Propagação in vitro e aclimatização de Lippia rotundifolia, uma espécie endêmica dos Campos Rupestres Brasileiros

The importance in folk medicine, combined to threats in their environment, becomes necessary to carry out studies involving large-scale propagation of Lippia genus. Although the tissue culture propagation is widely disseminated for medicinal plants, for L. rotundifolia any article was published yet. The present study aimed to establish an efficient protocol for micropropagation of L. rotundifolia. Nodal segments, taken from plants collected in the Espinhaço Range, were disinfected, and cultures were initiated on MS medium with PVPP (1 g L- 1), sucrose (3%) and agar (0.7%). The culture were maintained in a growth room at controlled conditions. Disinfestation procedures and the supply of PVPP on culture media resulted in both reduced contamination and phenol oxidation rates, with more than 90% of viable cultures. In the multiplication phase were tried different BAP and NAA combinations supplied to the MS medium. The treatment that resulted in highest multiplication rates was 0.33 µM BAP. The effects of NAA were evaluated for in vitrorooting. At 0.44 µM, rooting was 70% higher than that observed in the control. The acclimatization was held in trays with substrate, coated with translucent plastic and kept under shade. The plantlets were transferred to the greenhouse after 15 days and transplanted to plant beds after 30 days. The acclimatized plantlets bloomed one year after the transference to field conditions, showing that the in vitro culture did not affect the vegetative and reproductive development, which confirms the potential of micropropagation to reduce the extinction risk of L. rotundifolia.A importância na medicina popular, assim como as ameaças em seu ambiente, tornam necessária a realização de estudos envolvendo a propagação em larga escala de plantas do gênero Lippia. Embora a propagação in vitro seja amplamente disseminada para plantas medicinais, nenhum artigo foi publicado ainda com L. rotundifolia. O presente estudo teve como objetivo estabelecer um protocolo eficiente para a micropropagação dessa espécie. Segmentos nodais, retirados de plantas coletadas na Cadeia do Espinhaço, foram desinfetados antes da inoculação em meio MS suplementado com PVPP (1 g L-1), sacarose (3%) e ágar (0,7%). Os tubos de ensaio foram mantidos em sala de crescimento sob condições controladas. O tratamento de desinfestação e a adição de PVPP aos meios de cultura resultaram na redução da infecção microbiológica assim como nas taxas de oxidação fenólica, proporcionando mais de 90% de culturas assépticas e viáveis . Na fase de multiplicação foram testadas diferentes combinações de BAP e ANA adicionadas ao meio MS. O tratamento que resultou em maiores taxas de multiplicação foi de 0,33 µM de BAP. Os efeitos do ANA foram avaliados no enraizamento in vitro. A 0,44 µM de ANA, o enraizamento foi 70% superior ao observado no controle. A aclimatização foi realizada em bandejas com substrato, revestidas com plástico translúcido e mantidas à sombra. As plantas foram transferidas para casa de vegetação após 15 dias e transplantadas para canteiros após 30 dias. As plantas aclimatizadas floresceram após um ano da transferência para condições de campo, demonstrando que os procedimentos de cultivo in vitro não afetaram o desenvolvimento vegetativo e reprodutivo das plantas, o que confirma o potencial da micropropagação para redução dos riscos de extinção de L. rotundifolia

Nano-zero-valent particles synthesized with agroindustry wastes for pesticide degradation under real conditions

Nano-zero-valent particles (NZVP) had exhibited high degradation activity. NZVP synthesized from agroindustry residues align with circular economy principles. They generate hydroxyl radicals (4.6 µM) that effectively degraded chlorpyrifos-methyl pesticide under real conditions. Bimetallic NZVP, specifically NZVP-Fe:Mn and NZVP-Fe:Ag, show superior pesticide degradation. The metal ratio within NZVP influences their activity (optimal at 0.12:0.12 mM and 0.12:0.19 mM for respectively, NZVP-Fe:Mn and NZVP-Fe:Ag). NZVP characterization includes TEM, SEM-EDS, PZC, FTIR, XRD, and electrochemical analysis, confirming their acid nature, favorable electrochemical behavior, and uniform metal distribution. The impact of different natural extracts on NZVP synthesis and pesticide degradation was explained through extensive extract characterization, revealing the presence of altering pro-oxidants and scavenger species. Blueberry pruning extract yields the highest pesticide degradation (85% in 5 min) due to its stronger antioxidant activity and lower scavenger compound content. NZVP demonstrates efficacy across various pH ranges. Real wastewater samples were treated under optimal conditions, resulting in a pesticide degradation efficiency of approximately 60% within 5 min. The most effective approach for enhancing the treatment process involved the sequential addition of reagents, as opposed to the conventional method of increasing reagent concentrationXunta de Galicia | Ref. ED481B 2019/091Fundação para a Ciência e a Tecnologia | Ref. UIDB/50006/2020Fundação para a Ciência e a Tecnologia | Ref. UIDP/50006/2020Fundação para a Ciência e a Tecnologia | Ref. LA/P/0008/2020Ministerio de Ciencia e Innovación | Ref. PCI2022-132941Universidade de Vigo/CISUGFundação para a Ciência e a Tecnologia | Ref. CEECIND/02702/201

