Efecto de un producto natural a base de Amaranto, Quinua y Tarwi sobre el perfil Lipídico en pacientes con obesidad y diabetes Mellitus tipo 2 / Effect of a natural product based on Amaranth, Quinoa and Tarwi on lipid profile in patients with obesity and type 2 diabetes Mellitus

INTRODUCCIÓN: la obesidad es una enfermedad crónica que conlleva a la aparición de enfermedades cardiovasculares y diabetes mellitus tipo 2 (DM2). Pacientes con sobrepeso, obesidad o DM2 presentan generalmente un perfil lipídico con niveles sanguíneos elevados de colesterol, triglicéridos, LDL (lipoproteínas de baja densidad) y VLDL-c (lipoproteínas de muy baja densidad unidas al colesterol) y niveles bajos de HDL-c (lipoproteínas de alta densidad). Pseudocereales como Amaranthus caudatus (Amaranto), Chenopodium quinoa (Quinua) y Lupinus mutabilis (Tarwi) presentes en la región Andina de Bolivia, tienen propiedades potencialmente nutracéuticas, con un alto contenido de macronutrientes y diversos fitoquímicos con actividad biológica como alcaloides de quinolizidina, saponinas, triterpenos y γ-conglutina. OBJETIVO: determinar el efecto del consumo de un producto natural elaborado a base de granos de Amaranto, Quinua y Tarwi (AQT), sobre el perfil lipídico en pacientes con obesidad y DM2. MÉTODO: estudio clínico preliminar controlado, prospectivo, doble ciego y cruzado realizado en el Área de Farmacología del Instituto de Investigaciones Fármaco Bioquímicas de la Facultad de Ciencias Farmacéuticas y Bioquímicas de la Universidad Mayor de San Andrés. RESULTADOS: el consumo de un producto elaborado a base de harinas de Amaranto, Quinua y Tarwi (AQT) durante 3 meses, promovió la disminución significativa de colesterol, triglicéridos, LDL y VLDLc en pacientes diabéticos (DM2) con sobrepeso (IMC>25), sin embargo, los niveles de HDL-c no sufrieron cambios significativos. CONCLUSIÓN: por tanto, el producto AQT tiene efectos beneficiosos sobre el perfil lipídico en pacientes con sobrepeso u obesidad y con riesgo de padecer enfermedades cardiovasculares y DM2.

INTRODUCTION: obesity is a chronic disease that leads to the onset of cardiovascular diseases and type 2 diabetes mellitus (T2DM). Overweight, obese or T2DM patients generally have a lipid profile with high blood levels of cholesterol, triglycerides, LDL (low-density lipoprotein) and VLDL-c (very low-density lipoprotein) and low levels of HDL-c (high-density lipoprotein of cholesterol). Pseudocereals such as Amaranthus caudatus (Amaranth), Chenopodium quinoa (Quinoa) and Lupinus mutabilis (Tarwi) present in the Andean region of Bolivia, have potentially nutraceutical properties, with a high content of macronutrients and various phytochemicals with biological activity such as quinolizidine alkaloids, saponins, triterpenes and γ-conglutin. OBJECTIVE: to determine the effect due to natural product intake made with Amaranth, Quinoa and Tarwi (AQT) on the lipid profile in patients with obesity and T2DM. METHOD: controlled, prospective, double blinded and crossed Preliminary Clinical Assay, performed in the Pharmacology Area of Instituto de Investigaciones Fármaco Bioquímicas from Facultad de Ciencias Farmacéuticas y Bioquímicas, Universidad Mayor de San Andrés. RESULTS: the intake of a product made from Amaranth, Quinoa and Tarwi (AQT) flour for 3 months, promoted a significant decrease in cholesterol, triglycerides, LDL and VLDL-c in overweight diabetic patients (DM2) (BMI> 25) However, HDL-c levels did not undergo significant changes. CONCLUSION: therefore, the AQT product has beneficial effects on the lipid profile in patients who are overweight or obese and at risk of cardiovascular disease and T2DM.

The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity

BACKGROUND: The 11S globulin from amaranth is the most abundant storage protein in mature seeds and is well recognized for its nutritional value. We used this globulin to engineer a new protein by adding a four valinetyrosine antihypertensive peptide at its C-terminal end to improve its functionality. The new protein was named AMR5 and expressed in the Escherichia coli BL21-CodonPlus(DE3)-RIL strain using a custom medium (F8PW) designed for this work. RESULTS: The alternative medium allowed for the production of 652 mg/L expressed protein at the flask level, mostly in an insoluble form, and this protein was subjected to in vitro refolding. The spectrometric analysis suggests that the protein adopts a ß/α structure with a small increment of α-helix conformation relative to the native amaranth 11S globulin. Thermal and urea denaturation experiments determined apparent Tm and C1/2 values of 50.4°C and 3.04 M, respectively, thus indicating that the antihypertensive peptide insertion destabilized the modified protein relative to the native one. AMR5 hydrolyzed by trypsin and chymotrypsin showed 14- and 1.3-fold stronger inhibitory activity against angiotensin I-converting enzyme (IC50 of 0.034 mg/mL) than the unmodified protein and the previously reported amaranth acidic subunit modified with antihypertensive peptides, respectively. CONCLUSION: The inserted peptide decreases the structural stability of amaranth 11S globulin and improves its antihypertensive activity.

Evaluación de los extractos de Amaranthus caudatus (Amaranto), Lupinus mutabilis (Tarwi) y Linum usitatissimum (Linaza), sobre la hiperglicemia inducida por aloxano en ratones / Evaluation of the extracts of Amaranthus caudatus (Amaranto), Lupinus mutabilis (Tarwi) and Linum usitatissimum (Linaza), on the Alloxane-induced hyperglycemia in mice

La prevalencia de diabetes mellitus se incrementa en todo el mundo alcanzando a 592 millones de diabéticos el año 2035; así mismo la OMS proyecta que las muertes por diabetes se dupliquen entre los años 2005 y 2030 (OMS, 2016). En Bolivia la medicina tradicional reporta plantas medicinales a las cuales se les ha atribuido propiedades hipoglucemiantes, sin embargo en muchos casos no existen estudios científicos que avalen dichas propiedades. Este estudio se ha realizado con la finalidad de evaluar el efecto de los granos de Amaranthus caudatus (amaranto), Linum usitatissimum (Linaza) y Lupinus mutabilis (tarwi) sobre la hiperglicemia inducida por aloxano en animales de experimentación. Se administró por vía oral a ratones con hiperglicemia (glicemia > 180,6 mg/dl) una dosis de 2000 mg/kg de peso corporal de cada extracto hidro-etanólico obtenido de los granos de A. caudatus, L. usitatissimum y L. mutabilis. Los niveles de glucosa fueron medidos antes y después de la administración de los extractos. Los extractos hidro-etanolicos disminuyeron de forma significativa (p<0.05) la glucosa plasmática a las cuatro y dos horas después de su administración. El extracto de A. caudatus disminuyo la glucosa plasmática de 380 mg/dl a 260 mg/dl, el extracto de L. mutabilis disminuyo la glucosa plasmática de 310 mg/dl a 167 mg/dl, y el extracto de L. usitatissimum disminuyo la glucosa plasmática de 210,57 mg/dl a 168,14 mg/dl. Siendo el extracto de L. mutabilis el que presento mayor actividad sobre la hiperglicemia inducida por aloxano.

