Estabilidad genética entre clones de berenjena in vitro inducidos por diferentes reguladores de crecimiento de plantas

Many factors may influence the genetic stability of plant in vitro clones, among which the genotype and the regenerative process induced by plant growth regulators. The resulting somaclonal variations may be useful for breeding projects, but may be detrimental to germplasm conservation. The objective of this work was to evaluate the genetic stability of Solanum melongena cv. Florida Market clones, obtained in response to different plant growth regulators. For the production of clones, leaf explants were used from commercial seed germinated plants. The explants were inoculated in Murashige and Skoog medium supplemented with different plant growth regulators at pre­defined concentrations. The DNA was extracted by the CTAB method from leaves of complete plants obtained by somatic embryogenesis induced by naphthaleneacetic acid (NAA) or indirect organogenesis induced by benzylaminopurine (BAP) or thidiazuron (TDZ). For the RAPD, 117 DNA samples were amplified by ten decamer primers and 49 specific bands were selected among the products for the comparative study. A total of 5733 fragments were obtained, with a rate of 5.37% polymorphism. NAA did not generate polymorphism and the BAP was responsible for the highest rate obtained (14.28%). Two RAPD primers were identified as markers for monitoring the genetic stability of eggplant. The polymorphic pattern was observed only in clones originating from indirect organogenesis. These results indicate the usefulness of a monitoring protocol for studies using in vitro cloned eggplant.Muchos factores pueden influir en la estabilidad genética de los clones de plantas in vitro, entre los que se encuentran el genotipo y el proceso regenerativo inducido por los reguladores del crecimiento. Por lo tanto, las variaciones somaclonales resultantes del cultivo pueden ser útiles para proyectos de mejoramiento genético, pero pueden ser perjudiciales para la conservación de germoplasma. El objetivo de este estudio fue evaluar la estabilidad genética de Solanum melongena cv. Florida Market, obtenida en respuesta a diferentes reguladores del crecimiento. Para la producción de clones se utilizaron explantes de hojas provenientes de plantas obtenidas de semillas germinadas. Los explantes se inocularon en medio de cultivo Murashige y Skoog con los diferentes reguladores del crecimiento en concentraciones predefinidas. El ADN se extrajo mediante el método CTAB a partir de plantas completas obtenidas por medio de embriogénesis somática inducida por ácido naftalenoacético (ANA) u organogénesis indirecta inducida por bencilaminopurina (BAP) o tidiazurón (TDZ). Para el RAPD, 117 muestras de ADN se amplificaron mediante diez cebadores y se seleccionaron 49 bandas puntuales entre los productos, para el estudio comparativo. Se obtuvieron un total de 5733 fragmentos, con una tasa de 5.37% de polimorfismo. ANA no generó polimorfismo y BAP fue responsable de la tasa más alta obtenida (14.28%). Se identificaron dos cebadores RAPD como marcadores para monitorear la estabilidad genética de la berenjena. El patrón polimórfico se observó solo en los clones originados en la organogénesis indirecta. Estos resultados indican la utilidad de un protocolo de monitoreo para estudios que usan berenjena clonada in vitro. Palabras clave: Solanum melongena, cultivo in vitro, variación somaclonal, RAPD, polimorfismo de ADN, fitomejoramient

