Exopolysaccharides from Klebsiella oxytoca: anti-inflammatory activity

Abstract Exopolysaccharides (EPS) produced by Klebsiella oxytoca are of environmental, pharmaceutical, and medicinal interest. However, studies about the anti-inflammatory activity of EPS produced by this microorganism still remain limited. The aim of this study was to produce, characterize, and evaluate the anti-inflammatory activity of EPS from K. oxytoca in a pleurisy model. Colorimetric analysis revealed that precipitated crude exopolysaccharides (KEPSC) and deproteinated exopolysaccharides (KEPS) present high levels of total carbohydrates (65.57% and 62.82%, respectively). Analyses of uronic acid (7.90% in KEPSC and 6.21% in KEPS) and pyruvic acid (3.01% in KEPSC and 1.68% in KEPS) confirm that the EPS are acidic. Gas chromatography-mass spectrometry analyses demonstrated that the EPS consisted of rhamnose (29.83%), glucose (11.21%), galactose (52.45%), and mannose (6.50%). The treatment of an experimental pleurisy model in rats through subcutaneous administration of 50, 100, 200, and 400 mg/kg of KEPS decreased both the volume of inflammatory exudate and the number of leukocytes recruited to the pleural cavity. The present data showed that EPS production by K. oxytoca using the method described is easy to perform and results in a good yield. In addition, we show that KEPS exhibit anti-inflammatory activity when administered subcutaneously in rats.

Limonoid detection and profile in callus culture of sweet orange

Plant tissue culture has emerged as an important tool to produce bioactive compounds from various plant species, including the sustainable production of limonoids that are receiving considerable attention due to the benefits associated with human health such as anticancer activities. The purpose of the present study was to evaluate the capacity of limonoids aglycone production from callus culture from sweet orange cv. Pera (Citrus sinensis) seeds and identify the compounds produced in this cell line. Callus induction occurred in Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic (2,4-D), malt extract, agar and coconut water. For the analysis and identification of the limonoids, CG-MS-EI ion-positive mode and UPLC-QTOF-ESI were used operating in positive and negative mode. An intense peak corresponding to limonin appeared in the callus extracts at a retention time of 58.1 min. in CG-MS-EI and four major limonoids aglycone by positive ion mode UPLC-QTOF-ESI: limonin, nomilin, deacetylnomilin, and nomilinic acid. The culture medium was efficient at the bioproduction of limonoids aglycone in callus cultures of C. sinensis seeds. Therefore, data obtained from UPLC-QTOF-ESI proved its importance at identifying new compounds that benefit human health, and may assist future work in the identification of known or new limonoids in Citrus species and related genera.

Evaluation of limonoid production in suspension cell culture of Citrus sinensis

ABSTRACTThe use of cell and plant tissue culture techniques to produce economically important active metabolites has been growing. Among these substances are total limonoid aglycones, which are produced by "pera" orange (Citrus sinensis (L.) Osbeck, Rutaceae) and have received considerable attention because of their anticancer actions. The main objective of the present study was to analyze and compare the levels of limonoid aglycones in seeds, callus cultures (originating from seeds), callus cultures (originating from hypocotyls), cell suspensions from hypocotyls cells, and cell suspensions from cotyledons. The cell cultures or C. sinensis were obtained by inoculating two strains of callus in MS medium supplemented with 2.0 µM 2,4-dichlorophenoxyacetic acid, 7.0 µM benzyl aminopurine, and 3% (w/v) sucrose in the dark. The highest concentrations of limonoid aglycone that were obtained were observed in cotyledon cell lines (240 mg/100 g dry weight) that were produced on day 21 of culture and hypocotyl cell lines on day 7 (210 mg/100 g dry weight). Explants of different origins under the same culture conditions had different limonoid aglycone content. The present results may suggest strategies for enhancing the productivity of biologically important limonoid aglycones and investigating the complex pathways of these secondary metabolites in plant tissue cultures.

