Cytotoxicity of isolated compounds from Picrasma crenata (Vell) Engl. in animal tumor cell (HTC) / Citotoxicidade de compostos isolados de Picrasma crenata (Vell) Engl. em células tumorais animais (HTC)

The study aim was to evaluate the cytotoxic activity, using the MTT test [3-(4,5-Dimethilthiazol-2-yl)-2,5-diphenil tetrazolium bromide], from the crude extract of Picrasma crenata (Pau Tenente) and its isolated compounds, quassin and parain, in culture of rat liver tumor cells (HTC). The test was carried out exposing the cells for 24, 48 and 72 hours to concentrations of 5, 10, 50, 100, 200, 300, 400, 500 and 1000 µg of crude extract of Pau Tenente/mL of culture medium and 1, 5, 10, 15, 20, 40, 60, 80 and 100 µg of quassin or parain compounds/mL of culture medium. The absorbances averages results obtained showed that the crude extract did not present cytotoxicity for the HTC cells in all the concentrations and evaluated times. For quassin, the concentrations of 80 and 100 µg/mL were cytotoxic, after 72 hours of treatment. For parain, the concentrations of 1, 5, 20, 40, 60, 80 and 100 µg/mL, in 72 hours, were cytotoxic, revealing a new activity for this compound. Thus, the results demonstrate a first indication of the cytotoxic activity of compounds quassin and parain, adding an important social and economic value to them, and may have application in future research and in pharmaceutical industry.

O objetivo do estudo foi avaliar a atividade citotóxica, por meio do teste MTT [brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazólio], do extrato bruto de Picrasma crenata (Pau Tenente) e seus compostos isolados, quassina e paraína, em cultura de células tumorais de fígado de rato (HTC). O teste foi realizado expondo as células por 24, 48 e 72 horas às concentrações de 5, 10, 50, 100, 200, 300, 400, 500 e 1000 µg de extrato bruto de Pau Tenente/mL de meio de cultura e 1, 5, 10, 15, 20, 40, 60, 80 e 100 µg de compostos de quassina ou paraína/mL de meio de cultura. Os resultados das médias de absorbâncias obtidos mostraram que o extrato bruto não apresentou citotoxicidade para as células HTC em todas as concentrações e tempos avaliados. Para quassina, as concentrações de 80 e 100 µg/mL foram citotóxicas, após 72 horas de tratamento. Para a paraína, as concentrações de 1, 5, 20, 40, 60, 80 e 100 µg/mL, em 72 horas, foram citotóxicas, revelando uma nova atividade para este composto. Assim, os resultados demonstram uma primeira indicação da atividade citotóxica dos compostos quassina e paraína, agregando-lhes importante valor social e econômico, podendo ter aplicação em pesquisas futuras e na indústria farmacêutica.

Astragaloside IV alleviates stroke-triggered early brain injury by modulating neuroinflammation and ferroptosis via the Nrf2/HO-1 signaling pathway

Purpose: Stroke is an acute cerebrovascular disease. Astragaloside IV (AS-IV) is an active ingredient extracted from Astragalus membranaceus with an established therapeutic effect on central nervous system diseases. This study examined the neuroprotective properties and possible mechanisms of AS-IV in stroke-triggered early brain injury (EBI) in a rat transient middle cerebral artery occlusion (MCAO) model. Methods: The neurological scores and brain water content were analyzed. 2,3,5-triphenyl tetrazolium chloride (TTC) staining was utilized to determine the infarct volume, neuroinflammatory cytokine levels, and ferroptosis-related genes and proteins, and neuronal damage and molecular mechanisms were evaluated by terminal deoxynucleotidyl transferase dutp nickend labeling (TUNEL) staining, western blotting, and real-time polymerase chain reaction. Results: AS-IV administration decreased the infarct volume, brain edema, neurological deficits, and inflammatory cytokines TNF-α, interleukin-1ß (IL-1ß), IL-6, and NF-κB, increased the levels of SLC7A11 and glutathione peroxidase 4 (GPX4), decreased lipid reactive oxygen species (ROS) levels, and prevented neuronal ferroptosis. Meanwhile, AS-IV triggered the Nrf2/HO-1 signaling pathway and alleviated ferroptosis due to the induction of stroke. Conclusion: Hence, the findings of this research illustrate that AS-IV administration can improve delayed ischemic neurological deficits and decrease neuronal death by modulating nuroinflammation and ferroptosis via the Nrf2/HO-1 signaling pathway.

Suitability of tetrazolium test for Tamarindus indica L. seeds / Adequação do teste de tetrazolium para sementes de Tamarindus indica L

Seed quality is routinely assessed by direct tests, e.g, the germination test, or indirect tests like the tetrazolium test, which has shoown to be promising in the determine viability and vigor, allowing the diagnosis of the main problems that may affect seed quality, such as mechanic damages, field deterioration and storage. In this respect, this study was conducted to develop a tetrazolium test protocol to evaluate the viability and vigor of Tamarindus indica L. seeds. Before exposing the seeds to the tetrazolium solution, seed preconditioning studies were carried out in which seven soaking times were tested. The soaking time that did not cause damage to the seed embryo and allowed the removal of the seed coat to expose the seed structures to the tetrazolium salt was selected. Then, an experiment was set up in a completely randomized design with a 2x3x3 factorial arrangement involving two seed lots, three soaking times in tetrazolium salt (6, 12 and 16 h) and three salt concentrations (0.075, 0.1 and 0.5%), totaling 18 treatments with four replicates of 25 seeds, evaluated at 40 ºC. For each treatment, the seeds were divided into three classes, namely, viable and vigorous embryos (class 1); viable embryos (class 2) and non-viable or dead embryos (class 3). For a comparison with the tetrazolium test results, the two seed lots were analyzed for water content, germination, emergence, first count, germination speed index, emergence speed index, growth and seedling dry weight. The viability and vigor of T. indica seeds can be evaluated after a soaking period of 48 h and immersion for 6h in tetrazolium salt at the concentration of 0.1%, at 40°C, with provides results similar to conventional seed viability tests. The tetrazolium test proved to be adequate to differentiate T. indica seed lots in terms of viability.(AU)

