Displacers improve the selectivity of phosphopeptide enrichment by metal oxide affinity chromatography / Uso de desplazadores para mejorar la selectividad de los fosfopéptidos enriquecidos por cromatografía de afinidad con óxidos de metales

Abstract: Background: A key process in cell regulation is protein phosphorylation, which is catalyzed by protein kinases and phosphatases. However, phosphoproteomics studies are difficult because of the complexity of protein phosphorylation and the number of phosphorylation sites. Methods: We describe an efficient approach analyzing phosphopeptides in single, separated protein by two-dimensional gel electrophoresis. In this method, a titanium oxide (TiO2)-packed NuTip is used as a phosphopeptide trap, together with displacers as lactic acid in the loading buffer to increase the efficiency of the interaction between TiO2 and phosphorylated peptides. Results: The results were obtained from the comparison of mass spectra of proteolytic peptides of proteins with a matrix-assisted laser desorption-ionization-time of flight (MALDI-TOF) instrument. Conclusions: This method has been applied to identifying phosphoproteins involved in the symbiosis Rhizobium etli-Phaseolus vulgaris.

Resumen: Introducción: Un proceso clave en la regulación celular es la fosforilación de proteínas, que se lleva a cabo por cinasas y fosfatasas. Sin embargo, los estudios de fosfoproteómica son difíciles debido a la complejidad de la fosforilación proteica y el número de sitios de fosforilación. Métodos: En el presente trabajo se describe una eficiente estrategia metodológica para analizar fosfopéptidos de proteínas separadas mediante electroforesis bidimensional. En este método, una columna con microesferas de dióxido de titanio (TiO2/NuTip) se utilizó para atrapar los fosfopéptidos en la superficie del TiO2 previamente empacado en una punta. El uso de desplazadores en el buffer de carga, como el ácido láctico, mejoró significativamente la selectividad. Resultados: Los resultados se obtuvieron mediante la comparación de los espectros de masas de péptidos proteolíticos de proteínas analizados utilizando un instrumento de desorción/ionización láser asistida por matriz-tiempo de vuelo (MALDI-TOF). Conclusiones: Este método se ha aplicado para la identificación de fosfoproteínas involucradas en la simbiosis del Rhizobium etli con Phaseolus vulgaris.

Efeitos da faseolamina, farinhas de feijão cru e cozido no perfil nutricional e bioquímico de ratos jovens saudáveis / Effects of phaseolamine, white bean raw and baked flours on the nutritional status and biochemical profile of healthy young rats

OBJECTIVE: To evaluate the effect of phaseolamine, white bean raw and baked flours on the nutritional status and biochemical profile of young healthy rats. Methods: We used 28 male rats, divided into 4 groups, receiving the following diets: C - standart diet; FA - standard diet added with phaseolamine (9g/kg of body weight); FF - standard diet with white bean raw flour (9g/kg of body weight); and FC - standard diet with white bean baked flour (9g/kg of body weight). The animals were kept in a controlled environment, receiving their respective diets and water ad libitumfor 28 days. The following variables were taken weekly: body weight, food and water intakes, urinary and fecal excretion. At the end of the study, blood collection was carried out for biochemical (glucose, lipid, liver and kidney profiles) and organ determinations for weight comparison between groups. RESULTS: It was possible to verify that the use of white bean raw flour, white bean baked flour, or phaseolamine showed no significant changes in the results of any of the variables analyzed. CONCLUSIONS: The use of phaseolamine and white bean raw and baked flours has no effects on the nutritional status and biochemical profile of young healthy rats

OBJETIVO: Avaliar o efeito da faseolamina, farinhas de feijão branco cru e cozido no perfil nutricional e bioquímico de ratos jovens saudáveis. MÉTODOS: Foram utilizados 28 ratos, machos, distribuídos em quatro grupos, recebendo: C ­ dieta padrão; FA ­ dieta padrão e acréscimo de faseolamina (9g/kg de peso corporal); FF ­ dieta padrão com farinha de feijão branco cru (9g /kg de peso corporal); FC ­ dieta padrão com farinha de feijão branco cozido (9g/kg de peso corporal). Os animais permaneceram em ambiente controlado, recebendo a respectiva dieta e água ad libitum por 28 dias. Foram coletados semanalmente peso corporal, consumo alimentar, ingestão hídrica, excreção urinária e fecal. Ao final do estudo realizou-se coleta de sangue para determinações bioquímicas (perfil glicídico, lipídico, hepático e renal) e de órgãos para comparação do peso entre grupos. RESULTADOS: Verificou-se que o uso da farinha de feijão branco cru, farinha de feijão branco cozido ou faseolamina não resultou em alterações significativas em quaisquer das variáveis analisadas. CONCLUSÕES: A utilização de faseolamina e farinha de feijão branco cru não possui efeitos sobre o estado nutricional e bioquímico de ratos jovens saudáveis