Produção de metabólitos secundários em cultura de células e tecidos de plantas: o exemplo dos gêneros Tabernaemontana e Aspidosperma: [revisão] / Production of plant secondary metabolites in plant cell and tissue culture: the example of Tabernaemontana and Aspidosperma genera: [revision]

Os estudos dos metabólitos secundários de plantas se desenvolveram aceleradamente nos últimos 50 anos. Estes compostos são conhecidos por desempenharem um papel importante na adaptação das plantas aos seus ambientes e também representam uma fonte importante de substâncias farmacologicamente ativas. As técnicas de cultura de células de plantas iniciaramse na década de 1960 como uma possível ferramenta para estudar e produzir os metabólitos secundários de plantas. O uso de cultura de células de planta para a produção de substâncias de interesse contribuiu grandemente para avanços em diversas áreas da fisiologia e bioquímica vegetal. Diferentes estratégias, usando sistemas de cultura in vitro, foram estudadas com o objetivo de aumentar a produção de metabólitos secundários. As plantas dos gêneros Aspidosperma e Tabernaemontana são importantes fontes de alcalóides indólicos biologicamente ativos, sendo que no Brasil existe um número considerável de espécies destes gêneros. As culturas de células de Aspidosperma e Tabernaemontana foram iniciadas há pelo menos 16 anos, as quais produzem um grande número de alcalóides, o que estimulou o desenvolvimento de diversas técnicas para sua produção, extração e identificação.

Studies on plant secondary metabolites have been increasing over the last 50 years. These compounds are known to play a major role in the adaptation of plants to their environment and an important source of active pharmaceuticals. Plant cell culture technologies were introduced at the end of the 1960s as a possible tool for both studying and producing plant secondary metabolites. Different strategies, using in vitro systems, have been extensively studied with the objective of improving the production of secondary plant compounds. The Aspidosperma and Tabernaemontana genera are an important source of biologically active alkaloids and in Brazil there is a considerable number of species of these genera. About 16 years ago cell cultures of Tabernemontana and Aspidosperma were initiated. These cell cultures did produce a number of alkaloids of pharmaceutical interest that stimulated the development of several techniques to production, extraction and identification.

Comparação de diferentes metodologias para obtenção de cultura de calos de Stevia rebaudiana (Bertoni) Bertoni / Comparison of different methodologies for obtaining callus cultures from Stevia rebaudiana (Bertoni) Bertoni

As folhas de Stevia rebaudiana (Bertoni) Bertoni (Asteraceae) contêm glicosídeos diterpenóides (GDS), que são cerca de 300 vezes mais doce que a sacarose a 4%. O objetivo deste estudo foi avaliar a formação de calos, a partir de folhas obtidas in vivo e in vitro de S. rebaudiana em dois meios já descritos na literatura: Murashige e Skoog (MS), suplementado com 3 mg L-1 de ácido 2,4-diclorofenóxiacético (2,4-D), e o MS suplementado com 1 mg L-1 de ácido naftalenoacético (ANA) e 0,5 mg L-1 de 6-benzilaminopurina 6-BAP) e um desenvolvido em nosso laboratório o Woody Plant Medium (WPM), suplementado com 6 mg L-1 de ANA e 4 mg L-1 de cinetina (CIN). Os explantes obtidos in vitro iniciaram a formação de calos um pouco mais rapidamente que os das folhas de plantas advindas da natureza. A utilização dos nutrientes do meio WPM, associada a uma combinação de fitorreguladores adequada, proporcionou velocidade de indução e multiplicação de calos bem maiores que as apresentadas nos meios que empregaram os nutrientes do MS. Novos experimentos serão realizados, depois de alcançada a estabilidade genética dos calos, visando avaliar a capacidade destes em biossintetizar os GDS

The leaves from Stevia rebaudiana (Bertoni) Bertoni (Asteraceae) contain diterpenoid glycosides (GDS), which are almost 300 times sweeter than sucrose at 4%. The subject of this study was to evaluate the callus-formation from in vivo and in vitro leaves of Stevia rebaudiana in two already described in literature: Murashige and Skoog (MS) supplemented with 3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D); MS supplemented with 1 mg L-1 of naphthaleneacetic acid (NAA) and 0.5 mg L-1 of 6-benzylaminopurine (6-BAP) and other developed in our laboratory the Woody Plant Medium (WPM) with 6 mg L-1 of NAA and 4 mg L-1 of Kinetin (KIN). The explants obtained in vitro initiated callus formation faster than leaves from natural plants. The utilization of WPM nutrients, associated with an adequate combination of phytoregulators, provided greater callus induction velocity and multiplication than the media that using MS nutrients. New experiments will be conducted after reaching genetic stability of the calluses, seeking to evaluate the capacity of these calluses to biosynthesize GDS

Multiplicação in vitro de Aspidosperma ramiflorum Muell. Arg. (Apocynaceae) / In vitro multiplication of Aspidosperma ramiflorum Muell. Arg. (Apocynaceae)