A qualidade das sementes é avaliada rotineiramente por testes diretos como o de germinação e indiretos como o teste de tetrazólio que tem se mostrado promissor na determinação da viabilidade e vigor possibilitando o diagnóstico dos principais problemas que podem afetar a qualidade das sementes, como danos mecânicos, deterioração de campo e armazenamento. Neste sentido, objetivou-se com este trabalho desenvolver um protocolo do teste de tetrazólio para avaliar a viabilidade e o vigor das sementes de Tamarindus indica L. Antes da exposição das sementes a solução de tetrazólio, foram realizados estudos pre-condicionamento das sementes utilizando sete períodos de embebição sendo selecionado o tempo que não causou danos ao embrião da semente e permitiu a remoção do tegumento para exposição das estruturas da semente ao sal de tetrazólio. Em seguida, o experimento foi instalado em delineamento inteiramente casualizado em esquema fatorial 2x3x3, sendo dois lotes de sementes, três tempos de embebição em sal (6; 12 e 16 horas) e três concentrações de sal de tetrazólio (0,075; 0,1 e 0,5%), totalizando dezoito tratamentos com quatro repetições de 25 sementes, avaliados a 40 ºC. Para cada tratamento as sementes foram divididas em três classes: embriões viáveis e vigorosos (classe 1); embriões viáveis (classe 2) e embriões inviáveis ou mortos (classe 3). Para comparar com o resultado do teste de tetrazólio os dois lotes de sementes foram submetidos a análise de teor de água, germinação, emergência, primeira contagem e índice de velocidade de germinação e emergência, crescimento e massa seca de plântulas. A viabilidade e o vigor das sementes de T. indica podem ser avaliados após um período de 48h de embebição e 6h de imersão na concentração de 0,1% de sal de tetrazólio a 40 ºC, com resultados semelhantes aos testes convencionais de determinação da viabilidade das sementes. O teste de tetrazólio mostrou-se adequado para diferenciar a viabilidade em relação aos lotes de sementes de T. indica.(AU)

Antioxidant and physiological responses of seeds of soybean cultivars to delayed harvest / Respostas antioxidantes e fisiológicas de sementes de cultivares de soja ao atraso de colheita

Soybean seeds are susceptible to field deterioration after reaching physiological maturity, which usually occurs at the R7 stage, with harvest at R8. This study aimed to evaluate the biochemical and physiological changes in seeds of different soybean cultivars submitted to delayed harvest in the field. Seeds of the cultivars NS 5959, BMX Potência, and TMG 1175 were harvested at four times (0, 10, 20, and 30 days after reaching R8). Moisture, germination, first count, accelerated aging, emergence, and tetrazolium tests were performed in each harvest period. The activities of the antioxidant enzymes catalase (CAT), peroxidase (POX), and ascorbate peroxidase (APX) were also evaluated. The experiment was carried out in a completely randomized design in a 3×4 factorial scheme (three cultivars and four harvest times). A reduction in germination and vigor of seeds of different soybean cultivars was found at R8 + 20 and R8 + 30 days related to field deterioration due to delayed harvest. The seeds of the cultivars did not differ in terms of viability under delayed harvest but differed in terms of antioxidant and physiological responses. Better and worse seed performances were found for the cultivars BMX Potência and TMG 1175, respectively, at R8 and in R8 + 10 days.(AU)

Sementes de soja são suscetíveis à deterioração em campo após atingirem a maturidade fisiológica, que em geral, ocorre no estádio R7 sendo a colheita realizada em R8. Objetivou-se com esse trabalho avaliar as alterações bioquímicas e fisiológicas em sementes de diferentes cultivares de soja submetidas ao atraso de colheita no campo. Sementes das cultivares NS 5959, BMX Potência e TMG 1175 foram colhidas em quatro épocas (0, 10, 20 e 30 dias após atingirem R8). Em cada período de colheita, foram realizados os testes de umidade, germinação, primeira contagem, envelhecimento acelerado, emergência e tetrazólio. Também foram avaliadas as atividades das enzimas antioxidantes catalase (CAT), peroxidase (POX) e ascorbato peroxidase (APX). O experimento foi conduzido em DIC, em esquema fatorial 3 x 4 (três cultivares e quatro épocas de colheita). Em R8 + 20 e R8 + 30 dias, foi encontrada redução na germinação e vigor das sementes das diferentes cultivares de soja devido à deterioração em campo pelo atraso de colheita. Sob atraso de colheita, as sementes das cultivares não diferiram quanto à viabilidade, mas diferiram quanto às respostas antioxidantes e fisiológicas. No estádio R8 e em R8 + 10 dias foi possível observar melhor desempenho das sementes da cv. 'BMX Potência' e pior para a cv. 'TMG 1175'.(AU)

Physiological and sanitary quality of ryegrass seeds submitted to different defoliation frequencies / Qualidade fisiológica e sanitária de sementes de azevém-anual submetidas a diferentes frequências de desfolha

This study determined the effect of different defoliation frequencies in a long-cycle ryegrass cultivar on yield components and the physiological and sanitary quality of seeds. Four defoliation frequencies were applied (without, one, two, and three defoliations). The time between defoliations was 350 degree-days. At the first, second, and third defoliation time, the plants were 15 cm, 20 cm, and 35 cm, respectively, and were lowered to half their height. The yield components and plant structure were evaluated through the tillers, along with the physiological quality of the seeds, which was verified by germination tests, first germination count, accelerated aging, tetrazolium test, field emergence, and weight of a thousand seeds. For the sanitary quality analysis, the percentage (%) of fungi incidence in the seeds was verified. The second defoliation determined the seeds' high physiological and sanitary quality due to the higher amount of primary and secondary tillers produced concerning the other treatments. However, there was no significant difference for the third defoliation since the aerial tillers issued the same quality of seeds originated from primary and secondary tillers. The average incidence of fungi such as Colletotrichum sp., Fusarium sp., Phoma sp., and Dreshslera spp. was lower in ryegrass plants subjected to three defoliations.