Bioatividade de extratos aquosos de plantas medicinais em sementes de feijão-fava / Behavioral and physiological health of lima bean seeds subjected to aqueous extract of medicinal plants

O objetivo do presente estudo foi avaliar a bioatividade de extratos aquosos de plantas medicinais em sementes de Phaseolus lunatus L. (feijão-fava) via comportamento fisiológico e fitossanitário. Foram utilizadas sementes de feijão-fava da variedade Anduzinha tratadas com seis extratos aquosos de plantas medicinais a 5% (Ocimun gratissimum, Plectranthus neachilus, Vernonia condensata, Cymbopogom citratus, Equisetum sp., e Piper aduncum L.), juntamente com a testemunha (água destilada). A bioatividade foi determinada pelo comportamento fisiológico e sanitário avaliados por meio dos testes de germinação, primeira contagem de germinação, índice de velocidade de germinação, comprimento de raiz na primeira e última contagem, e teste de sanidade. Realizou-se a análise de variância e teste Tukey a 5% de probabilidade. O extrato aquoso de cavalinha (Equisetum sp.) promoveu a melhor qualidade fisiológica das sementes de feijão-fava. Houve maior incidência de fungos nas sementes de feijão-fava que receberam o extrato de boldinho (Plectranthus neachilus).

The aim of this study was to evaluate the bioactivitt of medicinal lants aqueous extracts in Phaseolus lunatus L. (lima bean) s via through physiological and behavioral health. We the llima bean seeds of the variety Anduzinha ins of medicinal lants aqueous extracs ato 5% (Ocimun gratissimum, Plectranthus neachilus, Vernonia condensata, Cymbopogom citratus, Equisetum sp. and Piper aduncum L.) together with the control (distilled water). The bioactivity was determined by physiological and sanitary qutyies reviews through germination, first counting of germination, speed of germination, root length in the first and last counting, and sanity check. We carried out the analysis of variance and the Tukey test at 5% probability. The aqueous extract of Equisetum sp. promoted beseed physiological quality of the lima bean seed. There was a higher incidence of fungs of lima bean sract that received the extract of Plectranthus neacilus).

Repelência e atividade inseticida de pós vegetais sobre Zabrotes subfasciatus Boheman em feijão-fava armazenado / Repellency and insecticidal activity of plant powders on Zabrotes subfasciatus (Boh. ) on stored lima beans

O objetivo desta pesquisa foi avaliar o potencial inseticida de Piper nigrum L. (pimenta do reino), Ruta graveolens L. (arruda), Laurus nobilis L. (louro), Syzygium aromaticum (L.) Merr. & L.M.Perry (cravo da índia), Chenopodium ambrosioides L. (mastruz), Piper tuberculatum Jaqc. (pimenta de macaco), Tagetes erecta L. (cravo de defunto), Cymbopogon nardus L. (citronela) e Melissa officinalis L. (erva cidreira) sobre Z. subfasciatus Boh. através dos testes de confinamento, onde os insetos foram confinados durante cinco dias em um recipiente contendo dez grãos de feijão-fava com 0,3 g do pó da planta a ser testada, e, com chance de escolha por meio de uma arena circular, os insetos tiveram a possibilidade de escolha entre grãos com os pós e a testemunha, grãos sem pó das plantas. Observou-se que houve plantas que atuaram como inseticida, outras que repeliram o inseto e não causaram a morte, e outras, que além de repelir também mataram os insetos quando em contato (cravo da índia e matruz). Concluímos que: P. nigrum, P. tuberculatum, S. aromaticum e C. ambrosioides são tóxicas à Z. subfasciatus causando-lhes a morte, L. nobilis, T. erecta, e C. nardus não apresentaram efeito tóxico sobre Z. subfasciatus, C. ambrosioides, S. aromaticum, e C. nardus são repelentes à Z. subfasciatus; P. nigrum, P. tuberculatum, e T. erecta são neutras, e que M. officinalis não apresentou nenhum efeito sobre Z. subfasciatus em relação aos parâmetros avaliados.

The purpose of this research was to evaluate the insecticidal potential of Piper nigrum L., Ruta graveolens L., Laurus nobilis L., Syzygium aromaticum Merr. & L.M.Perry., Chenopodium ambrosioides L., Piper tuberculatum Jaqc., Tagetes erecta L., Cymbopogon nardus L. and Melissa officinalis L. against Zabrotes subfasciatus by free choice, in which, by means of a circular arena, insects had a choice, treatments with or without powder and no choice tests, where insects were confined for five days in a container containing ten grains of lima beans with 0.3 g of powder to be tested. It was observed that some plants have killed the insect while others just repelled the insect, causing no harm to it. Also, one of the studied plants repelled and killed the insects (S. aromaticum, C. ambrosioides and P. tuberculatum). We can conclude that: P. nigrum, P. tuberculatum, S. aromaticum e C. ambrosioides are toxic to the Z. subfasciatus, causing its death. L. nobilis, T. erecta e C. nardus do not present any toxic effect over Z. subfasciatus. C. ambrosioides, S. aromaticum e C. nardus are repellent to Z. subfasciatus. P. nigrum, P. tuberculatum e T. erecta are neutral and M. officinalis have presented no effects over Z. subfasciatus in relation to the parameters evaluated.