Vena cava thrombosis associated with nephrotic syndrome in the puerperal gestational cycle

CONTEXT: The puerperal gestational cycle is accompanied by a state of physiological hypercoagulability. Thromboembolic phenomena may occur at this time. OBJECTIVE: To report on a clinic case involving a patient that presented a family history of thromboembolism and developed deep vein thrombosis in a lower limb and vena cava thrombosis during the puerperal gestational cycle, displaying nephrotic syndrome as the main complication. DESIGN: Case report.CONTEXTO: A gestação é acompanhada de um estado fisiológico de hipercoagulabilidade. Fenômenos de Trombose podem ocorrer. OBJETIVO: Relatar um caso clínico envolvendo uma paciente que apresentava história importante de trombifilia familiar, e que desenvolveu trombose venosa profunda no membro inferior e trombose da veia cava durante o ciclo gravídico-puerperal, apresentando síndrome nefrótica como principal complicação. TIPO DE ESTUDO: Relato de caso.Universidade de Santo AmaroUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Discipline of RheumatologyUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Head of the Discipline of Urgent MedicineUniversidade de Santo Amaro Discipline of RheumatologyUNIFESP, EPM, Discipline of RheumatologyUNIFESP, EPM, Head of the Discipline of Urgent MedicineSciEL

Estresse oxidativo em plantas micropropagadas de pitcairnia albiflos herb. (bromeliaceae) durante a aclimatização e sob estresse hídrico