O presente estudo teve como objetivo determinar o efeito de diferentes frequências de desfolha em uma cultivar de azevém de ciclo longo, sobre os componentes de rendimento e a qualidade fisiológica e sanitária das sementes. Foram aplicadas quatro frequências de desfolhas (sem, uma, duas e três). O tempo entre desfolhas foi de 350 graus-dia. No momento da primeira, segunda e terceira desfolha as plantas estavam com 15 cm, 20 cm e 35 cm, respectivamente e foram rebaixadas para metade de suas alturas. Avaliaram-se os componentes de rendimento e a estrutura das plantas por meio dos perfilhos, juntamente com a qualidade fisiológica das sementes, a qual foi verificada por testes de germinação, primeira contagem de germinação, envelhecimento acelerado, teste de tetrazólio, emergência a campo e peso de mil sementes. Para a análise da qualidade sanitária verificou-se a porcentagem (%) de incidência de fungos nas sementes. A segunda desfolha determinou alta qualidade fisiológica e sanitária das sementes devido a maior quantidade de perfilhos primários e secundários produzidos em relação aos demais tratamentos. No entanto não houve diferença significativa para a terceira desfolha, visto que nos perfilhos aéreos emitidos observou-se a mesma qualidade das sementes originadas de perfilhos primários e secundários. A incidência média de fungos como Colletotrichum sp., Fusarium sp., Phoma sp. e Dreshsleras pp. foi menor em plantas de azevém submetidas a três desfolhas.

Recycled pequi (Caryocar brasiliense, Camb) shell ethanolic extract induces apoptosis in canine osteosarcoma cell / Extrato etanólico de cascas recicladas de pequi (Caryocar brasiliense, Camb) induz apoptose em células de osteossarcoma canino

Osteosarcoma is a mesenchymal cancer associated with high mortality in dogs and in humans. The biodiversity-rich Cerrado, the predominant biome in the Midwest region of Brazil, is native to locally appreciated fruits such as pequi (Caryocar brasiliense, Camb). Although this plant has been frequently used in folk medicine, the pharmacological properties of pequi fruit shells have not been fully evaluated. Thus, this study aimed to determine the cytotoxic activity of ethanol extract of recycled pequi shells on canine osteosarcoma cells in vitro. Cells were cultured and treated with final extract concentrations of 0, 0.029 µg/µL, 0.29 µg/µL, and 2.91 µg/µL for 24, 48, or 72 hours. Cell viability assay using trypan blue exclusion method and tetrazolium reduction method, cell survival assay, and double labeling with annexin V and propidium iodide were performed in the treated osteosarcoma cells. These allowed the determination of IC50, survival fraction, and type of cell death, respectively. Pequi shell ethanol extract at a concentration of 2.91 µg/µL showed the greatest inhibition of osteosarcoma cell growth in vitro, resulting in a 71.80% decrease in growth compared to the control. The mean IC50 was 155.2 µg/mL at 72 hours. The calculated survival fractions showed that cell growth at 72 hours was 3.33% lower in cells treated with 2.91 µg/µL extract. Results from the double labeling experiment suggest that apoptosis was the predominant type of cell death in cells treated with 2.91 µg/µL extract. These results demonstrate that ethanol extract of recycled pequi shells promotes apoptosis in canine osteosarcoma cells.

O osteossarcoma é uma neoplasia mesenquimal associada à alta mortalidade em cães e humanos. O pequi (Caryocar brasiliense, Camb.) é nativo do Cerrado, bioma rico em biodiversidade predominante na região Centro-Oeste do Brasil. Embora esta planta seja frequentemente utilizada na medicina popular, as propriedades farmacológicas da casca do pequi ainda são pouco conhecidas. Este estudo teve como objetivo determinar a atividade citotóxica do extrato etanólico de cascas de pequi recicladas em células de osteossarcoma canino. As células foram cultivadas e tratadas com concentrações finais de extrato de zero, 0,029 µg/µL, 0,29 µg/µL e 2,91 µg/µL por 24, 48 ou 72 horas. As células foram submetidas a ensaio de viabilidade celular pelos métodos de exclusão de azul de tripano da redução de tetrazólio para determinar a IC50, ao ensaio de sobrevivência celular para calcular a fração de sobrevivência e à dupla marcação com anexina V e iodeto de propídio para determinar o tipo de morte celular. O extrato etanólico de casca de pequi na concentração de 2,91 µg/µL apresentou a maior inibição do crescimento de células de osteossarcoma, com redução de 71,80% no crescimento em relação ao controle. A IC50 média foi de 155,2 µg/mL em 72 horas. O crescimento celular em 72 horas foi 3,33% menor nas células tratadas com 2,91 µg/µL de extrato. A apoptose foi o tipo predominante de morte celular em células tratadas com 2,91 µg/µL de extrato. Esses resultados demonstram que o extrato etanólico de cascas de pequi recicladas promove apoptose em células de osteossarcoma canino.