The prevalence of diabetes mellitus increases worldwide reaching 592 million diabetics in 2035; Likewise, the WHO projects that deaths due to diabetes double between the years 2005 and 2030 (WHO, 2016). In Bolivia, traditional medicine reports medicinal plants to which hypoglycaemic properties have been attributed, however in many cases there are no scientific studies to support these properties. This study was carried out with the purpose of evaluating the effect of Amaranthus caudatus (amaranth), Linum usitatissimum (Linseed) and Lupinus mutabilis (tarwi) on the hyperglycemia induced by alloxan in experimental animals. Mice with hyperglycemia (glycemia>10 mmol/L) were administered a dose of 2000 mg/kg body weight orally of each hydro-ethanolic extract obtained from the grains of A. caudatus, L. usitatissimum and L. mutabilis. Glucose levels were measured before and after the administration of the extracts. The hydro-ethanol extracts significantly decreased (p <0.05) the plasma glucose at four and two hours after its administration. The extract of A. caudatus decreased the plasma glucose from 380 mg / dl to 260 mg / dl, the extract of L. mutabilis decreased the plasma glucose from 310 mg / dl to 167 mg / dl, and the extract of L. usitatissimum decreased Plasma glucose from 210.57 mg / dl to 168.14 mg / dl. The extract of L. mutabilis was the one with the highest activity on the hyperglycemia induced by alloxan.

Contenido de oligoelementos y factores antinutricionales de hojas comestibles nativas de Mesoamérica / Content of trace elements and antinutritional factors in edible leaves native to Mesoamerica

Los oligoelementos son importantes constituyentes nutricionales de las hierbas comestibles. Se colectaron 11 especies, nueve nativas (Amaranthus hybridus, Cnidoscolus aconitifolius, Crotalaria longirostrata, Dysphania ambrosioides, Lycianthes synanthera, Sechium edule, Solanum americanum, S. nigrescens, S. wendlandii) y dos introducidas (Moringa oleifera, Spinacea oleracea) en dos regiones de Guatemala. Se prepararon muestras de la hierba seca, cocida y de caldo de hierba fresca. Se cuantificaron por espectrofotometría de absorción atómica los macro (N, P, K) y oligoelementos (Ca, Mg, Na, Cu, Zn, Mn), taninos por espectrofotometría y oxalatos por permanganimetría. El contenido de oligoelementos es diverso, para Zn, la hierba control S. oleracea contienen buena cantidad (90-140 ppm); de las nativas D. ambrosioides (130-160 ppm) y A. hybridus (70-80 ppm) tienen la mayor cantidad. Respecto a Fe las hierbas control tienen buena composición (S. oleracea, 220-280 ppm y M. oleifera, 105-135 ppm); de las nativas A. hybridus (90-240 ppm), C. aconitifolius (75-185 ppm) y L. synanthera (75-140 ppm) tienen buenas concentraciones. Se encontraron niveles elevados de oxalatos en S. oleracea (67.30 (5.51) mg/g), L. synanthera (56.30 (9.67) mg/g) y S. nigrescens (33.6 (5.48) mg/g); en las demás hierbas se encontraron niveles menores. Los niveles de taninos fueron bajos (0.1-0.8 mg/g) para todas las especies. Se demuestra que cuatro especies nativas tienen un buen contenido de oligoelementos y presentan valores menores de antinutricionales que los controles.

Trace elements are important nutritional constituents from edible herbs. Eleven species were collected in two regions of Guatemala, nine native (Amaranthus hybridus, Cnidoscolus aconitifolius, Crotalaria longirostrata, Dysphania ambrosioides, Lycianthes synanthera, Sechium edule, Solanum americanum, S. nigrescens, S. wendlandii) and two introduced (Moringa oleifera, Spinacea oleracea). Dry, cooked and broth samples were prepared. By atomic absorption spectrometry, macro (N, P, K) and trace elements (Ca, Mg, Na, Cu, Zn, Mn) were quantified, tannins by spectrophotometry, and oxalates by permanganometry. Trace elements content is diverse, for Zn, control herb S. oleracea contained high quantity (90-140 ppm); from the natives D. ambrosioides (130-160 ppm) and A. hybridus (70-80 ppm) contained high amounts. For Fe, control herbs had high composition (S. oleracea, 220-280 ppm, M. oleifera, 105-135 ppm); from the natives A. hybridus (90-240 ppm), C. aconitifolius (75-185 ppm) and L. synanthera (75-140 ppm) had the highest amounts. High levels of oxalates were demonstrated in S. oleracea (67.30 (5.48) mg/g), L. synanthera (56.30 (9.67) mg/g), and S. nigrescens (33.6 (5.48) mg/g); from the others levels.

Improvement of nutritional quality of local dishes and their acceptance by children of different ages / Melhoria de pratos típicos: qualidade nutricional e sua aceitação por crianças de várias idades

ABSTRACT Objective Food insecurity and child malnutrition are central topics in many food programs around the world; however, these indices do not diminish. The creation of strategies is necessary to increase the effectiveness of nutritional improvement programs. The objective of this research was to improve the nutritional quality of three local dishes included in the regular diet of the population by adding amaranth and to evaluate their acceptance by children from three communities in Tochimilco, Puebla, Mexico. Methods The study was carried out in three communities of "Tochimilco", Puebla, Mexico, in order to evaluate three local dishes: banana smoothie, beans and corn tortillas, which were chosen by mothers of the region in participative workshops on preparation of local dishes enriched with amaranth. Two sensory tests were applied: paired-comparison and hedonic scale to 208 and 210 children of preschool and scholar age, respectively. Results It was found that consumers showed similar preference for banana smoothie and beans enriched with amaranth compared to non-enriched dishes; while the amaranth enriched tortilla gained more preference in the different statistical tests and it was the dish with higher content of amino acids. Sensory attributes were similarly assessed and there were no significant differences among rural and urban communities and school year. Conclusion Children easily accept the addition of amaranth to the dishes they are accustomed to, as it does not change their food culture. It is possible to improve child nutrition by designing healthier dishes and incorporating them into the daily cooking of those who prepare the food.

RESUMO Objetivo A insegurança alimentar e a desnutrição infantil são temas centrais de muitos programas alimentares ao redor do mundo; no entanto, esses índices não diminuem. É preciso criar estratégias para aumentar a eficácia dos programas de desenvolvimento nutricional. A presente pesquisa teve como objetivo melhorar a qualidade nutricional de três pratos típicos da dieta normal da população local, adicionando amaranto às comidas típicas e avaliando a aceitação dele por crianças de três comunidades em Tochimilco, Puebla, México. Métodos O estudo foi realizado nas três comunidades para avaliar os três pratos típicos desta mesma região: vitamina de banana, feijão e as tortilhas de milho. Estas comidas foram escolhidas por mães da região em oficinas participativas de preparação de pratos locais enriquecidos com amaranto. Dois testes sensoriais foram aplicados: comparação pareada e escala hedônica para 208 e 210 crianças em idade pré-escolar e escolar, respectivamente. Resultados Verificou-se que os consumidores mostraram uma preferência semelhante pela vitamina de banana e feijão enriquecido com amaranto do que os mesmos não enriquecidos; enquanto a tortilha de milho enriquecida com amaranto ganhou mais preferência nos diferentes testes estatísticos e foi o prato com maior teor de aminoácidos. Os atributos sensoriais foram avaliados da mesma forma e não houve diferenças significativas entre as comunidades rurais e urbanas e as notas escolares. Conclusão As crianças aceitam facilmente a incorporação de amaranto nos pratos típicos da dieta delas, pois não repre-sentou nenhuma alteração da sua cultura alimentar. É possível melhorar a nutrição infantil, criando pratos mais saudáveis e incorporando-os na culinária cotidiana daqueles que preparam a comida.