Pitcairnia albiflos Herb. (Bromeliaceae) is currently in the list of endangered species. This species is endemic of the inselbergs of the city of Rio de Janeiro, RJ, and has been suffering with the mountaineer’s trampling, wildfires, invasion of exotic grasses and plant extraction. The micropropagation can be used as an alternative to the risk conditions under which populations of this species are submitted, aiming at the recomposition of endangered populations in the natural environment, as well as the supply of the consumer market. The final stage of the micropropagation is the acclimatization, period in which the plants become more susceptible and suffer from oxidative stress due to the changes in the environmental conditions. In the present study it were evaluated the antioxidative enzymatic activities of CAT, SOD, POD, PPO and proline content, besides the levels of photosynthetic pigments in plants of Pitcairnia albiflos grown in vitro in culture mediums containing two sucrose concentrations (15 or 30 g L-1). Part was covered with unsealed lids that allowed gas exchanges and part was kept in sealed flasks with lids and PVC plastic film, that didn’t allow the ventilation. Under these conditions, the plants were cultivated in culture mediums containing GA3 or NAA. After the in vitro growth period, the plants were transferred to ex vitro conditions at a greenhouse. The above mentioned analyses and the determination of total soluble carbohydrate levels, sucrose, starch, reducer sugars, relative water content and succulence were also performed on the plants previously grown in vitro with 15 or 30 g L-1 of sucrose and GA3 in tubes with sealed lids, after the submission of these to water stress during 24, 38 or 52 days. After the water stress period, the plants were rehydrated for 34 days under regular irrigation at the greenhouse. In the in vitro cultivated tissues it was noted the emergence of hyperhydricity characteristics in the plants grown with 15 g L-1 of sucrose, GA3 and tubes with sealed lids, which was evidenced by the lowest proline accumulation, the increased in the antioxidative enzymatic activities and the lowest accumulation of photosynthetic pigments. In the ex vitro condition, the plants previously grown in culture mediums containing 15 g L-1 of sucrose presented larger antioxidative enzymatic activities, which did not show, in some cases, significant differences compared with the largest concentration of sucrose. In this condition, the proline accumulation was lower, which is an indicative of larger oxidative stress in these plants during acclimatization. During the water stress, there was a fall in the activity of all studied enzymes, although that fall had been more evident in the plants that were initially cultivated with 15 g L-1 of sucrose. The proline accumulation increased with the extension of the water stress, being larger in the plants grown in vitro with 30 g L-1 of sucrose. There were no viii significant differences in the content of photosynthetic pigments and in their relation with any sucrose concentrations, except for total carotenoids, which significantly increased over the period of water stress for the plants previously grown with the lowest concentration of sucrose. The contents of total soluble carbohydrates and sucrose increased with the extension of the stress, being more accentuated in the 30 g L-1 of sucrose concentration. The contents of starch, the relative content of water and succulence presented a reduction with the increase of water stress. After the rehydration, all plants showed recovery capacity, presenting values close to those from the control groups for all the analyzed variables. It should be noted, however, that the plants treated with 30 g L-1 of sucrose had better recovery compared with those that were treated with 15 g L-1 of sucrose. According to the obtained results, it is possible to conclude that the concentration of sucrose used in vitro presents influence on the process of ex vitro acclimatization and also, later in the field, in the recovery capacity of the plants to drought when they are submitted to water stress. The plants grown in vitro with 15 g L-1 of sucrose were more sensitive to drought and, possibly, would not survive if they were transferred from the test tubes directly to the field. The plants grown in vitro with 30 g L-1 of sucrose were apparently more resistant to the ex vitro acclimatization process, presenting greater survival chances in the field, larger drought tolerance and larger recovery capacity after extended periods of water stress.Pitcairnia albiflos Herb. (Bromeliaceae) atualmente se encontra na lista de espécies ameaçadas de extinção. Essa espécie é endêmica dos afloramentos rochosos do município do Rio de Janeiro, RJ, e vem sofrendo com o pisoteio de alpinistas, queimadas, invasão de gramíneas exóticas e extrativismo vegetal. A micropropagação pode ser utilizada como alternativa às condições de risco em que as populações dessa espécie se encontram submetidas, visando à recomposição de populações ameaçadas em ambiente natural, assim como o abastecimento do mercado consumidor. A etapa final da micropropagação é a aclimatização, período em que as plantas ficam mais susceptíveis e sofrem com o estresse oxidativo devido às mudanças nas condições ambientais. No presente trabalho foram avaliadas as atividades enzimáticas antioxidativas da CAT, SOD, POD, PPO e o conteúdo de prolina, além dos teores de pigmentos fotossintéticos em plantas de Pitcairnia albiflos cultivados in vitro, em meios de cultura contendo duas concentrações de sacarose (15 ou 30 g L-1), tampas não vedadas que permitiam trocas gasosas e frascos vedados com tampas e filme plástico de PVC, que impediam a ventilação. Sob essas condições, as plantas foram cultivadas em meios contendo GA3 ou ANA. Após o período de crescimento in vitro, as plantas foram transferidas para condições ex vitro em casa de vegetação. As análises supra-citadas e a determinação dos teores de carboidratos solúveis totais, sacarose, amido, açúcares redutores, conteúdo relativo de água e suculência também foram realizadas nas plantas previamente cultivadas in vitro com 15 ou 30 g L-1 de sacarose e GA3 em tubos com tampas vedadas, após submissão das mesmas a estresse hídrico durante 24, 38 ou 52 dias. Após o período de estresse hídrico, as plantas foram reidratadas durante 34 dias sob irrigação periódica em casa de vegetação. Nos tecidos cultivados in vitro percebeu-se o surgimento de características de hiperidricidade nas plantas cultivadas com 15 g L-1 de sacarose, GA3 e tubos com tampas vedadas, o que foi evidenciado pelo menor acúmulo de prolina, aumento das atividades das enzimas antioxidantes e menor acúmulo de pigmentos fotossintetizantes. Na condição ex vitro, as plantas cultivadas anteriormente em meio de cultura contendo 15 g L-1 de sacarose apresentaram maior atividade das enzimas antioxidantes não havendo, em alguns casos, diferenças significativas em comparação com a concentração mais elevada de sacarose. Nessa condição o acúmulo de prolina foi menor, o que é indicativo de maior estresse oxidativo nessas plantas durante a aclimatização. Durante o estresse hídrico houve queda na atividade de todas as enzimas estudadas, embora essa queda tenha sido mais acentuada para as plantas que inicialmente foram cultivadas com 15 g L-1 de vi sacarose. O acúmulo de prolina aumentou com o prolongamento do estresse hídrico, sendo maior nas plantas que foram cultivadas in vitro com 30 g L-1 de sacarose. Não houve diferenças significativas no conteúdo de pigmentos fotossintetizantes e nas suas relações para nenhuma das concentrações de sacarose, exceto para os carotenóides totais, que apresentaram aumento significativo ao longo do período de estresse hídrico para as plantas previamente cultivadas com a menor concentração de sacarose. Os conteúdos de carboidratos solúveis totais e sacarose aumentaram com o prolongamento do estresse, sendo mais acentuados na concentração de 30 g L-1 de sacarose. Os conteúdos de amido, o conteúdo relativo de água e a suculência apresentaram redução com o aumento do estresse hídrico. Após a reidratação, todas as plantas mostraram capacidade de recuperação, apresentando valores próximos aos dos controles para todas as variáveis analisadas. Ressalta-se, todavia, que as plantas tratadas com 30 g L-1 de sacarose tiveram melhor recuperação quando comparadas com aquelas que foram tratadas com 15 g L-1 de sacarose. Em função dos resultados obtidos, é possível concluir que a concentração de sacarose utilizada in vitro apresenta influência no processo de aclimatização ex vitro e também, posteriormente, no campo, na capacidade de recuperação das plantas à seca quando elas são submetidas a estresse hídrico. As plantas cultivadas in vitro com 15 g L-1 de sacarose se mostraram mais sensíveis à seca e, possivelmente, não sobreviveriam caso fossem transferidas dos tubos de ensaio diretamente para o campo. As plantas cultivadas in vitro com 30 g L-1 de sacarose aparentemente eram mais resistentes ao processo de aclimatização ex vitro, apresentando maiores chances de sobrevivência em campo, maior tolerância à seca e maior capacidade de recuperação após períodos prolongados de estresse hídrico