Turmeric ethanol extract (Curcuma longa L. ) reduces apoptosis and promotes canine osteosarcoma cell proliferation / Extrato etanólico de açafrão (Curcuma longa L. ) reduz apoptose e promove proliferação de células de osteossarcoma canino

Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic properties, including antineoplastic action. The ethanolic extract of the plant contains several phenolic compounds, especially curcumin. Osteosarcoma is a predominant bone tumor in dogs and humans, characterized by high metastatic potential and an unfavorable prognosis. The aim of this study was to investigate the effects of turmeric ethanol extract on canine osteosarcoma cells from established culture. The cells were cultured and treated with different curcumin concentrations (0, 10 µM, 20 µM, 50 µM, 100 µM, and 1000 µM) and exposure times (24h, 48h, and 72h). We first performed tetrazolium reduction technique (MTT) assay and calculated IC50. An immunocytochemistry assay was performed after extract treatment to verify the expression of mutated p53 and therefore study the proliferative potential of malignant cells; Bcl-2 and Ki-67 were used to assess apoptosis and the degree of malignancy, respectively. The extract enhanced the proliferation of canine osteosarcoma cells, reaching 3,819.74% at 50 µM of curcumin. The extract also significantly altered the expression of mutated p53 and Ki-67 proteins but not that of Bcl-2, suggesting that it did not induce this antiapoptotic pathway. Overall, these results are prerequisite to better understanding how natural compounds such as turmeric ethanolic extract affect cell proliferation and could be used to treat various diseases.

A Curcuma longa L., planta conhecida popularmente como açafrão, tem sido amplamente estudada por suas diversas propriedades terapêuticas, incluindo a ação antineoplásica. O extrato etanólico da planta contém diversos compostos fenólicos, com destaque para a curcumina. O osteossarcoma é um tumor ósseo predominante em cães e humanos, caracterizado por apresentar alto potencial metastático e prognóstico desfavorável. Procurou-se investigar os efeitos de diferentes concentrações de curcumina do extrato etanólico de açafrão sobre células de osteossarcoma canino de cultura estabelecida. As células foram cultivadas e submetidas ao tratamento com extrato com diferentes concentrações de curcumina (0, 10 µM, 20 µM, 50 µM, 100 µM e 1000 µM) e tempos de exposição (24h, 48h e 72h) pelo EEA. Inicialmente, foram realizados: técnica de redução do tetrazólio (MTT) e cálculo da IC50. Posteriormente, após o tratamento com o extrato, realizou-se o ensaio de imunocitoquímica para verificar a expressão de p53 mutada e estudar o potencial proliferativo das células malignas; Bcl-2, com intuito de averiguar o estímulo de via antiapoptótica; e o marcador Ki-67, que sinaliza aumento no grau de malignidade. O extrato promoveu proliferação de células de osteossarcoma canino, com incremento de até 3819,74% na concentração de 50µM de curcumina. O composto também alterou a expressão das proteínas p53 mutante e Ki-67 significativamente, mas não alterou a expressão de Bcl-2, mostrando que não induziu a via antiapoptótica mediada por esta. Estes resultados demonstram que o extrato etanólico do açafrão apresenta potencial proliferativo sobre células de osteossarcoma canino, sugerindo a necessidade de conscientização e conhecimento dos reais efeitos de determinados compostos naturais, considerados seguros ao serem utilizados como tratamento de diversas enfermidades.

Methodology of the tetrazolium test for identifying viable seeds of orchids of the genus Cattleya native to the Brazilian Cerrado / Metodologia do teste de tetrazólio para identificação de sementes viáveis de orquídeas do gênero Cattleya nativas do Cerrado brasileiro

Orchids need measures for conservation of their species, among them the creation of seed banks. This study analyzes suitable methodologies to identify the viability of Cattleya seeds through the tetrazolium test. Seeds of Cattleya nobilior Rchb. and Cattleya walkeriana Gardn. were submitted to three preconditioning: no soaking (control); sucrose (10%); or distilled water. After 24 hours, they were submitted to tetrazolium solution with three types of conditioning: oven (40 °C); water bath (40 °C); or room temperature. Subsequently, the percentage of viable seeds (VS) was calculated and in vitro sowing was carried out. Germination percentage (G) was determined after 45 days. A completely randomized design was used for each species studied, with treatments arranged in a 3x3 factorial scheme (three preconditioning and three conditioning), with eight replicates of one tube each. The experiment was performed in duplicate. The highest viability values found in the tetrazolium test were close to those of germination (C. nobilior: VS = 88% e G = 97%; C. walkeriana: VS = 88% e G = 95%) for Cattleya species native to the Cerrado. The conditioning at ambient temperature had a greater effect on seed visualization for both species studied, suggesting that the tetrazolium test should be performed at this conditioning temperature.(AU)

As orquídeas necessitam de medidas para a conservação de suas espécies, entre essas a criação de bancos de sementes. Objetivou-se identificar metodologia adequada para a identificação da viabilidade de sementes de Cattleya por meio do teste de tetrazólio. Sementes de Cattleya nobilior Rchb. e Cattleya walkeriana Gardn. foram submetidas a três pré-condicionamentos: sem embebição (controle); sacarose (10%) ou água destilada. Decorridas 24 horas, foram submetidas à solução de tetrazólio, com três tipos de condicionamento: estufa (40 °C), banho maria (40 °C) ou temperatura ambiente. Após esse período foi calculada a porcentagem de sementes viáveis (SV). Foi realizada a semeadura in vitro e, após 45 dias, determinada a porcentagem de germinação (G). Para cada espécie estudada, foi utilizado o delineamento inteiramente casualizado e os tratamentos foram arranjados em esquema fatorial 3x3 (três précondicionamentos e três condicionamentos) com oito repetições de um tubo de ensaio cada, sendo o experimento realizado em duplicata. Os maiores valores de viabilidade encontrados no teste de tetrazólio foram próximos aos de germinação (C. nobilior: SV = 88% e G = 97%; C. walkeriana: SV = 88% e G = 95%) para essas espécies de Cattleya nativas do Cerrado. Para as duas espécies estudadas, o condicionamento em temperatura ambiente apresentou melhor visualização das sementes, sugerindo que o teste de tetrazólio deve ser realizado nesta temperatura de condicionamento.(AU)