Impact of the incorporation of probiotic strains and fruit by-products in fermented synbiotic soy product and on the composition and metabolic activity of the gut microbiota in vitro / Impacto da incorporação de cepas probióticas e de subprodutos de frutas em um produto fermentado de soja simbiótico e sobre a composição e a atividade metabólica da microbiota intestinal humana in vitro

The present study aimed to develop a fermented soy beverage containing fruit by-products and probiotics and to evaluate the impact of this product on the composition and metabolic activity of the human intestinal microbiota using an in vitro simulation model of the intestinal conditions (TIM-2). Therefore, the present study was divided into three stages. Stage I was based on obtaining, processing and physical-chemical, microbiological and functional characterization of fruit by-products (acerola, orange, mango, and passion fruit) and soybean (okara), as well as amaranth flour. Additionally, the ability to use these vegetable by-products and amaranth flour by probiotic and non-probiotic strains was evaluated. The results showed that the acerola byproduct presented the highest dietary fibre content (48.46 g/100 g) among the by-products tested, as well as amaranth flour. Orange and passion fruit by-products were the substrates that most promoted the growth of bacterial populations, including strains of Escherichia coli and Clostridium perfringens. On the other hand, the acerola by-product was the substrate that showed the highest selectivity for beneficial bacteria. Also, in this stage, ten probiotic strains (seven lactobacilli and three bifidobacteria) and three starter strains (Streptococcus thermophilus) were tested for their ability to deconjugate bile salts and for proteolytic activity against milk and soy proteins. The results showed that none of the tested strain showed proteolytic ability against milk and soybean proteins. In addition, the probiotic strains Lactobacillus acidophilus LA-5 and Bifidobacterium longum BB-46 deconjugated more types of bile acids tested, and the strains of S. thermophilus tested showed no ability to deconjugate bile salts. Next, the acerola by-product (ABP) and the probiotic strains LA-5 and BB-46 were selected to continue stage II of the study (development of a fermented soy beverage). For this purpose, a 23 factorial design was used, in a total of 8 trials with three replicates of each one, and the effects of the probiotic strains and the acerola by-product on the physical-chemical, microbiological, and sensory characteristics of these fermented soy beverages were evaluated. At the same time, probiotic viability and survival under in vitro gastrointestinal (GI) simulated conditions were evaluated in fermented soy beverage (FSB). The results showed that the presence of BB-46 and ABP affected the sensory acceptability of FSB negatively. ABP also led to significant differences in the texture profile of the FSB (P<0.05). Populations of probiotic strains ranged from 7.0 to 8.2 log CFU equivalent/mL during 28 days of refrigerated storage (4° C) of FBS, and the co-culture (LA-5+BB-46) and the ABP did not affect the viability of both microorganisms significantly (P> 0.05). However, ABP increased the survival of BB-46 under in vitro simulated GI conditions significantly. For stage III, a 22 experimental design was performed. To evaluate the impact of these FBS on the composition and metabolic activity of the intestinal microbiota of lean and obese humans, a validated in vitro model called TIM-2 was used, available at the Maastricht University (Venlo, The Netherlands), which simulates normal conditions of the lumen of the proximal colon, with all parameters controlled by a computer. Samples were collected from TIM-2 to quantify probiotic microorganisms (LA-5 and BB-46), Lactobacillus spp., Bifidobacterium spp., and total bacteria, using the quantitative PCR method (qPCR) and the intestinal microbiota profile was determined using an Illumina Mysec Next Generation Sequencing (NGS) method. Concentrations of shortchain fatty acids and branched-chain fatty acids and lactate produced by the different microbiotas during fermentation in TIM-2 were also determined. The results showed that the lean microbiota presented the high production of acetate and lactate than the microbiota of obese individuals. Significant reductions in Bifidobacterium populations in the lean microbiota were observed at 0 and 48 h of an assay for all experimental meals, except for the meal that had the probiotic combination (LA-5 and BB-46) and the ABP supplementation, which showed an increased total Bifidobacterium and Lactobacillus populations throughout the experimental period for both microbiotas tested. The FSB supplemented with ABP presented the best characteristics regarding the modulation of the obese microbiota, with an increase in Bifidobacterium spp. and Lactobacillus spp. Additionally, after 48 hours of intervention in TIM-2, the obese microbiota was apparently similar to the lean microbiota, showing a beneficial modulation of this microbiota. The results suggest that the fermented soy beverage supplemented with the acerola by-product and the probiotic strains may present beneficial health effects. However, clinical studies are required to complement and confirm the results observed in the in vitro assays

O presente trabalho visou desenvolver uma bebida fermentada de soja adicionada de resíduos de frutas e suplementada com cepas probióticas e avaliar o impacto desse produto sobre a composição e a atividade metabólica da microbiota intestinal humana, utilizando um modelo de simulação in vitro das condições intestinais (TIM-2). Para tanto, o presente trabalho foi dividido em três etapas. A etapa I foi baseada na obtenção, processamento e caracterização físico-química, microbiológica e funcional de subprodutos de frutas (acerola, laranja, manga e maracujá) e soja (okara), bem como da farinha de amaranto. Adicionalmente, a capacidade de utilização desses subprodutos vegetais e da farinha de amaranto por cepas probióticas e não probióticas foi avaliada. Os resultados mostraram que o subproduto de acerola apresentou o maior conteúdo de fibras alimentares totais (48,46 g/100 g) dentre os subprodutos testados, bem como a farinha de amaranto. Os subprodutos de laranja e maracujá foram os substratos que mais promoveram a multiplicação das populações bacterianas, incluindo das cepas de Escherichia coli e Clostridium perfringens. Por outro lado, o subproduto de acerola foi o substrato que apresentou a maior seletividade para bactérias benéficas. Ainda nessa etapa, dez cepas probióticas (sete lactobacilos e três bifidobacterias) e três cepas starter (Streptococcus thermophilus) foram testadas quanto à sua capacidade de desconjugação de sais biliares e atividade proteolítica frente às proteínas do leite e da soja. Os resultados revelaram que nenhuma cepa testada apresentou capacidade de proteólise das proteínas do leite e da soja. Adicionalmente, as cepas probióticas Lactobacillus acidophilus LA-5 e Bifidobacterium longum BB-46 desconjugaram a maior quantidade de ácidos biliares testados e as cepas de S. thermophilus testadas não apresentaram capacidade de desconjugação de sais biliares. Após a análise dos resultados da etapa I, o resíduo de acerola (ABP) e as cepas probióticas LA-5 e BB-46 foram selecionadas para dar continuidade à etapa II do estudo(desenvolvimento de uma bebida fermentada a base de soja). Para esse fim, foi utilizado um delineamento experimental do tipo fatorial 23, totalizando 8 ensaios com três repetições de cada, e foram avaliados os efeitos das cepas probióticas e do subproduto de acerola sobre as características físico-químicas, microbiológicas e sensoriais dessas bebidas fermentadas de soja. Paralelamente, foram realizadas análises da sobrevivência das cepas probióticas frente às condições gastrintestinais simuladas in vitro nas bebidas fermentadas de soja (FSB). Os resultados mostraram que a presença de BB-46 e ABP afetaram negativamente a aceitabilidade sensorial das FSB. O ABP também levou a diferenças significativas no perfil de textura das FSB (P<0,05). As populações das cepas probióticas nas diferentes formulações de FSB variaram de 7,0 a 8,2 log de UFC equivalente/mL durante os 28 dias de armazenamento (4 ºC) e a co-cultura (LA-5+BB-46) e o ABP não afetaram (P>0,05) a viabilidade de ambos os microrganismos. No entanto, ABP aumentou significativamente a sobrevivência de BB-46 frente às condições gastrintestinais sumuladas in vitro. Para a etapa III do presente estudo, um delineamento experimental fatorial 22 foi realizado. Para a avaliação do impacto dessas FSB sobre a composição e atividade metabólica da microbiota intestinal de humanos eutróficos e obesos, foi utilizado um modelo in vitro TIM-2 na Maastricht University (Venlo, Holanda), que simula as condições normais do lúmen do cólon proximal, com todos os parâmetros controlados por um computador. Amostras foram coletadas do TIM-2 para a quantificação dos microrganismos probióticos (LA-5 e BB-46), Lactobacillus spp., Bifidobacterium spp. e bactérias totais, utilizando o método de PCR quantitativo (qPCR), e o perfil da microbiota intestinal foi determinado utilizando Next-Generation Sequencing (NGS) Illumina Mysec. A concentração de ácidos graxos de cadeia curta e de cadeia ramificada e lactato produzidos pelas diferentes microbiotas durante a fermentação no TIM-2 também foi determinada. Os resultados mostraram que a microbiota de humanos eutróficos apresentou uma alta produção de acetato e lactato em comparação com a microbiota de obesos. Reduções significativas das populações de Bifidobacterium na microbiota de eutróficos foram observadas entre 0 e 48 h de ensaio para todas as refeições experimentais, exceto para a refeição que apresentou a combinação probiótica (LA-5 e BB-46) e a suplementação com ABP, que apresentou aumento de Bifidobacterium e Lactobacillus totais durante todo o período de análise para ambas as microbiotas testadas. As FSB suplementadas com ABP apresentaram os melhores resultados em relação à modulação da microbiota de humanos obesos, com o aumento Bifidobacterium spp. e Lactobacillus spp. Adicionalmente, após 48 horas de intervenção no TIM-2, a microbiota de obesos foi aparentemente similar à microbiota de eutróficos, mostrando uma modulação benéfica dessa microbiota. Os resultados sugerem que as bebidas fermentadas de soja suplementadas com o subproduto de acerola e cepas probióticas podem apresentar efeitos benéficos à saúde. No entanto, estudos clínicos são necessários para complementar e confirmar os resultados observados nos ensaios in vitro