In vitro propagation and acclimatization of Lippia rotundifolia, an endemic species of Brazilian Campos Rupestres

ABSTRACTThe importance in folk medicine, combined to threats in their environment, becomes necessary to carry out studies involving large-scale propagation of Lippia genus. Although the tissue culture propagation is widely disseminated for medicinal plants, for L. rotundifolia any article was published yet. The present study aimed to establish an efficient protocol for micropropagation of L. rotundifolia. Nodal segments, taken from plants collected in the Espinhaço Range, were disinfected, and cultures were initiated on MS medium with PVPP (1 g L- 1), sucrose (3%) and agar (0.7%). The culture were maintained in a growth room at controlled conditions. Disinfestation procedures and the supply of PVPP on culture media resulted in both reduced contamination and phenol oxidation rates, with more than 90% of viable cultures. In the multiplication phase were tried different BAP and NAA combinations supplied to the MS medium. The treatment that resulted in highest multiplication rates was 0.33 µM BAP. The effects of NAA were evaluated for in vitrorooting. At 0.44 µM, rooting was 70% higher than that observed in the control. The acclimatization was held in trays with substrate, coated with translucent plastic and kept under shade. The plantlets were transferred to the greenhouse after 15 days and transplanted to plant beds after 30 days. The acclimatized plantlets bloomed one year after the transference to field conditions, showing that the in vitro culture did not affect the vegetative and reproductive development, which confirms the potential of micropropagation to reduce the extinction risk of L. rotundifolia

MICROPROPAGATION AND ACCLIMATIZATION OF Aegiphila verticillata Vell.: AN ENDANGERED WOODY SPECIES

The objective of this work was to establish an efficient protocol for in vitro multiplication and rooting, as well as ex vitro acclimatization of Aegiphila verticillata, a woody species found in Brazilian rocky fields. Aseptic cultures were established by seeds and two multiplication analyses were performed. In the first, we employed 6-benzylaminopurine (BAP – 0, 2.5, 5 and 7.5 μM) + α-naphthalene acetic acid (NAA – 0, 0.2, 0.4 and 0.6 μM) and, in the second, were studied adenine sulfate, kinetin and thidiazuron (0, 5, 7.5, 10 and 12.5 μM). After 90 days, we assessed the quantitative and qualitative shoot propagation. There were more than 90% seed germination and low contamination (2%). In multiplication phase, the culture medium that promoted the best quantitative and qualitative culture development was supplemented with 7.5 μM BAP + 0.4 μM NAA. In the rooting assay, were used NAA, indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) (0, 0.1, 0.2, 0.3 or 0.4 μM). After 90 days, the root number and rooting quality were evaluated. In this analysis, differences were not found between the control and the other treatments. Rooted plantlets were acclimatized in styrofoam trays for 30 days, after which they were transferred to pots in the greenhouse. Only 3% of the plants subjected to initial acclimatization died and 70% of the plants transferred to the field conditions survived and showed normal development. The results founded in this work are the first involving in vitro propagation and ex vitro acclimatization of Aegiphila verticillata and provide a continuous supply of this medicinal native species, endangered due anthropogenic activities