Weathering deterioration in pre-harvest of soybean seeds: physiological, physical, and morpho-anatomical changes

ABSTRACT: Successive cycles of water absorption and loss favor weathering deterioration, one of the main factors that affect the quality of soybean seeds. This study evaluated the physiological, physical, and morpho-anatomical changes in soybean seeds under weathering deterioration at the pre-harvest phase. Six soybean cultivars (BMX Apolo, DM 6563, NS 5959, NA 5909, BMX Potência, and TMG 1175) were produced in a greenhouse and underwent weathering deterioration through a rainfall simulation system, applying 0, 60, 120, and 180 mm of precipitation at pre-harvest phase. Each rainfall level was divided into two applications at an interval of 72 h: 60 mm (30 + 30), 120 mm (60 + 60), and 180 mm (90 + 90). After harvest, the seeds were evaluated for germination, vigor, physical and morpho-anatomical properties. Weathering deterioration induced by simulated rainfall at the pre-harvest phase contributes to the reduction in soybean seed germination and vigor and is conditioned by the soybean genotype. The increase in intensity of simulated rainfall led to a more significant weathering damage in seeds, as evidenced by the X-ray and tetrazolium test. Cultivars DM 6563 and BMX Potência were more susceptible, while NA 5909 was less susceptible to weathering deterioration (especially at the highest level; 120 mm and 180 mm). Anatomical changes caused by weathering deterioration lead to cell compaction and rupture, mainly in the cell layers of the hourglass and parenchyma, forming intracellular spaces. The presence of weathering damage caused a reduction in physiological soybean seed quality.

Effect of bentonite on the health and dairy production of cows submitted to a diet naturally contaminated by mycotoxins / Efeito da bentonita na saúde e produção leiteira de vacas submetidas à dieta naturalmente contaminada por micotoxinas

Mycotoxins often contaminate cattle food, which can cause liver damage, immunosuppression, and reduced milk production. Although previous studies have shown the benefits of adsorbents in farm animals, knowledge regarding their mechanism of action is limited, especially when intoxication occurs due to naturally contaminated diets. The present study aimed to assess whether the daily oral administration of mycotoxin adsorbent bentonite clay based on aluminosilicate for 56 days, would attenuate these changes in 18 dairy cows, which were multiparous in the middle of the lactation stage, and were consuming a diet containing fumonisin B1 and B2, zearalenone, and desoxynivalenol. The animals were divided into treatment (TG, n = 9) and control (CG, n = 9) groups, and subjected to assessment of liver functions, hematological assessments, assessment of oxidative leucocyte metabolism by the tetrazolium nitroazul (NBT) technique, and physical chemical analysis of milk, every week for two months, totaling eight analyses. It was observed that the use of the adsorbent caused a reduction in the milk excretion of aflatoxin M1 (AFM1), an increase in levels of serum protein (p = 0.03) and albumin (p = 0.0001), an increase in leukocyte oxidative metabolism from day 24 of treatment(p = 0.05), and increased milk production from the day 16 of treatment (p = 0.08). There was no improvement in the physicochemical indices of the milk. It was concluded that the use of an aluminosilicate-based adsorbent was able to attenuate the effects of mycotoxins on the function of leukocytes and increase milk production.(AU)

Micotoxinas podem contaminar alimentos de bovinos, levando a distúrbios hepáticos, imunodepressão e redução da produção. Embora existam estudos mostrando os benefícios de adsorventes em animais de produção, há limitado conhecimento sobre sua ação quando a intoxicação de bovinos ocorre por dietas naturalmente contaminadas. O presente estudo objetivou avaliar se a administração oral diária do adsorvente de micotoxinas, argila bentonita a base de aluminossilicato durante 56 dias atenuaria alterações promovidas pelas micotoxinas em 18 vacas leiteiras, multíparas no meio do estágio da lactação, consumindo dieta contendo fumonisina B1 e B2, zearalenona (ZEA) e desoxinivalenol (DON). Os animais foram divididos, grupos tratamento (GT, n=9) e controle (GC, n=9), e submetidos às avaliações de enzimas e função hepática, hematológicas, metabolismo oxidativo de leucócitos pela técnica do nitroazul de tetrazolium (NBT) e análise físico-química do leite, a cada semana, durante dois meses, totalizando oito análises. Observou-se que o uso do adsorvente promoveu redução da excreção láctea de aflatoxina M1 (AFM1), aumento de proteína (p = 0,03) e albumina séricas (p= 0,0001), e aumento do metabolismo oxidativo leucocitário a partir do dia 24 (p = 0,05). Houve também elevação da produção leiteira a partir do 16º dia de tratamento (p= 0,008). Não foi observado melhora nos índices físico-químicos do leite dos animais e houve pouca influência na atividade das enzimas hepáticas. Conclui-se que o uso do adsorvente a base de aluminossilicato foi capaz de atenuar os efeitos das micotoxinas na função de leucócitos e de elevar a produção leiteira.(AU)

In vitro evaluation of hydroxyapatite, chitosan, and carbon nanotube composite biomaterial to support bone healing / Avaliação in vitro do biomaterial compósito à base de hidroxiapatita, quitosana e nanotubo de carbono como adjuvante no reparo ósseo

Hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were developed to improve bone healing. Previous studies suggested that a combination of biomaterials and mesenchymal stem cells (MSCs) can potentially help promote bone regeneration. In the present study, we first developed hydroxyapatite, chitosan, and carbon nanotube composite biomaterial. Then, the effect of different concentrations of the extract on the viability of Vero cells (ATCC CCL-81) and MSCs obtained from sheep bone marrow using methylthiazol tetrazolium (MTT) and propidium iodide (PI) assays were evaluated. The biomaterial group demonstrated an absence of cytotoxicity, similar to the control group. Samples with 50% and 10% biomaterial extract concentrations showed higher cell viability compared to samples from the control group (MTT assay). These results suggest that the presence of this composite biomaterial can be used with MSCs. This study also concluded that hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were not cytotoxic. Therefore, these could be used for performing in vivo tests.(AU)

O compósito à base de hidroxiapatita, quitosana e nanotubo de carbono foi desenvolvido com o intuito de auxiliar na consolidação óssea. Estudos anteriores sugerem que a combinação de substitutos ósseos e células-tronco mesenquimais (CTM) podem auxiliar a potencializar e promover a regeneração óssea. No presente estudo, o biomaterial foi desenvolvido e a viabilidade e a citotoxicidade de células Vero (ATCC CCL-81) e CTM obtidas de medula óssea provenientes de ovinos utilizando ensaios metil-tiazol-tetrazólio, MTT e iodeto de propídeo (PI) foram avaliadas em diferentes concentrações de extrato desse compósito. O compósito demonstrou ausência de citotoxicidade com comportamento semelhante ao grupo controle. Amostras com 50% e 10% de concentração de extrato do compósito mostraram resultados maiores comparados ao grupo controle (ensaio MTT). Esses resultados também sugerem que a presença do biomaterial pode ser utilizada em associação a CTM. Assim, esse estudo conclui que o compósito apresentado de hidroxiapatita, quitosana e nanotubo de cabono não foi considerado citotóxico e pode ser utilizado em teste in vivo.(AU)

In vitro evaluation of hydroxyapatite, chitosan, and carbon nanotube composite biomaterial to support bone healing / Avaliação in vitro do biomaterial compósito à base de hidroxiapatita, quitosana e nanotubo de carbono como adjuvante no reparo ósseo

Hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were developed to improve bone healing. Previous studies suggested that a combination of biomaterials and mesenchymal stem cells (MSCs) can potentially help promote bone regeneration. In the present study, we first developed hydroxyapatite, chitosan, and carbon nanotube composite biomaterial. Then, the effect of different concentrations of the extract on the viability of Vero cells (ATCC CCL-81) and MSCs obtained from sheep bone marrow using methylthiazol tetrazolium (MTT) and propidium iodide (PI) assays were evaluated. The biomaterial group demonstrated an absence of cytotoxicity, similar to the control group. Samples with 50% and 10% biomaterial extract concentrations showed higher cell viability compared to samples from the control group (MTT assay). These results suggest that the presence of this composite biomaterial can be used with MSCs. This study also concluded that hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were not cytotoxic. Therefore, these could be used for performing in vivo tests.(AU)

O compósito à base de hidroxiapatita, quitosana e nanotubo de carbono foi desenvolvido com o intuito de auxiliar na consolidação óssea. Estudos anteriores sugerem que a combinação de substitutos ósseos e células-tronco mesenquimais (CTM) podem auxiliar a potencializar e promover a regeneração óssea. No presente estudo, o biomaterial foi desenvolvido e a viabilidade e a citotoxicidade de células Vero (ATCC CCL-81) e CTM obtidas de medula óssea provenientes de ovinos utilizando ensaios metil-tiazol-tetrazólio, MTT e iodeto de propídeo (PI) foram avaliadas em diferentes concentrações de extrato desse compósito. O compósito demonstrou ausência de citotoxicidade com comportamento semelhante ao grupo controle. Amostras com 50% e 10% de concentração de extrato do compósito mostraram resultados maiores comparados ao grupo controle (ensaio MTT). Esses resultados também sugerem que a presença do biomaterial pode ser utilizada em associação a CTM. Assim, esse estudo conclui que o compósito apresentado de hidroxiapatita, quitosana e nanotubo de cabono não foi considerado citotóxico e pode ser utilizado em teste in vivo.(AU)

Burial depths favor Italian ryegrass persistence in the soil seed bank

Italian ryegrass (Lolium multiflorum Lam.) is a weed broadly found in fields cultivated with wheat and barley crops. Seed inputs into the soil before harvesting winter crops increase seed bank, ensuring survival. This study evaluated the persistence of Italian ryegrass seeds subjected to burial depths over time. Experiments were carried out in the field for two years in randomized block experimental design with four repetitions. Dormant seeds harvested from mother-plants were put in nylon bags containing soil. Previously, seed viability was evaluated in a 1 % tetrazolium solution, and 50 viable seeds by repetition were buried at 0.5, 5.0, 10, and 20 cm depths. Seed persistence was evaluated by the percentage of deteriorated and remaining seeds, non-dormant seeds, abnormal seedlings, and viable and non-viable dormant seeds at 0, 30, 60, 90, 180, 360, 540, and 720 days after burial (DAB). Seed persistence increased at 10 and 20 cm of burial depth compared to seeds in the soil layers up to 5 cm. Moreover, burial depth at 10 and 20 cm showed lower percentage of deteriorated seeds (10 % lower) compared to 0.5 cm at 360 DAB. For non-dormant seeds, a higher percentage was found at 90 DAB, regardless of seed burial depth. Dormancy breakage occurred until 180 DAB, and more rapidly at 10 and 20 cm depths. At 540 DAB, more than 95 % of seeds were unviable, demonstrating short persistence of Italian ryegrass in the soil seed bank.