Efeito de peptídeos do grão de amaranto (Amaranthus cruentus L. ) sobre os mecanismos de absorção e síntese de colesterol / Effect of amaranth grain peptides (Amaranthus cruentus L. ) on the mechanisms of absorption and synthesis of cholesterol

Introdução: Doenças cardiovasculares constituem importante causa de morte em todo mundo e a hipercolesterolemia está diretamente relacionada como fator agravante desta morbidade. A dieta desempenha papel importante neste processo e alguns alimentos como o amaranto, especialmente sua proteína, tem mostrado capacidade de redução do colesterol plasmático. Estudos sugerem que este efeito está relacionado a peptídeos formados durante a digestão da sua proteína, os quais desempenham um papel importante na regulação e modulação do metabolismo lipídico. Os efeitos hipocolesterolêmicos, já observados, indicam o uso da proteína do amaranto como um composto bioativo direcionado para a promoção da saúde. Considerando que os efeitos hipocolesterolêmicos destes peptídeos são complexos e há diversas hipóteses formuladas, torna-se importante a realização de estudos visando avaliar a interação dos peptídeos na absorção intestinal do colesterol e da sua modulação genética. Objetivos: Verificar os efeitos do hidrolisado da farinha do grão de amaranto na absorção de colesterol e modulação de genes ABCA1, ABCG1, NPC1L1, AMPK, HMGR e SREBP-2em células Caco-2, e modulação dos genes ABCG8, HMGR, SREBP-2 e AMPKem enterócitos de hamsters. Metodologia: O amaranto foi triturado, sua farinha desengordurada e sua proteína isolada, com posterior digestão in vitro e filtração dos peptídeos. Três experimentos in vitro foram conduzidos com as células: permeação de hidrolisado, permeação de colesterol e de efeito sob a expressão gênica. No primeiro, o hidrolisado proteico de amaranto foi permeado em culturas celulares de Caco-2 no tempo de 2 horas. O permeato foi coletado e analisado por LC/MS/MS. No segundo, o hidrolisado de amaranto foi incorporado a micelas de colesterol e incubados em culturas celulares, nas concentrações de 1,0 mg/ml, e 3,0 mg/ml em tempos de 2h. Também em concentrações de 3,0 mg/ml foi adicionado albumina e caseína para efeito comparativo. O conteúdo de colesterol na porção apical e basolateral foi analisado em HPLC. O terceiro experimento foi avaliaçãoda exposição do hidrolisado, em concentrações de 0,5 mg/ml, 1,0 mg/ml e 3,0 mg/ml, em tempos de 2h e 12h. Após este período, foi realizada a extração de RNA total, avaliação de rendimento e integridade do material; medida quantitativa de expressão de RNAm por RT-PCR e quantificação relativa da expressão por ?CT dos genes ABCA1, ABCG1, ABCG8, NPC1L1, AMP1, HMGR e SREBP-2das células Caco-2 e tecido intestinal de hamsters, coletados em ensaios anteriores. Resultados: Na permeação de colesterol não houve diferença entre as concentrações dos hidrolisados proteicos e controle, porém o hidrolisado de amaranto em 1,0 mg/ml demonstrou uma tendência em diminuir a absorção de colesterol (p = 0,05). Na exposição das células Caco-2 aos peptídeos por 2h, houve uma diminuição nas concentrações de RNAm dos genes ABCA1, NPC1L1, AMPK, HMGR e SREBP-2 nas concentrações de 3,0 mg/ml. O tempo de exposição de 12h apresentou resultados semelhantes ao tempo de 2h. Somente a expressão gênica de ABCG8foi influenciada pelo isolado proteico de amaranto no experimento in vivo. Conclusão: A partir do exposto, podemos concluir que os peptídeos do grão de amaranto influenciam o metabolismo de colesterol por mecanismos genéticos. Portanto, torna-se uma alternativa a ser introduzida na dieta de indivíduos saudáveis e em pacientes com hipercolesterolemia, visando a prevenção de agravos e como estratégia de terapia adicional no controle dos níveis de LDL-c plasmático. Contudo, mais experimentos in vivo e em humanos são necessários para estabelecer a dose efetiva para consumo

Introduction: Cardiovascular diseases are an important cause of death worldwide and hypercholesterolemia is directly related as an aggravating factor of this morbidity. Diet plays an important role in this process and some foods such as amaranth, especially its protein, have shown ability to lower plasma cholesterol. Studies suggest that this effect is related to peptides formed during the digestion of their protein, which play an important role in the regulation and modulation of lipid metabolism. The hypocholesterolemic effects, already observed, indicate the use of amaranth protein as a bioactive compound aimed to promoting health. Considering that the hypocholesterolemic effects of these peptides are complex and there are several hypotheses formulated, it is important to carry out studies to evaluate the interaction of peptides in the intestinal absorption of cholesterol and its genetic modulation. Objectives: To verify the effects of amaranth grain flour hydrolyzate on cholesterol uptake and ABCA1, ABCG1, NPC1L1, AMPK, HMGR and SREBP-2 genes modulation in Caco-2 intestinal cells, and modulation of ABCG8, HMGR, SREBP-2 genes and AMPK in hamster intestinal cells. Methodology: Amaranth was crushed, the created flour was defatted and its protein isolated, with subsequent in vitro digestion and filtration of the peptides. Three in vitro experiments were conducted with the cells: hydrolyzate permeation, cholesterol permeation and genetic expression. In the first, the amaranth protein hydrolyzate was permeated in Caco-2 cell cultures in the time of 2 hours. The permeate was collected and analyzed by LC/MS/MS. In the second, the amaranth hydrolyzate was incorporated into cholesterol micelles and incubated in cell cultures at concentrations of 1.0 mg/ml and 3.0 mg/ml in times of 2 h. Also, at concentrations of 3.0 mg/ml albumin and casein were added for comparison. Cholesterol content in the apical and basolateral portion was analyzed by HPLC. The third experiment was to evaluate the exposure of the hydrolyzate at concentrations of 0.5 mg/ml, 1.0 mg/ml and 3.0 mg/ml, in times of 2 h and 12 h. After this period, the extraction of total RNA, evaluation of yield and integrity of the material was performed; quantitative measurement of mRNA expression by RT-PCR and relative quantification of ?CT expression of the ABCA1, ABCG1, ABCG8, NPC111, AMPK, HMGR and SREBP-2 genes from Caco-2 cells and hamster intestinal tissue, collected in previous assays, were finalized. Results: In cholesterol permeation there was no difference between the concentrations of the protein hydrolysates and control, but the amaranth hydrolyzate at 1.0 mg/ml showed a tendency to decrease the cholesterol absorption (p = 0.05). Exposure of Caco-2 cells to peptides for 2 h resulted in a decrease in ABCA1, NPC111, AMPK, HMGR and SREBP-2 mRNA levels at concentrations of 3.0 mg/ml. The exposure time of 12h presented results similar to the time of 2h. Only the gene expression of ABCG8 was influenced by the amaranth protein isolate in the in vivo experiment. Conclusion: From the above, we can conclude that amaranth peptides influence the metabolism of cholesterol by genetic mechanisms. Therefore, it becomes an alternative to be introduced in the diet of healthy individuals and in patients with hypercholesterolemia, aiming at the prevention of aggravations and as a strategy of additional therapy in the control of plasma LDL-c levels. However, more studies should bedone with animals and humans to define the dose-efficiency for diet.

