, na maioria dos casos, são incipientes, o que pode levar a erros de interpretação. Para a realização do trabalho foram utilizados blocos parafinados oriundos de um surto natural de intoxicação por A. spinosus no sudeste do Brasil. Esse estudo teve como objetivo detectar a presença de alterações regressivas incipientes no miocárdio de ovinos intoxicados por A. spinosus, através da utilização imuno-histoquímica do anticorpo anti-troponina C. Foram utilizados fragmentos de coração de 8 ovinos adultos e 2 fetos, intoxicados naturalmente por A. spinosus. Estes fragmentos foram submetidos à técnica de imuno-histoquímica com a utilização do anticorpo anti-troponina C. Pela avaliação imuno-histoquímica do coração dos oito ovinos adultos observaram-se diversos grupos de miócitos com diminuição significativa ou ausência de imunorreatividade para o anticorpo anti-troponina C; essas áreas correspondiam, em grande parte, aos mesmos grupos de miócitos que apresentavam, pela coloração de Hematoxilina e Eosina (H.E.) alterações que variavam de leve tumefação celular a aumento da eosinofilia, perda de estriação, lise celular e cariólise, ou mais raramente, acompanhadas de infiltrado inflamatório...

Amaranthus spp. are nephrotoxic plants popularly known as "pigweed". In cases of poisoning by these plants, the main histopathological alteration is found in the kidneys as toxic tubular nephrosis; however, in some cases, there may be cardiac changes. ECG changes associated with hyperkalemia have been described in pigs poisoned by Amaranthus retroflexus. Degeneration and necrosis of myocytes have been described in pigs poisoned by A. caudatus and sheep poisoned by A. spinosus. There are doubts regarding cardiac changes, since in most cases they are incipient and don't exhibit inflammatory reaction, which can lead to misinterpretation. For this study, paraffin blocks with tissues from a poisoning outbreak by A. spinosus in southeastern Brazil were used. The objective of the study was to detect the presence of incipient regressive changes in the myocardium of sheep poisoned by A. spinosus using anti-troponin C antibody-based immunohistochemistry. Fragments of hearts from 8 adult sheep and 2 fetuses naturally poisoned by A. spinosus were used. In the immunohistochemistry evaluation of the 8 hearts from the adult sheep there were several groups of myocytes with significant decrease or absence of immunoreactivity for anti-troponin C antibody. In most cases, these same areas on Hematoxylin and Eosin (HE) staining exhibited changes that varied from mild cellular tumefaction to increased eosinophilia, as well as loss of striation, cell lysis and karyolysis, sometimes accompanied by inflammatory infiltrate...

Preliminary pharmacognostic screening of Achyranthes coynei stem.

Achyranthes coynei is a rare, endemic perennial shrub reported from Karnataka and Maharashtra states of India. The plant is used to treat various disorders by folk healers and was proven to have antimicrobial and antioxidant properties. The present study was undertaken to evaluate microscopic and macroscopic characters of A. coynei stem, along with its physicochemical parameters. ProgRes® CapturePro and Microsoft Excel were used for statistical analysis. Perennial, shrubby nature and woody stem were the distinguishing morphological characters observed. Transverse section (TS) illustrated quadrangular outline of the stem and showed the presence of two types of trichomes on the thick‑walled epidermis. TS also showed number of rosette calcium oxalates crystals; prismatic and microsphenoid crystals; conjoint, collateral open secondary vascular bundles; and two amphixylic medullary bundles in the pith. Ash and extractive values, micro and macro elements and nutritive factors were estimated in the present study. The presence of alkaloids, saponins and triterpenoids were observed in preliminary phytochemical screening. High‑performance thin layer chromatographic analysis yielded different bands and also indicated the presence of oleanolic acid. The studied parameters for A. coynei stem will be useful for identification and authentication of the plant material.