Quality of forage seeds sold in the state of Mato Grosso / Qualidade de sementes forrageiras comercializadas no estado de Mato Grosso

The acquisitionof high-quality seeds is essential to successfully implant areas with forage. However, to be commercialized, they must meet the minimum quality standards established by law. Inspections are necessary to guarantee the quality and identity of the commercialized seed. Thus, the objective was to evaluate the quality of tropical forage seed species collected in 2018, in the state of Mato Grosso. The survey was based on data from officers’ samples, in commercial establishments and fine consumers, collected during inspections conducted by the Agricultural Defense Institute of the State of Mato Grosso -INDEA-MT and subsequently analyzed by the Guilherme de Abreu Lima Seed Analysis Laboratory (LASGAL). Purity analysis, determination of other seeds by number (DOSN), germination test and tetrazolium test were carried out. The study showed that 75% of the analyzed samples reached the minimum requirements for commercialization in terms of physical purity; 95% of the samples were within the standard for physiological quality determined by law; and 91.9% did not exceed the maximum limits regarding the number of undesirable seeds present in a sample. In general, 36.1% of the total samples analyzed did not conform in some way to quality standards, therefore were not appropriate for commercialization.

A aquisição de sementes de alta qualidade é primordial para o sucesso da implantação de áreas com forrageiras. Contudo, para serem comercializadas, estas devem atender aos padrões mínimos de qualidade previsto em Lei. Sendo a fiscalização necessária para garantir a qualidade e identidade das sementes comercializadas. Assim, objetivou-se avaliar a qualidade das sementes de espécies forrageiras tropicais coletadas no ano de 2018, no estado de Mato Grosso. O levantamento foi baseado emdados oriundos de amostras oficiais, coletadas em estabelecimentos comerciais e consumidores finais, pela fiscalização do Instituto de Defesa Agropecuária do estado de Mato Grosso –INDEA-MT e posteriormente analisadas pelo Laboratório de Análise de Sementes Guilherme de Abreu Lima (LASGAL). Foram realizadas análises de pureza, determinação de outras sementes por número (DOSN), teste de germinação e teste de tetrazólio. O estudo demonstrou que 75% das amostras analisadas atingiram o mínimo estabelecido para comercialização quanto a pureza física; 95% das amostras estavam dentro do padrão para qualidade fisiológica exigidos pela lei; e 91,9% não ultrapassaram os limites máximos do número de sementes indesejáveis presentes em uma amostra. No geral 36,1% do total de amostras analisadas possuía alguma inconformidade no padrão de qualidade, sendo reprovadas para comercialização.

Cytotoxicity assessment of beta-lapachone in endothelial cells

The selective activity of an antineoplastic drug is related to its ability to promote cytotoxic action on tumor cells and preserve the integrity of non-neoplastic cells. Beta-lapachone is extracted from the sawdust of Ipe wood, a thick bark tree from the Ipe wood found in the Brazilian Cerrado biome. This study aimed to evaluate the cytotoxic action of beta-lapachone in an endothelial cell line. The EA.hy926 cells were seeded in two groups, G1 and G2, cultured and exposed to beta-lapachone at concentrations of 0.0, 0.01, 0.03, 0.1, 0.3, 1 and 3 & 956;M for 24 hours. G1 remained under normal cultivation conditions and G2 was subjected to oxidative stress through an ischemia and reperfusion assay, in a deoxygenated sealed chamber. The cytotoxicity assay was performed using the tetrazolium reduction method. In G1, the cytotoxicity ranged from 0.0 to 10.0%; and in G2 between 0.0 and 6.3%. No statistically significant difference was observed between the obtained values. Moreover, we found no cytotoxic action of beta-lapachone on endothelial cells, and the results point out that the drug might have preserved the cell’s integrity against oxidative stress under the conditions of this experiment. This promising result suggests the possibility of beta-lapachone as a chemotherapy drug with selective activity.

Ethanolic extract of gabirobeira (Campomanesia adamantium) leaves reduces proliferation of osteosarcoma cells in vitro

Osteosarcoma is the most commonly diagnosed malignant bone tumor in humans, with a higher incidence in children and young people. It is highly aggressive and has a high metastatic potential. Its treatment is based on both chemotherapy and surgical intervention. However, currently used chemotherapeutic agents, such as doxorubicin, have several adverse effects on the patient. Therefore, there is a growing demand for new chemotherapeutic agents that stimulate new researches, such as those involving compounds extracted from plants, such as the gabirobeira. In this study, we aimed to evaluate the cytotoxic effects of ethanolic extract, both crude and ethyl acetate, of gabirobeira leaves on osteosarcoma cells in vitro. Cytotoxicity was evaluated using the Trypan blue exclusion method and the IC50 values were calculated using the tetrazolium reduction method. The ethanolic extract of gabirobeira leaves showed a cytotoxic effect on osteosarcoma cells in vitro. The group treated with the crude extract at 1. 0& 956;L mL-1 concentration for 48 hours showed higher cytotoxicity and the lowest IC50 value for this extract was found in the 24 to 48 hours interval. The ethanolic extract of gabirobeira leaves is cytotoxic for osteosarcoma cells.

Quality of forage seeds sold in the state of Mato Grosso / Qualidade de sementes forrageiras comercializadas no estado de Mato Grosso

The acquisitionof high-quality seeds is essential to successfully implant areas with forage. However, to be commercialized, they must meet the minimum quality standards established by law. Inspections are necessary to guarantee the quality and identity of the commercialized seed. Thus, the objective was to evaluate the quality of tropical forage seed species collected in 2018, in the state of Mato Grosso. The survey was based on data from officers samples, in commercial establishments and fine consumers, collected during inspections conducted by the Agricultural Defense Institute of the State of Mato Grosso -INDEA-MT and subsequently analyzed by the Guilherme de Abreu Lima Seed Analysis Laboratory (LASGAL). Purity analysis, determination of other seeds by number (DOSN), germination test and tetrazolium test were carried out. The study showed that 75% of the analyzed samples reached the minimum requirements for commercialization in terms of physical purity; 95% of the samples were within the standard for physiological quality determined by law; and 91.9% did not exceed the maximum limits regarding the number of undesirable seeds present in a sample. In general, 36.1% of the total samples analyzed did not conform in some way to quality standards, therefore were not appropriate for commercialization.9AU)