N deposition affects allelopathic potential of Amaranthus retroflexus with different distribution regions

ABSTRACT This study aims to determine the allelopathic potential of Amaranthus retroflexus (Ar) with different climatic zones on seed germination and growth of A. tricolor (At) treated with a gradient N addition. Ar leaf extracts only displayed significantly allelopathic potential on the underground growth of Ar but not the aboveground growth of At. The allelopathic potential of Ar leaf extracts on root length of At were enhanced under N addition and there may be a N-concentration-dependent relationship. The effects of the extracts of Ar leaves that collected from Zhenjiang on seed germination and growth of At may be higher than that collected from Jinan especially on root length of At under medium N addition. This reason may be the contained higher concentration of secondary metabolites for the leaves of plants that growths in high latitudes compare with that growth in low latitudes. This phenomenon may also partly be attributed to the fact that Ar originated in America and/or south-eastern Asia which have higher similarity climate conditions as Zhenjiang rather than Jinan. The allelopathic potential of Ar on seed germination and growth of acceptor species may play an important role in its successful invasion especially in the distribution region with low latitudes.

Evaluación del contenido de selenio en hojas de hierbas nativas de uso tradicional en la alimentación del guatemalteco / Evaluation of selenium content in leaves of native herbs of traditional food use in Guatemala

El selenio (Se) es un elemento esencial para el ser humano que se encuentra en pequeñas cantidades en los suelos, pero se acumula en ciertas plantas, proporcionando beneficios como antioxidante, antiinflamatorio y quemopreventivo por la presencia de unas 25 selenoproteínas que participan en diversas acciones de bienestar, lactancia, desarrollo, reproducción y salud de la progenie. El objetivo de este estudio fue evaluar el contenido de Se en hojas de vegetales utilizados tradicionalmente en la alimentación guatemalteca. Se colectaron hojas de materiales cultivados para los mercados locales de nueve hierbas nativas (Amaranthus hybridus, Cnidoscolus aconitifolius, Crotalaria longirostrata, Dysphania ambrosioides, Lycianthes synanthera, Sechium edule, Solanum americanum, Solanum nigrescens y Solanum wendlandii) y dos introducidas de reconocido uso alimenticio (Moringa oleifera y Spinacia oleracea), se secaron en un horno de convección forzada para lograr una humedad < 10% y se digirieron 0.25 ± 0.02 g de hojas en una mezcla de ácido nítrico y ácido perclórico que se calentó hasta la digestión total de la materia. El Se fue determinado por el método de reflexión total de rayos X, utilizando un estándar interno de itrio (Y) el que se midió utilizando reflectores de cuarzo en un espectrómetro de reflexión total de rayos X. De todas las especies evaluadas, únicamente A. hybridus demostró cantidades cuantificables de Se. Se determinó que 100 g de materia vegetal seca de A. hybridus proporciona 0.355 mg de Se, por lo que su consumo semanal puede contribuir con el requerimiento de este micronutriente para un adulto.

Selenium (Se) is an essential element for the human being; it is in small amounts in the soil but it accumulates in certain plants, providing benefits as antioxidant, anti-inflammatory and chemopreventive, due to the presence of about 25 selenoproteins that participate in different welfare and development actions, lactation, reproduction and health of the progeny. This study aimed to assess Se content in leaves of nine native plants traditionally used in Guatemalan food (Amaranthus hybridus, Cnidoscolus aconitifolius, Crotalaria longirostrata, Dysphania ambrosioides, Lycianthes synanthera, Sechium edule, Solanum americanum, Solanum nigrescens and Solanum wendlandii) and two internationally uses herbs (Moringa oleifera, Spinacia oleracea). Se was determined by total reflection X-ray method. Plants were dried in a forced convection oven to constant weight, then were digested by weighing 0.25 ± 0.02 g of dry plant material with a mixture of nitric and perchloric acid, and warmed to achieve complete digestion. Using a yttrium (Y) internal standard were measured using quartz reflectors Spectrometer Total reflection X-ray. Of all native plant species tested, only A. hybridus there were measurable amounts of Se. It was determined that 100 g of dry plant material of A. hybridus provides 0.355 mg of Se, so its weekly consumption by an adult might contribute to satisfied the requirement of this microelement.

Parámetros de calidad de harinas de Amaranthus caudatus Linnaeus (amaranto), Chenopodium quinoa Willd (quinua), Chenopodium pallidicaule Aellen (kañahua), Lupinus mutabilis Sweet (tarwi) / Quality parameters of flour of Amaranthus caudatus Linnaeus (amaranth), Chenopodium quinoa Willd (quinoa), Chenopodium pallidicaule Aellen (kañahua), Lupinus mutabilis Sweet (tarwi)

Las especies vegetales Amaranthus caudatus Linnaeus (amaranto), Chenopodium quinoa Willd (quinua), Lupinus mutabilis Sweet (tarwi), Chenopodium pallidicaule Aellen (kañahua), llamados también granos andinos, son desde hace mucho tiempo la base de alimentación de muchas familias en nuestro país, estos granos son reconocidos por su alto valor nutricional. En la actualidad varias empresas de nuestra región comercializan estos granos en su forma procesada. Este trabajo tuvo como finalidad establecer los parámetros de calidad e identificación de las harinas de amaranto, quinua, kañahua y tarwi provenientes de los municipios de Ancoraimes, Tomina, Huancané y Peñas, para ello se realizó el análisis micrográfico encontrándose almidón, aleurona y grasa como principales elementos. El análisis fisicoquímico realizado reportó un contenido de humedad en quinua de 6,03%, cenizas totales 2,52% y 3,8 ml como índice de hinchamiento. En amaranto un contenido de humedad de 5,76%, cenizas totales 2,86% y 6,8 ml como índice de hinchamiento. Tarwi reportó un contenido de humedad de 6,69%, cenizas totales 3,53%, y 3,6 ml como de índice de hinchamiento. Kañahua reportó un contenido de humedad de 5,82%, cenizas totales 3,53% y 4,75ml como índice de hinchamiento. El análisis químico cualitativo en los granos muestra la presencia mayoritaria de flavonoides, aminoácidos, antocianidinas, taninos.

The plant species Amaranthus caudatus Linnaeus (amaranth), Chenopodium quinoa Willd (quinoa), Lupinus mutabilis Sweet (tarwi), Chenopodium pallidicaule Aellen (kañahua), also called Andean grains, have long been the food base of many families in our Country, these grains are recognized for their high nutritional value. At present several companies of our region commercialize these grains in its processed form. The aim of this work was to establish the parameters of quality and identification of the amaranth, quinoa, kañahua and tarwi flours from the municipalities of Ancoraimes, Tomina, Huancané and Peñas. For this purpose, the micrographic analysis was performed with starch, aleurone and fat as main elements. The physicochemical analysis carried out reported a moisture content in quinoa of 6.03%, total ash 2.52% and 3.8 ml as swelling index. In amaranth a moisture content of 5.76%, total ash 2.86% and 6.8 ml as swelling index. Tarwi reported a moisture content of 6.69%, total ash 3.53%, and 3.6 ml as the index of swelling. Kañahua reported a moisture content of 5.82%, total ash 3.53% and 4.75ml as swelling index. The qualitative chemical analysis in the grains shows the majority presence of flavonoids, amino acids, anthocyanidins, tannins.