CONSERVAÇÃO IN VITRO DA BIODIVERSIDADE VEGETAL: ABORDAGEM NO ENSINO MÉDIO

As plantas constituem a base dos ecossistemas e a necessidade de conservar a biodiversidade é imprescindível para a sociedade. O objetivo deste projeto foi conscientizar os alunos de uma escola pública de ensino médio do Rio de Janeiro desta realidade, gerando oportunidade para graduandos e pós-graduandos transmitirem conceitos de biotecnologia aplicada à conservação de plantas. A metodologia consistiu de palestras, documentários em vídeo e visita técnica aos laboratórios do Núcleo de Biotecnologia Vegetal da Universidade do Estado do Rio de Janeiro, onde os alunos acompanharam a produção e a conservação de plantas in vitro. A avaliação baseou-se na frequência e na participação, além da comparação das respostas a um questionário aplicado no início e ao final do projeto. Foi detectada uma grande variação entre as turmas, tanto na frequência (41-68%), quanto no percentual de acertos nos questionários, inicial (42-57%) e final (53-68%). Contudo, todas as turmas avaliadas apresentaram maior rendimento após a participação no projeto, indicando um efeito positivo nas questões específicas de Biotecnologia e sua aplicação na conservação da Biodiversidade. Palavras-chave: Desenvolvimento Sustentável, Cultura In Vitro, Ensino de Biologia, Biotecnologia Vegetal   In vitro conservation of plant biodiversity: approach in high school Abstract: Plants constitute the base of ecosystems and the need to conserve biodiversity is indispensable for society. The aim of this project was to raise awareness among students of a public high school in Rio de Janeiro, creating opportunity for undergraduate and graduate students to transmit concepts of biotechnology applied to plants’ conservation. The methodology consisted of lectures, videos documentaries and a technical visit to laboratories of the Plant Biotechnology Center of the Rio de Janeiro State University where students saw in vitro production and conservation of plants. The evaluation was based on the attendance and participation, as well as the comparison of the answers to questionnaires applied in the beginning and the end of the project. A large variation in the results between the classes was detected both in the frequency (41-68%) and in the percentage of correct answers in the initial (42-57%) and final (53-68%) questionnaires. However, all evaluated classes showed higher performance after participating in the project, indicating a positive effect on specific issues of biotechnology and its application in biodiversity conservation. Keywords: Sustainable Development, In Vitro Culture, Biology Teaching, Plant Biotechnology     Conservación in vitro de biodiversidad vegetal: enfoque en la enseñanza media Resumen: Las plantas constituyen la base de los ecosistemas y la necesidad de conservar la biodiversidad es imprescindible para la sociedad. El objetivo de este proyecto fue concientizar a los alumnos de la enseñanza media pública de esta realidad, generando oportunidad para graduandos y post-graduandos transmitir conceptos de biotecnología aplicada a la conservación de plantas. La metodología consistió de conferencias, vídeos y visita técnica a los laboratorios del Núcleo de Biotecnología Vegetal de la Universidad del Estado de Río de Janeiro, donde los alumnos acompañaron la producción y la conservación de plantas in vitro. La evaluación se basó en la comparación de las respuestas a un cuestionario aplicado al principio y al final del proyecto. Se detectó una gran variación en los resultados entre las clases, tanto en la frecuencia (41-68%), como en el porcentaje de aciertos en los cuestionarios inicial (42-57%) y final (53-68%). Sin embargo, todas las clases evaluadas presentaron mayor rendimiento después de la aplicación en el proyecto, indicando un efecto positivo en las cuestiones específicas de Biotecnología y su aplicación en la conservación de la Biodiversidad. Palabras-clave: Desarrollo Sostenible, Cultivo In Vitro, Enseñanza de Biología, Biotecnología Vegeta

Evaluation of cryopreservation of Petiveria alliacea somatic embryos based on stress caused for the method used / Avaliação da criopreservação de embriões somáticos de Petiveria alliacea com base no estresse causado pelo método usado

Petiveria alliacea is a medicinal species with great potential for pharmacological use against several pathologies, including neoplasms. Many studies have been developed to optimize efficient production methodologies and long-term conservation of this species' genetic resources, with a view to phytochemical and pharmacological research. This study demonstrates the efficiency of the D-cryoplate technique, applied to somatic embryos from plants maintained in vitro. Leaf explants were inoculated in culture medium containing 20 ?M PIC and incubated under standard conditions in plant tissue culture. After 60 days, the somatic embryos induced directly on the leaf tissue surface were transferred to the multiplication medium (MS0). For cryopreservation, samples of these embryos were precultured for 24 hours in medium supplemented with sucrose (0.5M), then groups of 3-5 somatic embryos were encapsulated in calcium chloride directly in aluminum cryoplates. The cryoplates with somatic embryos adsorbed were immersed in a conditioning solution (loading) for 20 min. After being removed from loading, the somatic embryos adhered to the cryoplates were exposed to laminar flow air for different periods of time (0 to 140 min) to assess the level of dehydration. Then, samples submitted at each time were immersed in liquid nitrogen for 2 min. After this time, the cryoplates were removed and kept at room temperature for 20 min, and the somatic embryos were cultivated in MS0 medium. Evaluation after each treatment showed a high survival rate (93%) in cryopreserved somatic embryos. After 90 days of culture it was observed that somatic embryos dehydrated for 120 min showed the highest multiplication rate (32 embryos/inoculated embryo) obtained so far with these explants. The D-cryoplate technique brought innovation to established protocols representing the best option for in vitro conservation of these structures biotechnologically produced that are so promising for phytochemical and pharmacological research