In vitro Callus Induction and Plant Regeneration of Celosia argentea- An Important Medicinal Plant

Celosia argentea (Var.) cristata (Amaranthaceae) is a widely cultivated ornamental plant, which has antibacterial, astringent, haemostatic, hypertensive, ophthalmic, and parasitic significance. This study describes a protocol for in vitro callus induction and plant regeneration from leaf and stem explants of C. argentea using Murashige and Skoog (MS) medium. Callus culture was initiated and established from seedling, leaf, and stem explants. Explants were cultured on MS medium supplemented with auxin alone (0.5 mg/L Naphthaleneacetic acid (NAA), 2, 4-Dicholorophenoxyacetic acid (2, 4-D). Green and red compact callus (98%) were induced using MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L Benzyladenine (BA). Two different concentrations (1.0 mg/L BA + 0.5 mg/L NAA and 1.0 mg/L BA + 1.0 mg/L NAA) successfully induced plant regeneration with multiple shoots (1.5 and 0.9 shoots per explants, respectively). Successful shoots were transferred to rooting medium supplemented with 1.0 mg/L Indole-3-acetic acid (IAA) (80%) at 35th day. Acclimatization was done, which resulted in 90% of the plantlets surviving in garden soil. This protocol could be used to micropropagate C. argentea for conservation, commercial natural product production. The high frequency of callus indicated potential of C. argentea for secondary metabolite production (celosin) in pharmaceutical industry.

Phytochemistry and hepatoprotective activity of aqueous extract of Amaranthus tricolor Linn roots.

Background: The genus Amaranthus has potential activity as a hepatoprotective agent. Objective: The present pharmacological investigation focuses on evaluation of the effi cacy of aqueous extract of roots of Amaranthus tricolor Linn. for their protection against paracetamol (PCM) overdose induced hepatotoxicity. Materials and Methods: The aqueous extract of roots of A. tricolor Linn. was prepared and phytochemical screening was done. The biochemical investigation viz. serum glutamic oxaloacetate transaminase (SGOT), serum glutamic pyruvate transaminase (SGPT), alkaline phosphatase (ALP) and total Bilirubin (TB) was done against PCM-induced hepatotoxicity in wistar albino rats. The histopathological studies of liver were also done. Results: The phytochemical screening of the aqueous extract showed the presence of alkaloids, carbohydrates, fl avanoids, amino acids, proteins, fi xed oil, saponins and tannins, and phenolic compounds. Pretreatment with the aqueous extract of root signifi cantly prevented the physical, biochemical, histological, and functional changes induced by paracetamol in the liver. The extract showed signifi cant hepatoprotective effects as evidenced by decreased serum enzyme activities like SGPT, SGOT, ALP, and TB, which was supported by histopathological studies of liver. The aqueous extract showed signifi cant hepatoprotective activity comparable with standard drug silymarin as well as hepatotoxin drug PCM. Conclusion: From these results, it is concluded that the A. tricolor has potential effectiveness in treating liver damage in a dose dependent manner.

Intoxicação espontânea por Amaranthus hybridus (Amaranthaceae) em bovinos no sul do Rio Grande do Sul / Spontaneous poisoning by Amaranthus hybridus in cattle in southern Rio Grande do Sul

Descreve-se um surto de intoxicação por Amaranthus hybridus ocorrido em bovinos em fevereiro/março de 2013 na região sul do Rio Grande do Sul. A morbidade foi de 48,33%, a mortalidade de 41,66% e a letalidade 86,20%. O diagnóstico da intoxicação por A. hybridus foi baseado nos dados epidemiológicos, sinais clínicos, achados de necropsia e lesões histológicas características da intoxicação por plantas nefrotóxicas e pela presença da planta em grande quantidade na área onde estavam os bovinos. A invasão da pastagem de Brachiaria sp. pela planta alerta para a possibilidade da ocorrência da intoxicação não somente em áreas de resteva com carência de forragem, uma vez que no surto relatado neste trabalho havia abundante disponibilidade de forragem nas áreas onde os bovinos se intoxicaram. No presente caso provavelmente as boas condições climáticas e o solo fertilizado para a plantação da pastagem favoreceram o crescimento da invasora.

An outbreak of Amaranthus hybridus poisoning in cattle was observed in February/March 2013 in southern Rio Grande do Sul, Brazil. The morbidity rate observed was 48.33%, the mortality rate was 41.66% and lethality rate was 86.20%. The diagnosis of A. hybridus poisoning was made based on the epidemiological data, clinical signs, macroscopic and histological lesions characteristic of intoxication by nephrotoxic plants and by the presence of large amounts of the plant in the area where cattle were placed. Invasion of Brachiaria sp. pasture by A. hybridus alerts for the possibility of the occurrence of poisoning not only in stubble areas with lack of forage. In the case reported here there was abundant forage availability in the area where the outbreak occurred. Favorable climatic conditions and fertile soil for planting pasture, probably favored the growth of this invasive species.