O presente estudo relata um método simples e promissor para multiplicação in vitro de Aspidosperma ramiflorum, uma espécie encontrada no sudeste do Brasil e seriamente ameaçada de extinção, utilizada com propósitos medicinais e como fonte de compostos que podem ser usados para desenvolver novos fármacos sintéticos. O trabalho teve como objetivo o estabelecimento de um protocolo de multiplicação in vitro de Aspidosperma ramiflorum (guatambu), a partir de segmentos apicais de material juvenil originários de plântulas obtidos a partir de sementes. A avaliação da multiplicação in vitro foi realizada em meio de cultura Woody Plant Médium (WPM), suplementado com concentrações variadas de ácido naftalenoacético (ANA) e 6-Benzilaminopurina (6-BAP). A multiplicação de A. ramiflorum foi positivamente influenciada principalmente nas combinações aonde as concentrações de 6-BAP foram relativamente maiores do que as do ANA, nessas concentrações houve a indução de múltiplas brotações.

The present study described a simple and promissory method for in vitro multiplication of Aspidosperma ramiflorum, a species found in the South of Brazil and seriously extinction menaced. The method was used for medicinal proposes and as a source of compounds to develop new synthetic drugs. The objective of this work was to establish an in vitro multiplication protocol of Aspidosperma ramiflorum (guatambu), from apical segments of juvenile material of plantlets obtained from seeds. The in vitro multiplication evaluation was done in WPM medium, supplemented with variable concentrations of Naphthalene acetic acid (NAA) and 6- Benzyl aminopurine (6-BAP). The multiplication of A. ramiflorum was positively influenced mainly in the combinations when 6-BAP concentrations were relatively higher than NAA. In these concentrations multiple shoots were induced.

Avaliação do controle de qualidade realizado nas farmácias de manipulação e homeopáticas de Maringá, Estado do Paraná / Evaluation of quality control in drug-manipulation and homeopathic pharmacies in Maringá, Paraná State

O controle de qualidade realizado nas farmácias de manipulação e homeopáticas é de suma importância para assegurar a qualidade microbiológica e físico-química das matérias-primas e produtos acabados, garantindo eficácia, segurança e credibilidade dos medicamentos dispensados à população. O presente trabalho teve como objetivos realizar uma pesquisa sobre o número de farmácias de manipulação e homeopáticas da cidade de Maringá, Estado do Paraná, que estão realizando o controle de qualidade de acordo com o estabelecido pela Vigilância Sanitária na Resolução de Diretoria Colegiada (RDC33); diagnosticar quais os testes mínimos realizados; observar a adequação das técnicas empregadas e quais as maiores dificuldades encontradas pelas farmácias para implantar o controle de qualidade. Com base nas diretrizes da RDC33 foi formulado um questionário, o qual foi utilizado como suporte para a realização da pesquisa direta em 25 farmácias de manipulação e homeopáticas, que aceitaram participar do estudo, de um total de 30 farmácias da cidade de Maringá. A partir das análises realizadas, concluiu-se que a maioria das farmácias de manipulação e homeopáticas da cidade de Maringá está se adequando às exigências estabelecidas pela Anvisa. Porém, encontram dificuldades como recursos financeiros e humanos para realizarem o controle de qualidade mínimo necess??rio para a manutenção da eficácia, estabilidade e segurança dos medicamentos manipulados e matérias-primas utilizadas.

The quality control in compounding and homeopathic pharmacies is of highest importance to assure the microbiological and physical-chemical quality of drugs and compounding preparations, guaranteeing effectiveness, safety and credibility of the released medicines to the population. The objective of the present work was to verify the number of compounding and homeopathic pharmacies of the city of Maringá, Paraná State, which are doing the quality control in agreement with the determinations of the Sanitary Surveillance in RDC33. It was observed which minimum tests, were realized and if they were adequate as well the largest difficulties found by the drugstores to implant the quality control. Using as guideline the RDC33 a questionnaire was formulated, which was used as support for the accomplishment of the direct research in 25 compounding and homeopathic pharmacies from a total of 30 drugstores of the city of Maringá. Based analyses of the results it was concluded that most of the compounding and homeopathic pharmacies of the city of Maringá are trying to adaptate to the established demands of ANVISA. However they have difficulties such as financial and human resources to do the minimum quality control necessary for the maintenance of the effectiveness, stability and safety of the manipulated medicines and raw materials used.