Utilização de ingredientes sucedâneos ao trigo na elaboração de bolos sem glúten / Substitutes ingredients of wheat for making gluten-free cakes

Apesar do aumento da oferta de produtos sem glúten, observa-se ainda a limitação de opções no mercado de panificados. O objetivo deste estudo foi determinar as características físico-químicas e sensoriais de bolos formulados com farinhas substitutivas ao trigo. Duas formulações contendo farinhas mistas substitutivas ao trigo (amaranto, quinoa, soja e fécula de mandioca) e uma controle (contendo trigo) foram analisadas quanto às medições e composição centesimal. Coliformes totais, pH e acidez total titulável (ATT) foram determinados no 1° e 4° dias de armazenamento. Aceitação e preferência foram avaliadas por meio de testes sensoriais. Os dados foram submetidos à análise de variância, seguida de testes paramétricos e não-paramétricos a 5% de probabilidade. Os bolos apresentaram características físicas de medições similares à formulação controle. A formulação com maior teor de soja apresentou maior teor de lipídios quando comparada com o controle. Os bolos com farinhas mistas obtiveram aceitação e preferência semelhantes entre si, porém inferiores à amostra controle. As amostras mantiveram-se estáveis quanto ao pH e ATT durante os quatro dias de armazenamento, não havendo crescimento de coliformes totais. Conclui-se que há viabilidade na formulação destes produtos com boa aceitação e valor nutricional agregado.

Despite the increase in the supply of gluten-free products, an option limitation in the bread-making market is still observed. The objective of this study was to determine the physicochemical and sensorial characteristics of the cakes formulated with the wheat substitute flours. Two formulations containing mixed wheat substitutes (amaranth, quinoa, soybean and cassava starch) and a control sample (containing wheat) were analyzed on the measurements and proximate composition. Total coliforms, pH and titratable total acidity (TTA) were determined at the 1st and 4th days of storage. Acceptance and preference were assessed by means of sensory tests. The data were evaluated by the variance analysis,followed by parametric and non-parametric tests at 5% probability. The cakes presented physical characteristics of measurements similar to the control formulation. The formulation containing the highest soybean contents showed the major lipid contents when compared to the control. The mixed-flour cakes had similar acceptance and preference, but lower than the control sample. The samples pH and TTA were stable during the four days of storage, and no growth of total coliforms occurred. Therefore, it is viable the formulation of these products with good acceptance and nutritional value.

Avaliação de biodisponibilidade e mecanismos de ação hipocolesterolemizante de peptídeos do amaranto (Amaranthus cruentus L. BRS-Alegria) / Evaluation of bioavailability and hypocholesterolemic mechanisms of peptides from Amaranth (Amaranthus cruentus L. BRS-Alegria)

Introdução: Doenças cardiovasculares constituem importante causa de morte em todo mundo e a hipercolesterolemia está diretamente relacionada a este problema de saúde pública. A dieta desempenha papel importante neste processo e alguns alimentos, como o amaranto (Amaranthus cruentus L. BRSAlegria), têm mostrado capacidade de redução do colesterol plasmático. Estudos sugerem que este efeito está relacionado a peptídeos liberados durante a digestão das proteínas, os quais atuam na modulação do metabolismo lipídico. Considerando-se que os efeitos da digestão gastrointestinal e da absorção destes peptídeos são claramente complexos torna-se importante a realização de estudos visando avaliar bioacessibilidade e mecanismos de ação destes peptídeos nos locais alvo do organismo. Objetivo: Analisar a biodisponibilidade de peptídeos em modelos animais após ingestão de isolado proteico de amaranto e relacioná-la com parâmetros ligados ao metabolismo do colesterol. Métodos: O amaranto teve sua proteína isolada. Os peptídeos da proteína do amaranto foram analisados após digestão in vitro. Dois experimentos in vivo foram conduzidos: um de fase aguda e outro de média duração. No primeiro, o isolado proteico de amaranto foi administrado a ratos e os peptídeos no sangue foram monitorados por 2 horas para verificação de fragmentos que resistissem à digestão gastrointestinal. O experimento in vivo 2 consistiu na alimentação de 3 grupos de hamster, um com dieta recomendada pela AIN93 (grupo N) e dois com dietas hipercolesterolêmicas por 21 dias, contendo a proteína de amaranto como única proteína da ração (grupo I), comparada ao controle de caseína (grupo H). Neste experimento foram analisados no plasma: peptídeos, colesterol total e frações; nas fezes: colesterol total e ácidos biliares; no fígado: colesterol, lipídeos totais, ácidos graxos, atividade enzimática da Hmgcr, expressão de Hmgcr, Srebf2, Lxr, Abca1, Abcg8 e Ampk. Resultados e discussão: Foram identificados fragmentos peptídicos provenientes da digestão in vitro do isolado proteico de amaranto, e outras dezenas de sequencias peptídicas em ratos após administração aguda de amaranto foram analisadas. Destaca-se a identificação do peptídeo ALGV, presente em proteína do amaranto de acordo com banco de dados, e similar a fragmentos com ação hipocolesterolemizante. No sangue de hamsters foram encontrados seis peptídeos com 100 por cento de cobertura e similaridade a base de dados de proteínas de amaranto, merecendo investigação sobre seus efeitos. Verificou-se que o isolado proteico de amaranto foi capaz de suprimir a hipercolesterolemia quando a dieta hipercolesterolemizante foi introduzida em paralelo a este ingrediente, com valores inferiores em 72 por cento (triglicerídeos), 64 por cento (colesterol total), 80 por cento (LDL-c) do grupo I em relação ao grupo H. Foi observada ainda menor concentração de colesterol e lipídeos totais no fígado dos animais do grupo I em relação ao grupo H (177 x 464 mg de colesterol/100 g de tecido; 2,06 x 2,86 g de lipídeos/100 g de tecido, respectivamente). Parâmetros lipídicos do sangue, das fezes e do fígado foram similares aos do grupo N, cuja dieta seguiu a preconização para roedores. Foi observada maior excreção de colesterol total no grupo I em relação ao grupo H, mas não houve maior excreção de ácidos biliares nas fezes. Não houve mudança na expressão dos genes analisados neste estudo, mas o amaranto reduziu a atividade da enzima Hmgcr. Postulase que parâmetros como expressão de Ldlr e atividade da Acat sejam alterados pela ingestão de amaranto. O perfil de ácidos graxos também foi modificado de forma a se assimilar ao grupo N, porém deve-se verificar parâmetros inflamatórios devido à maior proporção de ácido araquidônico em relação aos demais grupos estudados. Conclusão: Verifica-se biodisponibilidade dos peptídeos do amaranto e ação hipocolesterolemizante e hipolipemiante em diversas vias metabólicas, promovendo proteção cardiovascular