Effects of auriculotherapy on nausea and vomiting in pregnant women:a randomized clinical trial

Background: Pregnancy induces physiological changes, commonly marked by nausea and vomiting in the first trimester, posing risks for both mother and baby. This study evaluates the effects of auriculotherapy on nausea and vomiting during the first trimester of pregnancy.Materials and Methods: A randomized clinical trial was conducted in two primary health care centers with 56 Brazilian pregnant women who reported nausea or vomiting in the first trimester. The participants were divided into an intervention group (auriculotherapy with seeds) and a placebo group (sham auriculotherapy). The intervention was divided into three moments: pre-intervention with assessment of nausea and vomiting and application of questionnaires, and two follow-ups conducted on the fourth and seventh day of the intervention, with reassessment of nausea and vomiting.Results: Both groups experienced a decrease in nausea and vomiting over time, with no statistically significant differences between groups in the within-group analyses at various time points. The intervention group had a greater reduction in symptoms. Within the intervention group, symptoms were more common among ferrous sulfate users and those without reported dietary disturbances. In addition, a higher incidence of nausea and vomiting was associated with the use of analgesics, morning snacks, and low intake of protein, vegetables, and fruits.Conclusions: The intervention did not affect the between-group differences in the incidence of nausea and vomiting and vomiting effort in the first trimester of pregnancy. However, a greater reduction was observed in the intervention group

Micropropagation and evalvation of the genotoxic potential of petiveria alliacea L.

Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorPetiveria alliacea L. é uma planta pertencente à família Phytolaccaceae, conhecida popularmente no país como guiné, erva-de-alho, erva-tipi ou amansa-senhor. Nativa da Região Amazônica tem sido cultivada em muitas áreas tropicais com propósito medicinal ou ritualístico. O objetivo desse trabalho foi (i) o desenvolvimento e a multiplicação de plantas de P. alliacea L. através de métodos de cultura de tecidos, e monitoramento fitoquímico das culturas, e (ii) avaliação comparativa das potencialidades genotóxica e antigenotóxica entre plantas coletadas no campo e produzidas in vitro. Exemplares de diferentes populações ocorrentes no estado do Rio de Janeiro foram utilizados como matrizes para a cultura. Foi estabelecido um protocolo para multiplicação das plantas em meio MS suplementado com BAP e ANA em diferentes concentrações e combinações, que forneceu como melhor resultado em média 8 plantas por explante na concentração de BAP 4,4 μM + ANA 0,54 μM. A análise fitoquímica foi baseada em métodos cromatográficos de diferentes extratos de plantas de campo e plantas in vitro das populações estudadas resultando em diferentes substâncias identificadas nas amostras analisadas por cromatografia em camada delgada. Os extratos foram também avaliados por cromatografia gasosa acoplada á espectrometria de massas, sendo identificadas diferentes substâncias, entre as quais o dibenzil dissulfeto, um produto de degradação de tiosulfinatos com importantes atividades biológicas na defesa das plantas. Os extratos aquosos das plantas de campo e daquelas estabelecidas in vitro foram submetidos à avaliação da potencialidade genotóxica e antigenotóxica, usando-se como modelo o DNA plasmidial pUC 9.1. Os resultados demonstraram que as concentrações utilizadas do extrato aquoso foram capazes de induzir alterações na conformação estrutural do DNA, indicando a ocorrência de quebras simples e duplas nesta molécula. Observou-se também que as lesões aumentaram, proporcionalmente ao aumento da concentração dos extratos, caracterizando-se, assim, um efeito dose-resposta. Os dados também apontaram para um efeito protetor do extrato aquoso, em relação aos danos oxidativos causados pelo cloreto estanoso, indicando, também, uma potencialidade antigenotóxica do extrato aquoso.Petiveria alliacea L. popularly known in Brazil as Guiné, Amansa-senhor or Tipi belongs to the Phytolaccaceae family. Native from the Amazon region, it has been cultivated in many tropical areas for medicinal or ritualistic purposes. The aim of this study was: i) the development and multiplication of plants of P. alliacea L. through tissue culture methods as well as monitoring phytochemical cultures; ii) comparative assessment of genotoxic and antigenotoxic potentialities between field collected and in vitro grown plants. Specimens from different populations occurring in the state of Rio de Janeiro were used as matrices for the culture. It was established a protocol for propagation in MS medium supplemented with different BAP + ANA concentrations and combinations whose best result (BAP 4,4 μM + ANA 0,54 μM) provided 8 plants per explant in average. The phytochemical analysis was based on chromatographic methods of different extracts both of field and in vitro plant populations resulting in different substances identified through the Thin-layer chromatography analyzing samples. The extracts were also assayed by cromatografia gasosa acoplada á espectrometria de massas having identified various substances, including the dibenzyl disulfide, a degradation tiosulfinatos product with important function in plant defense and increasing use by the pharmaceutical industry. Field and in vitro plant aqueous extracts were evaluated for genotoxic and antigenotoxic potentialities using PUC 9.1 plasmid DNA. These results showed changes in the DNA structural conformation indicating the occurrence of single and double strand breaks induced by the extracts in a dose-response way. Finally, data also pointed to a protective effect of aqueous extract against stannous chloride oxidative damage, characterizing an antigenotoxic potentiality of aqueous extract