A aquisição de sementes de alta qualidade é primordial para o sucesso da implantação de áreas com forrageiras. Contudo, para serem comercializadas, estas devem atender aos padrões mínimos de qualidade previsto em Lei. Sendo a fiscalização necessária para garantir a qualidade e identidade das sementes comercializadas. Assim, objetivou-se avaliar a qualidade das sementes de espécies forrageiras tropicais coletadas no ano de 2018, no estado de Mato Grosso. O levantamento foi baseado emdados oriundos de amostras oficiais, coletadas em estabelecimentos comerciais e consumidores finais, pela fiscalização do Instituto de Defesa Agropecuária do estado de Mato Grosso –INDEA-MT e posteriormente analisadas pelo Laboratório de Análise de Sementes Guilherme de Abreu Lima (LASGAL). Foram realizadas análises de pureza, determinação de outras sementes por número (DOSN), teste de germinação e teste de tetrazólio. O estudo demonstrou que 75% das amostras analisadas atingiram o mínimo estabelecido para comercialização quanto a pureza física; 95% das amostras estavam dentro do padrão para qualidade fisiológica exigidos pela lei; e 91,9% não ultrapassaram os limites máximos do número de sementes indesejáveis presentes em uma amostra. No geral 36,1% do total de amostras analisadas possuía alguma inconformidade no padrão de qualidade, sendo reprovadas para comercialização.(AU)

Ethanolic extract of gabirobeira (Campomanesia adamantium) leaves reduces proliferation of osteosarcoma cells in vitro

Osteosarcoma is the most commonly diagnosed malignant bone tumor in humans, with a higher incidence in children and young people. It is highly aggressive and has a high metastatic potential. Its treatment is based on both chemotherapy and surgical intervention. However, currently used chemotherapeutic agents, such as doxorubicin, have several adverse effects on the patient. Therefore, there is a growing demand for new chemotherapeutic agents that stimulate new researches, such as those involving compounds extracted from plants, such as the gabirobeira. In this study, we aimed to evaluate the cytotoxic effects of ethanolic extract, both crude and ethyl acetate, of gabirobeira leaves on osteosarcoma cells in vitro. Cytotoxicity was evaluated using the Trypan blue exclusion method and the IC50 values were calculated using the tetrazolium reduction method. The ethanolic extract of gabirobeira leaves showed a cytotoxic effect on osteosarcoma cells in vitro. The group treated with the crude extract at 1. 0& 956;L mL-1 concentration for 48 hours showed higher cytotoxicity and the lowest IC50 value for this extract was found in the 24 to 48 hours interval. The ethanolic extract of gabirobeira leaves is cytotoxic for osteosarcoma cells.(AU)

Cytotoxicity assessment of beta-lapachone in endothelial cells

The selective activity of an antineoplastic drug is related to its ability to promote cytotoxic action on tumor cells and preserve the integrity of non-neoplastic cells. Beta-lapachone is extracted from the sawdust of Ipe wood, a thick bark tree from the Ipe wood found in the Brazilian Cerrado biome. This study aimed to evaluate the cytotoxic action of beta-lapachone in an endothelial cell line. The EA.hy926 cells were seeded in two groups, G1 and G2, cultured and exposed to beta-lapachone at concentrations of 0.0, 0.01, 0.03, 0.1, 0.3, 1 and 3 & 956;M for 24 hours. G1 remained under normal cultivation conditions and G2 was subjected to oxidative stress through an ischemia and reperfusion assay, in a deoxygenated sealed chamber. The cytotoxicity assay was performed using the tetrazolium reduction method. In G1, the cytotoxicity ranged from 0.0 to 10.0%; and in G2 between 0.0 and 6.3%. No statistically significant difference was observed between the obtained values. Moreover, we found no cytotoxic action of beta-lapachone on endothelial cells, and the results point out that the drug might have preserved the cells integrity against oxidative stress under the conditions of this experiment. This promising result suggests the possibility of beta-lapachone as a chemotherapy drug with selective activity.(AU)

Burial depths favor Italian ryegrass persistence in the soil seed bank

Italian ryegrass (Lolium multiflorum Lam.) is a weed broadly found in fields cultivated with wheat and barley crops. Seed inputs into the soil before harvesting winter crops increase seed bank, ensuring survival. This study evaluated the persistence of Italian ryegrass seeds subjected to burial depths over time. Experiments were carried out in the field for two years in randomized block experimental design with four repetitions. Dormant seeds harvested from mother-plants were put in nylon bags containing soil. Previously, seed viability was evaluated in a 1 % tetrazolium solution, and 50 viable seeds by repetition were buried at 0.5, 5.0, 10, and 20 cm depths. Seed persistence was evaluated by the percentage of deteriorated and remaining seeds, non-dormant seeds, abnormal seedlings, and viable and non-viable dormant seeds at 0, 30, 60, 90, 180, 360, 540, and 720 days after burial (DAB). Seed persistence increased at 10 and 20 cm of burial depth compared to seeds in the soil layers up to 5 cm. Moreover, burial depth at 10 and 20 cm showed lower percentage of deteriorated seeds (10 % lower) compared to 0.5 cm at 360 DAB. For non-dormant seeds, a higher percentage was found at 90 DAB, regardless of seed burial depth. Dormancy breakage occurred until 180 DAB, and more rapidly at 10 and 20 cm depths. At 540 DAB, more than 95 % of seeds were unviable, demonstrating short persistence of Italian ryegrass in the soil seed bank.(AU)