Introduction: Cardiovascular diseases are important causes of death worldwide, and hypercholesterolemia is directly related to this public health problem. Diet plays an important role in this process and some foods such as amaranth (Amaranthus cruentus L. BRS-Alegria) have been shown to reduce plasma cholesterol. Studies suggest that this effect is related to peptides released during the digestion of proteins, which would play an important role in the modulation of lipid metabolism. Considering that the effects of gastrointestinal digestion and the absorption of these peptides are clearly complex, it is important to carry out studies aiming to evaluate their bioaccessibility and evaluation of the mechanisms of action of these peptides in the target sites of the organism. Objective: To analyze the bioavailability of peptides in animal models after ingestion of amaranth protein isolate and to relate it to parameters associated to cholesterol metabolism. Methods: The amaranth was crushed, the flour was defatted and its protein isolated. Amaranth peptides were analysed after in vitro digestion. Two in vivo experiments were conducted: one of acute phase and one of medium duration. In the first, the amaranth protein isolate was administered to rats and the peptides in the blood were monitored for 2 hours to check for fragments that resisted gastrointestinal digestion. The in vivo experiment 2 consisted of feeding three groups of hamsters, one with a diet recommended by AIN93 (group N) and two with hypercholesterolemic diets for 21 days, containing amaranth protein as the only dietary protein (group I), compared to casein control (group H). In this experiment were analyzed in the plasma: peptides, total cholesterol and fractions; In feces: total cholesterol and bile acids; In the liver: cholesterol, total lipids, fatty acids, Hmgcr enzymatic activity, Hmgcr expression, Srebf-2, Lxr, Abca1, Abcg8 and Ampk. Results and discussion: Peptide fragments from the in vitro digestion of amaranth protein isolate were identified and other dozens of peptide sequences were found in rats after acute amaranth administration. A higher number of peptides were found in the serum in relation to the plasma of the animals. Remarkably, ALGV peptide was found in serum of rats. This peptide is present in amaranth protein, according to databases, and is similar to fragments that present hypocholesterolemic action. In the blood of hamsters it could be found six peptides with 100 per cent coverage and similarity to the database of amaranth proteins, deserving investigation about their effects. Amaranth protein was able to suppress hypercholesterolemia when the hypercholesterolemic diet was introduced in parallel with this ingredient, with values lower for group I in 72 per cent (triglycerides), 64 per cent (total cholesterol), 80 per cent (LDL-c) in relation to the H group. A lower concentration of cholesterol and total lipids were observed in the liver of the group I compared to the H group (177 x 464 mg cholesterol / 100 g of tissue, 2.06 x 2,86 g lipids / 100 g of tissue, respectively). Lipid parameters of blood, faeces and liver were similar to those of group N, whose diet followed the recommendation for rodents. There was greater excretion of total cholesterol in group I in relation to group H, but there was no greater excretion of bile acids in feces, indicating that the effect of amaranth protein may be due to increased transintestinal cholesterol excretion, decreased micellar solubilization of cholesterol and / or modification in the expression of cholesterol transport related proteins in the intestine. There was no change in the expression of the genes analyzed in this study, but amaranth reduced the activity of the Hmgcr enzyme. It is postulated that parameters such as Ldlr expression and Acat activity are altered by amaranth intake. The fatty acid profile was also modified in order to assimilate to the N group, but inflammatory parameters related to amaranth intake should be verified due to the higher proportion of arachidonic acid in relation to the higher proportion of arachidonic acid in relation to the other groups studied. Conclusion: The bioavailability of amaranth peptides and hypocholesterolemic and hypolipidemic activity in several metabolic pathways is verified, therefore promoting cardiovascular protection

Production of marker-free transgenic rice [Oryza sativa L.] with improved nutritive quality expressing AmA1

Background: Rice seed proteins are lacking essential amino acids [EAAs]. Genetic engineering offers a fast and sustainable method to solve this problem as it allows the specific expression of heterologous EAA-rich proteins. The use of selectable marker gene is essential for generation of transgenic crops, but might also lead to potential environmental and food safety problems. Therefore, the production of marker-free transgenic crops is becoming an extremely attractive alternative and could contribute to the public acceptance of transgenic crops

Objectives: The present study was conducted to examine whether AmA1 can be expressed specifically in rice seeds, and generate marker-free transgenic rice with improved nutritive value

Materials and Methods: AmA1 was transferred into rice using Agrobacterium-mediated co-transformation system with a twin T-DNA binary vector and its integration in rice genome was confirmed by southern blot. Transcription of AmA1 was analyzed by Real-Time PCR and its expression was verified by western analysis. Protein and amino acid content were measured by the Kjeldahl method and the high-speed amino acid analyzer, respectively

Results: Five selectable marker-free homozygous transgenic lines were obtained from the progeny. The expression of recombinant AmA1 was confirmed by the observation of a 35 kDa band in SDS-PAGE and western blot. Compared to the wild-type control, the total protein contents in the seeds of five homozygous lines were increased by 1.06 tilde 12.87%. In addition, the content of several EAAs, including lysine, threonine, and valine was increased significantly in the best expressing line

Conclusions: The results indicated that the amino acid composition of rice grain could be improved by seed-specific expression of AmA1

Expression, purification and thermal stability evaluation of an engineered amaranth protein expressed in Escherichia coli

Background: The acidic subunit of amarantin (AAC)-the predominant amaranth seed storage protein-has functional potential and its third variable region (VR) has been modified with antihypertensive peptides to improve this potential. Here, we modified the C-terminal in the fourth VR of AAC by inserting four VY antihypertensive peptides. This modified protein (AACM.4) was expressed in Escherichia coli. In addition, we also recombinantly expressed other derivatives of the amarantin protein. These include: unmodified amarantin acidic subunit (AAC); amarantin acidic subunit modified at the third VR with four VY peptides (AACM.3); and amarantin acidic subunit doubly modified, in the third VR with four VY peptides and in the fourth VR with the RIPP peptide (AACM.3.4). Results: E. coli BL21-CodonPlus (DE3)-RIL was the most favorable strain for the expression of proteins. After 6 h of induction, it showed the best recombinant protein titer. The AAC and AACM.4 were obtained at higher titers (0.56 g/L) while proteins modified in the third VR showed lower titers: 0.44 g/L and 0.33 g/L for AACM.3 and AACM.3.4, respectively. As these AAC variants were mostly expressed in an insoluble form, we applied a refolding protocol. This made it possible to obtain all proteins in soluble form. Modification of the VR 4 improves the thermal stability of amarantin acidic subunit; AAC manifested melting temperature (Tm) at 34°C and AACM.4 at 37.2°C. The AACM.3 and AACM.3.4 did not show transition curves. Conclusions: Modifications to the third VR affect the thermal stability of amarantin acidic subunit.

Fuentes naturales de fitoesteroles y factores de producción que lo modifican / Natural sources of phytosterols and production factors that modify them

Actualmente los fitoesteroles son adicionados a los alimentos industrializados, lo que aumenta su costo y por lo tanto limita su consumo. Este manuscrito tiene como objetivo identificar alimentos que son una fuente natural de fitoesteroles, así como también analizar cómo el ambiente y el manejo productivo modifican su contenido, relacionando estos aspectos con la salud. Los fitoesteroles son compuestos presentes en las plantas con propiedades hipocolesterolémicas, que pueden contribuir a prevenir las enfermedades cardiovasculares. La ingesta diaria estimada de fitoesteroles varía entre 160 y 500 mg/día, sin embargo su acción beneficiosa se logra con consumos de 1500 mg a 2400 mg diarios. Las semillas de oleaginosas y de cereales son las mayores fuentes naturales de fitoesteroles. El amaranto es considerado un pseudocereal y entre los cereales tiene la mayor concentración de fitoesteroles con 178 mg/100 g, valor 5 veces superior al de la harina de trigo. El ambiente modifica el contenido de fitoesteroles de los alimentos, la sequía y temperaturas altas duplican su contenido en las semillas; la fertilización nitrogenada y la fecha de siembra pueden provocar cambios, pero se requieren más estudios al respecto. La selección de genotipos con mayor contenido de fitoesteroles y el estudio de la interacción genotipo ambiente, permitirá la identificación de aquellos genotipos con adaptación local para este carácter. El manejo productivo podrá aumentar la oferta de alimentos naturales ricos en fitoesteroles, contribuyendo a una mayor oferta de alimentos protectores de la salud(AU)