Avaliação da acurácia altimétrica em levantamentos topográficos urbanos com aeronaves remotamente pilotadas

O mercado de Geotecnologias testemunha o notável aumento da adoção de Aeronaves Remotamente Pilotadas (RPA) em levantamentos topográficos. Este fenômeno é impulsionado por diversos fatores preponderantes, notadamente o substancial aumento na eficiência operacional, com potencial de elevar a produtividade em até 80%, e a consequente redução de custos associados. Nesse contexto, surge a indagação crítica: em aplicações urbanas que requerem levantamentos topográficos de elevada acurácia, as RPAs podem ser empregadas na produção de Informações Geoespaciais? O propósito primordial deste estudo é responder a essa indagação ao avaliar a acurácia altimétrica de RPAs em levantamentos topográficos urbanos, por meio da aplicação do método de controle de qualidade cartográfica estabelecido pelo Decreto-Lei nº 89.817/1984, conhecido como Padrão de Exatidão Cartográfico para Produtos Cartográficos Digitais (PEC-PCD), em conjunto com as Especificações Técnicas de Aquisição de Dados Geoespaciais Vetoriais (ET-ADGV). O levantamento aerofotogramétrico executado com RPAs na área urbana designada é seguido por procedimentos fotogramétricos específicos, e os produtos topográficos resultantes são submetidos à aplicação do Decreto-Lei 89.817 e das ET-ADGV. Nesse contexto, os produtos topográficos, classificados como Classe C na escala 1:1.000 e com equidistância de curvas de nível de 2 metros, são enfatizados neste estudo. Portanto, este trabalho destaca a relevância e a necessidade da aplicação do Decreto-Lei nº 89.817 no controle de qualidade dos produtos topográficos gerados por RPAs, bem como os cuidados técnicos essenciais para alcançar uma classificação mais precisa

Atitudes, crenças e barreiras para a adesão à dieta hipossódica em pacientes com insuficiência cardíaca