Isolation of alpha-spinasterol from Amaranthus spinosus stems / Aislación de alfa-spinasterol de ramas de Amaranthus spinosus

alpha spinasterol was identified in stems of Amaranthus spinosus. The structure was obtained by spectroscopic methods and comparison with literature data.

alfa spinasterol fue identificado en ramas de Amaranthus spinosus. La estructura fue obtenida por metodos espectroscópicos y por comparación con datos de literature.

Biological screening of Amaranthus retroflexus L. (Amaranthaceae) / Abordagem biológica de Amaranthus retroflexus L. (Amaranthaceae)

Bioassays are required for the determination of the total toxicity of Amaranthus retroflexus L. (Amaranthaceae) or "redroot pigweed". Therefore, the plant extract has been tested for bioactivity in Artemia salina and cytotoxicity against bovine kidney cells. The LD50 values for Artemia salina were measured at 1700 ppm. The bovine kidney cells were exposed to various concentrations of the plant extracts (100 ppm-0.1 ppm). After treating with 100 and 0.1 ppm for 24 h, the cells viability were reduced by about 49 percent and 35 percent respectively in MTT viability assay. The study confirmed that Amaranthus retroflexus has a cytotoxic effect and more specific to renal cells.

Ensaios biológicos foram realizados para determinação da toxidade de Amaranthus retroflexus L. (Amaranthaceae) conhecido popularmente como "redroot pigweed". Extratos desta espécie foram testados para avaliar sua bioatividade em Artemia salina e citoxidade em células bovinas de rim. Os valores de DL50 para Artemia salina foram medidos a 1700 ppm. As células de rim bovinas foram expostas a várias concentrações dos extratos de plantas (100-0,1 ppm). Após tratamento com 100 e 0,1 ppm por 24 h, a viabilidade celular foi reduzida a cerca de 49 por cento e 35 por cento, respectivamente, no ensaio de MTT. O estudo confirma que Amaranthus retroflexus apresenta um efeito citotóxico e, mais especificamente, para células renais.

pH do meio de cultivo e crescimento de plântulas de ginseng brasileiro cultivadas in vitro / Culture medium pH and growth of brazilian ginseng in vitro cultured plantlets

O presente trabalho objetivou avaliar o efeito do pH do meio de cultivo sobre alguns parâmetros de crescimento da Pfaffia glomerata (Spreng.) Pedersen cultivada in vitro, bem como checar se o crescimento dos explantes altera o pH do meio ao longo do período de cultivo. Foram testados quatro tratamentos constituídos de distintos valores de pH (3,7; 5,0; 6,0 e 7,5) do meio de cultivo. O pH do meio de cultivo foi ajustado antes da inclusão do agar (6g L-1 - Merck) e da autoclavagem. Como fonte de explantes foram utilizadas segmentos nodais de plantas previamente estabelecidas in vitro em meio MS. Dos nove aos 15 dias após a inoculação (DAI) dos segmentos nodais, verificou-se maior número de raízes em pH 6,0 e o menor no pH 7,5. Aos 35 DAI, o comprimento da maior brotação e o número total de segmentos nodais por planta foram maiores em torno de pH 6,0. Aos 35 DAI, observou-se menor crescimento em biomassa de raízes em pH 3,7. Já a parte aérea apresentou menor biomassa em pH 7,5. Aos 35 DAI, a produção de matéria fresca e seca total da plântula foi maior em pH próximo a 6,0. Concluiu-se que valores de pH do meio de cultivo próximos a 6,0, ajustados antes da autoclavagem, são ideais para o crescimento da P. glomerata cultivada in vitro. Também se verificou que o crescimento da plântula modificou significativamente o pH do meio de cultivo.

The present research aimed to evaluate the effect of culture medium pH on some growth parameter of Pfaffia glomerata (Spreng.) Pedersen in vitro cultured plantlets, as well as to check whether the explant´s growth alters the culture medium pH. Four treatments consisted of different values (3.7; 5.0; 6.0 and 7.5) of culture medium pH were tested. The culture medium pH was adjusted prior to the addition of agar (6g L-1 - Merck) and autoclaving. Nodal segments from asseptic plants grown in MS medium were used as explants. From 9 to 15 days after inoculation (DAI) of nodal segments, the higher number of roots was obtained at pH 6.0, and the lower at pH 7.5. At 35 DAI, both length of the higher sprout and total number of nodal segments per plantlet were greater at about pH 6.0. At 35 DAI, roots biomass was lower at pH 3.7. On the other hand, shoots biomass was lower at pH 7.5. Fresh and dry matter of the whole plantlet was greater at pH around 6.0. In conclusion, values of culture medium pH near to 6.0, adjusted prior to autoclaving, are ideal for the growth of P. glomerata in vitro cultured plantlets. Moreover, the in vitro growth of plantlet modified significantly the culture medium pH.