Currently phytosterols are added to processed foods, which increase their cost and therefore limits its use. This manuscript aims to identify foods that are a natural source of phytosterols, as well as analyze how the environment and production management modify its content, linking these aspects with health. Phytosterols are compounds found in plants with hypocholesterolemic properties, which may help prevent cardiovascular diseases. The estimated daily intake of phytosterols varies between 160 and 500 mg/day, but its beneficial effect is achieved with 1,500 to 2,400 mg daily consumption. Oil and cereal seeds are the largest natural sources of phytosterols. Amaranth is considered a pseudocereal and among the cereals has the highest concentration of phytosterols with 178 mg/100 g, value 5 times higher than wheat flour. Environment modifies phytosterol content in food, drought and high temperatures can double their content in seeds; nitrogen fertilization and planting date can cause changes, but more studies are needed. The selection of genotypes with higher phytosterol content and studying the genotype x environment interaction, allow the identification of those genotypes with local adaptation for this character. The production management will increase the supply of natural foods rich in phytosterols, contributing to a greater supply of health protective foods(AU)

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

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

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

Modulation of Melanin Synthesis by Amaranthus spp. L Seed Extract in Melan-a Cells

Anti-melanogenic effects of amaranth (AT), one of the key source of squalene, were investigated in melanocytes. Amaranth seed powder was extracted with water and melan-a cells were treated with various concentrations of AT. By using HPLC, content of myo-inositol, one of potential active components, was measured in the crude extract of AT.AT reduced the melanin content in melan-a melanocytes and down-regulated melanogenic enzyme activity such as tyrosinase, TRP-1 and TRP-2. By regulating melanogenic enzyme activity, AT may be a potential natural source for whitening agent. Myo-inositol was detected in AT by HPLC and may be one of the active compounds from AT involved in the regulation of anti-melanogenesis. In this study, we demonstrated that AT has anti-melanogenesis properties. This new function of amaranth may be useful in the development of new skin-whitening products and its value as food.

Sensorial and physicochemical qualities of pasta prepared with amaranth / Qualidade sensorial e físico-química de macarrão elaborado com amaranto

Current assay analyzed the sensorial acceptability of pasta when amaranth flour (AF) is added at different percentages. The physical and chemical composition and the acceptance similarities of standard formulation and formulation with a greater level of AF addition were assessed. Five formulations of pasta were used: F1 standard (0% AF); F2 (20% AF); F3 (25% AF); F4 (30% AF); F5 (35% AF). Fifty-six untrained tasters, from both genders and aged between 17 and 27 years, participated in the sensorial analysis. The physicochemical analyses determined moisture content, ash, protein, fat, carbohydrates, crude fiber, and calories. F4 with the highest AF content obtained acceptance similar to the standard formulation in all attributes. In addition, F4 provided higher levels of dietary fiber, proteins, ash, calories and lipids than standard formulation. Only carbohydrates levels in F4 were lower. Current study demonstrated that pasta with the addition of up to 30% of AF was the most sensory-accepted among the AF-added pastas. Since it provided sensory acceptance similar to the standard product, good marketing expectations are given.

Objetivou-se verificar a aceitabilidade sensorial de macarrão adicionado de farinha de amaranto (FA) em diferentes porcentagens, determinando a composição físico-química da formulação padrão e daquela com maior nível de adição de FA e aceitação semelhante. Foram elaboradas cinco formulações de macarrão sendo: F1 padrão (0% de FA) e as demais contendo 20% (F2), 25% (F3), 30% (F4) e 35% (F5) de FA. Participaram da análise sensorial 56 provadores não treinados, de ambos os gêneros, com idade entre 17 e 27 anos. Nas análises físico-químicas foi determinado o teor de umidade, das cinzas, das proteínas, dos lipídios, dos carboidratos, da fibra bruta e do valor calórico. A amostra F4 foi aquela com o maior teor de FA e com aceitação semelhante à padrão, em todos os atributos avaliados. Os teores de fibra alimentar, de proteínas, de cinzas, de calorias e dos lipídeos foram maiores em F4, obtendo-se maiores quantidades de carboidratos na formulação F1. A elaboração dos produtos permitiu comprovar que o nível de adição de até 30% de FA em macarrão foi o mais aceito pelos provadores, dentre aqueles contendo este ingrediente, obtendo-se aceitação sensorial semelhante ao produto padrão, com boas expectativas de comercialização.

Compuestos bioactivos derivados de amaranto y quinua / Bioactive components derived from amaranth and quinoa

El amaranto y la quinua son pseudocereales reconocidos por la FAO como cultivos importantes en la soberanía alimentaria para la humanidad por sus propiedades nutricionales. Poseen una alta concentración de proteínas las cuales son consideradas de alto valor nutritivo por contener aminoácidos esenciales como la lisina, el triptófano y la metionina. Se han descrito diferentes actividades biológicas de componentes aislados de amaranto y quinua como actividad antibacteriana, antitumoral, antioxidante, antiin amatoria y antihipertensiva, demostrando que tanto el amaranto como la quinua pueden ser una buena fuente de compuestos bioactivos.

Amaranth and quinoa are pseudo cereals recognized by the FAO as important crops in food sovereignty for humanity due to their nutritional properties. They have a high concentration of proteins, which are considered highly nutritious as containing essential amino acids such as lysine, tryptophan and methionine. Different biological activities isolated from amaranth and quinoa have been described as antibacterial, antitumor, antioxidant, anti-in ammatory and antihypertensive activity components, demonstrating that both amaranth and quinoa can be a good source of bioactive components.

Efeito da competição de plantas daninhas na cultura do feijoeiro / Effect of weed competition in bean

Foram realizados dois ensaios com o objetivo de avaliar o efeito da interferência de Brachiaria plantaginea (Link) Hitchc. , Amaranthus hybridus L. e Bidens pilosa L., bem como, a interferência do feijoeiro sobre o crescimento dessas plantas daninhas, além do efeito da mistura herbicida, fomesafen+fluazifop-p-butyl, sobre o desenvolvimento do feijoeiro. Os experimentos foram conduzidos em casa de vegetação, em vasos de 15 dm³, utilizandose o delineamento em blocos casualizados. No primeiro experimento avaliou-se a interferência das plantas daninhas e o efeito da mistura herbicida sobre a cultura, já no segundo avaliou o efeito da interferência do feijoeiro no desenvolvimento das três espécies de plantas daninhas. A competição do feijoeiro com as plantas daninhas ocasionou efeitos negativos no crescimento da cultura, assim como, reduziu o crescimento das plantas infestantes. B. plantaginea e B. pilosa foram às espécies daninhas que mais acarretaram efeito negativo no crescimento da cultura. Todas as plantas infestantes tiveram o seu crescimento afetado negativamente quando em convivência com a cultura. A mistura herbicida ocasionou redução no crescimento radicular da cultura, podendo se refletir em futuras perdas de rendimento em condição de déficit hídrico ou escassez de nutrientes.

The objective of this work was to evaluate the effect of interference Brachiaria plantaginea, Bidens pilosa and Amaranthus hybridus in the growth of bean plants, as well as the interference of bean on the growth of these weeds, plus the effect of the herbicide mixture, fomesafen + fluazifop-p-butyl, on the development of bean. The experiments were conducted in a greenhouse in pots of 15 dm ³, using a randomized block design. The first experiment evaluated the effect of weed and the effect of the herbicide mixture on the culture, in the second evaluated the effect of interference of bean in the development of three weed species. The bean competition with weeds caused negative effects on crop growth as well as reduced the growth of weeds. B. plantaginea and B. pilosa were the weeds that have resulted in more negative effect on crop growth. All weeds had negative affected growth of the culture. The herbicide mixture caused reduction in root growth of culture; this fact may be reflected in future income losses in water stress conditions or lack of nutrients.