Objetivos: Identificar atitudes, crenças e barreiras para a adesão à dieta hipossódica em pacientes com Insuficiência Cardíaca e fatores associados. Métodos: Estudo transversal com 41 indivíduos. Utilizou-se o Questionário de Restrição de Sódio na Dieta. Para análise aplicou-se o teste de Fisher. Resultados: As atitudes basearam-se nas relações familiares e com profissionais; nas crenças destacou-se acreditar na dieta para melhoria da saúde. Como barreiras, encontrou-se empenho insuficiente para mudança dietética e sabor dos alimentos com pouco sal. Associou-se à adesão entre os componentes fatores como: renda, percepção da melhora cardíaca com a dieta, histórico familiar de insuficiência renal e cardíaca e adesão familiar à dieta. Conclusões: As atitudes e crenças identificadas referem-se, respectivamente, às relações sociais e à confiança na dieta para melhorar a condição de saúde. As barreiras foram: empenho insuficiente em modificar a dieta e a palatabilidade alimentar. Fatores econômicos, familiares e clínicos estiveram associados à adesão hipossódica. Palavras-chave: Enfermagem. Insuficiência cardíaca. Dieta hipossódica

Cytotoxicity of Extracts from <i>Petiveria alliacea</i> Leaves on Yeast

Petiveria alliacea L. is a plant used in traditional medicine harboring pharmacological properties with anti-inflammatory, antinociceptive, hypoglycemiant and anesthetic activities. This study assessed the potential cytotoxic, genotoxic and mutagenic effects of ethanolic extract of P. alliacea on Saccharomyces cerevisiae strains. S. cerevisiae FF18733 (wild type) and CD138 (ogg1) strains were exposed to fractioned ethanolic extracts of P. alliacea in different concentrations. Three experimental assays were performed: cellular inactivation, mutagenesis (canavanine resistance system) and loss of mitochondrial function (petites colonies). The chemical analyses revealed a rich extract with phenolic compounds such as protocatechuic acid, cinnamic and catechin epicatechin. A decreased cell viability in wild-type and ogg1 strains was demonstrated. All fractions of the extract exerted a mutagenic effect on the ogg1 strain. Only ethyl acetate and n-butanol fractions increased the rate of petites colonies in the ogg1 strain, but not in the wild-type strain. The results indicate that fractions of mid-polarity of the ethanolic extract, at the studied concentrations, can induce mutagenicity mediated by oxidative lesions in the mitochondrial and genomic genomes of the ogg1-deficient S. cerevisiae strain. These findings indicate that the lesions caused by the fractions of P. alliacea ethanolic extract can be mediated by reactive oxygen species and can reach multiple molecular targets to exert their toxicity

A chloroquinoline derivate presents effective in vitro and in vivo antileishmanial activity against Leishmania species that cause tegumentary and visceral leishmaniasis.

The identification of new therapeutics to treat leishmaniasis is desirable, since available drugs are toxic and present high cost and/or poor availability. Therefore, the discovery of safer, more effective and selective pharmaceutical options is of utmost importance. Efforts towards the development of new candidates based on molecule analogs with known biological functions have been an interesting and cost-effective strategy. In this context, quinoline derivatives have proven to be effective biological activities against distinct diseases. In the present study, a new chloroquinoline derivate, AM1009, was in vitro tested against two Leishmania species that cause leishmaniasis. The present study analyzed the necessary inhibitory concentration to preclude 50% of the Leishmania promastigotes and axenic amastigotes (EC50 value), as well as the inhibitory concentrations to preclude 50% of the murine macrophages and human red blood cells (CC50 and RBC50 values, respectively). In addition, the treatment of infected macrophages and the inhibition of infection using pre-treated parasites were also investigated, as was the mechanism of action of the molecule in L. amazonensis. To investigate the in vivo therapeutic effect, BALB/c mice were infected with L. amazonensis and later treated with AM1009. Parasitological and immunological parameters were also evaluated. Clioquinol, a known antileishmanial quinoline derivate, and amphotericin B (AmpB), were used as molecule and drug controls, respectively. Results in both in vitro and in vivo experiments showed a better and more selective action of AM1009 to kill the in vitro parasites, as well as in treating infected mice, when compared to results obtained using clioquinol or AmpB. AM1009-treated animals presented significantly lower average lesion diameter and parasite burden in the infected tissue and organs evaluated in this study, as well as a more polarized antileishmanial Th1 immune response and low renal and hepatic toxicity. This result suggests that AM1009 should be considered a possible therapeutic target to be evaluated in future studies for treatment against leishmaniasis