Evaluation of immunomodulatory and anti-inflammatory effects and phytochemical screening of Alternanthera tenella Colla (Amaranthaceae) aqueous extracts

Alternanthera tenella Colla extracts are used in Brazilian traditional folk medicine to treat a variety of infectious diseases as well as inflammation and fever. In this work, the immunomodulatory, anti-inflammatory and potential toxic effects of cold (CAE) and hot (HAE) aqueous extracts of A. tenella were investigated in vivo. In addition, we analyzed the phytochemical properties of both extracts. BALB/c mice were immunized in vivo with sheep red blood cells and concomitantly inoculated intraperitoneally (i.p.) with each extract (50, 100 or 200 mg/kg). Specific antibody-producing cells were enumerated using plaque-forming cell assays (PFC) and anti-SRBC IgG and IgM serum levels were measured via enzyme-linked immunosorbent assay. Body and lymphoid organ weights were determined after treatments in order to evaluate toxic effects. Carrageenan-induced paw edema was employed to investigate anti-inflammatory activity in mice inoculated i.p. with CAE or HAE (200 or 400 mg/kg). Phytochemical screening was performed using spectrometric and chromatographic approaches and revealed that CAE possessed higher tannin and flavonoid levels than HAE. PFC numbers were increased after treatment with CAE (100 mg/kg) four days after immunization, as were the serum antibody titers after four and seven days, suggesting immunostimulatory activity through modulation of B lymphocyte functions. Body and organ weights did not show major changes, suggesting that extracts administered to mice did not induce significant toxicity. Both extracts had significant anti-inflammatory activity in the paw edema assay. These results suggested that aqueous extracts from A. tenella contained several chemical compounds that possess positive and/or negative modulator effects on the immune system, which appeared to correlate with tannin and flavonoid levels in those extracts. In summary, these studies provide important insight into the biological activities of A. tenella.

Caracterização das exigências nutricionais de mudas de Pfaffia glomerata em Argissolo Vermelho distrófico arênico pela técnica do nutriente faltante / Characterization of the nutritional demands of young plants of Pfaffia glomerata in a Paleudalf soil by means of the missing nutrient technique

Este trabalho teve por objetivo caracterizar as exigências nutricionais de mudas de ginseng brasileiro (Pfaffia glomerata (Spreng.) Pedersen) em Argissolo Vermelho distrófico arênico pela técnica do nutriente faltante. Plantas de P. glomerata produzidas in vitro e aclimatizadas ex vitro foram usadas como material inicial para o experimento em casa de vegetação. Aos 10 dias após o transplante para o solo, através de soluções nutritivas, foram realizadas as adubações correspondentes a nove tratamentos: controle (sem adubação), adubação completa (com macro e micronutrientes) e as omissões individuais de N, P, K, Ca, Mg, S e dos micronutrientes (Zn, B, Cu, Fe, Mn e Mo). Aos 60 dias de cultivo, avaliaram-se os parâmetros de crescimento das plantas. O número de folhas por planta foi o parâmetro de crescimento mais afetado pela ausência dos nutrientes N, K, S e P na adubação. O efeito da omissão dos nutrientes na adubação foi diferenciado quanto à partição de biomassa entre os órgãos da planta. A biomassa acumulada pela parte aérea obedeceu à seguinte ordem decrescente, em relação à omissão do nutriente: Mg aproxidamente micronutrientes > Ca aproximadamente adubação completa > P > S > K > N < aproximadamente controle. Já a biomassa radicular obedeceu à seguinte ordem decrescente: Mg aproximadamente adubação completa aproximadamente ≥P ≥ micronutrientes ≥ S ≥ controle. Plantas jovens de P. glomerata cultivadas em Argissolo Vermelho distrófico arênico apresentaram grande redução no crescimento pela ausência de N, K, S e P na adubação. As omissões de Mg e dos micronutrientes na adubação não foram limitantes para o crescimento das plantas jovens.

This study aimed to characterize the nutritional demands of the young plants of Brazilian ginseng (Pfaffia glomerata (Spreng.) Pedersen) in a Paleudalf soil by means of the missing nutrient technique. Plantlets of P. glomerata produced in vitro and acclimatized ex vitro were used as start matter for the glasshouse experiment. Ten days after plant transfer to soil, fertilizations (by means of nutritive solutions) were done corresponding to nine treatments: control (without fertilization), complete fertilization (with macro and micronutrients), and with the omissions of N, P, K, Ca, Mg, S and micronutrients (Zn, B, Cu, Fe, Mn and Mo). At 60 days of cultivation, several growth parameters were evaluated. The number of leaves per plant was the growth parameter most affected by omission of N, K, S and P in the fertilization. The effect of nutrient omission in the fertilization on biomass partitioning between plant organs were differentiated. Accumulated shoot biomass obeied the following decreasing order, in relation with the nutrient omission: Mg approximatelly micronutrients > Ca approximatelly > P > S > K > N approximatelly control. While the root biomass obeied the following decreasing order: Mg approximatelly complete fertilization ≥ P ≥ micronutrients ≥ S ≥ Ca > N > K < approximatelly control. Young plants of P. glomerata grow in a Paleudalf soil showed greater reduction in growth by omission of N, K, S and P in the fertilization. The omission of Mg and micronutrients in the fertilization were not limiting to the growth of young plants.

Hypnotic effect of ecdysterone isolated from Pfaffia glomerata (Spreng.) Pedersen / Efeito hipinótico de ecdisterona isolada de Pfaffia glomerata (Spreng.) Pedersen

Neste trabalho foi avaliado, em roedores, o efeito depressor das frações clorofórmio (CHCl3), acetato de etila (EtOAc) e n-butanol, obtidas das partes subterrâneas de Pfaffia glomerata, empregando-se o teste de tempo de sono barbitúrico como referência. Somente a fração lipofílica (CHCl3:EtOAc, 1:1, m/m) (i.p. 500 mg/kg; v.o. 1000 mg/kg) potenciou o tempo de sono induzido por pentobarbital. A ecdisterona foi isolada e identificada como constituinte majoritário (1,4 por cento m/m) desta fração, através de cromatografia líquida de alta eficiência e métodos espectroscópicos, respectivamente. Este composto potenciou o tempo de sono barbitúrico (100 mg/kg, i.p.; 400 mg/kg, v.o), sem causar hipotermia. Nestas mesmas doses, a ecdisterona não alterou a performance dos animais no rota-rod, esquiva inibitória e labirinto em cruz-elevado, além de não alterar o padrão de convulsões induzidas por pentilenotetrazol. Este perfil indica que esta substância, nestas doses, não apresenta perfil ansiolítico ou neurotóxico. Estes resultados indicam que a ecdisterona é o componente responsável pela ação hipnótica apresentada pela fração lipofílica obtida das partes subterrâneas de P. glomerata.

In this study the depressant effect of fractions from P. glomerata was initially evaluated using the mice barbiturate sleeping time test as reference. The fractions tested were the CHCl3, the EtOAc, the n-BuOH and the aqueous fraction obtained from P. glomerata subterraneous parts. Only the pretreatment with the lipophilic fraction (CHCl3: EtOAc, 1:1, w/w) increased the barbiturate sleeping time (i.p 500 mg/kg; v.o. 1000 mg/kg). Ecdysterone, the main substance isolated from this lipophilic fraction, was identified by spectroscopic methods and its content in the ethanol extract was determined as 1.4 percent (w/w) by HPLC. In order to investigate the hypothesis of ecdysterone displaying a depressant effect on nervous central system, an evaluation toward the hypnotic-sedative and anxiolytic effects of this drug was carried out. Ecdysterone 100 mg/kg, i.p, increased the barbiturate sleeping time without provoking hypothermia; when administered by oral route its minimal effective dose was 400 mg/kg. On the other hand, ecdysterone (100 mg/kg, i.p; 400 mg/kg, p.o) did not impair motor coordination and was ineffective on pentylenetetrazole-induced convulsion, elevated plus-maze and step-down inhibitory avoidance tests, indicating that at these doses the drug does not present an anxiolytic profile and does not cause manifest neurotoxic effects as well. In conclusion, the lipophilic fraction from P. glomerata presents a hypnotic effect being ecdysterone one of the compounds responsible for this CNS activity.