Respuesta glicémica al consumo de una barra de cereales-leguminosa (Phaseolus vulgaris) en individuos sanos / Glycemic response to consumption of a cereals and legume (Phaseolus vulgaris) bar on healthy individuals

El objetivo de la presente investigación fue formular una barra de cereales y leguminosa (Phaseolus vulgaris) y evaluar su impacto sobre la respuesta glicémica de individuos sanos, a fin de contribuir con la oferta de alimentos beneficiosos para la salud del consumidor. Para esto se formuló una barra con una mezcla de cereales (maíz y avena) y dos porcentajes (20% y 30%) de Phaseolus vulgaris. Adicionalmente, se preparó una barra de cereales sin leguminosa (barra control). Mediante evaluación sensorial, se seleccionó la barra con 30% de Phaseolus vulgaris, por ser evaluada con mejor sabor y textura, además esa combinación de cereales y leguminosas favorece la complementación aminoacídica y los criterios de formulación previamente establecidos. La caracterización química indicó un contenido de proteínas mayor en la barra con 30% de Phaseolus vulgaris (13,55%), en relación a la barra control (8,5%). Los contenidos de grasa, cenizas y fibra dietética no presentaron diferencias (p>0,05) entre ambas barras evaluadas. Sin embargo, el contenido de fibra soluble y de almidón resistente de la barra seleccionada resultó un 32,05% y 18,67% mayor, respectivamente, que en la barra control, con lo cual se podría contribuir con la disminución de la velocidad de absorción de la glucosa. La barra seleccionada presentó un índice glicémico bajo (49) y carga glicémica intermedia (12,0) en voluntarios sanos, lo que podría traducirse en una posible reducción de la tasa de absorción de glucosa al torrente sanguíneo, asociada a un contenido de carbohidratos de absorción lenta. La barra formulada representa una propuesta de merienda saludable para el consumidor.

The objective of this work was to formulate a cereals and legume (Phaseolus vulgaris) bar and assess its impact on the glycemic response of healthy individuals, in order to contribute to the healthy food supply beneficial to consumers. A mixture of cereals (corn and oats) and different percentages (20 and 30%) of Phaseolus vulgaris was used to formulate the bar. Additionally, a legume cereal bar without legumes (bar control) was prepared. The bar with 30% of Phaseolus vulgaris was selected through sensory evaluation, being scored with better flavor and texture. This combination of cereals and legumes aminoacid improves complementation and reaches the formulation criteria previously established. Chemical characterization indicated a higher protein content in the bar with 30% of Phaseolus vulgaris (13,55%) relative to the bar control (8,5%). The contents of fat, ash and dietary fiber did not differ between the two bars evaluated. However, the soluble fiber and resistant starch of the selected bar was a 32,05% and 18,67%, respectively, than in the control bar; this may contribute to decreasing the rate of glucose uptake. The selected bar presented a low glycemic index (49) and intermediate glycemic load (12,0) in healthy volunteers, which could lead to a possible reduction in the rate of absorption of glucose into the bloodstream, associated with a carbohydrate content of slow absorption. This bar represents a proposal of a healthy snack for the consumer.

In vitro plant regeneration system for common bean (Phaseolus vulgaris): effect of N6-benzylaminopurine and adenine sulphate / Sistema de regeneración de plantas in vitro para el frijol común (Phaseolus vulgaris): efecto del frl sulfato N6-bencilaminopurina y adenina

A method for regeneration of the commercially important common bean (Phaseolus vulgaris ) using N6-benzylaminopurine(BAP) and adenine sulphate (AS) was established. Embryogenic axes of the Costa Rican common bean cultivars Bribrí, Brunca, Guaymí, Huetar and Telire were cultured on Murashige and Skoog medium supplemented with 100 mgl-1 myo-inositol, 1 mgl-1 thiamine, 30 gl-1 sucrose, BAP (0, 5 and 10 mgl-1), AS (0, 20 and 40 mgl-1) and 8 gl-1 agar. Regardless of the concentration of BAP and AS in the induction medium, the number of shoots and leaves differed significantly among the common bean cultivars evaluated. The higher average of shoots was obtained for Brunca > Telire > Bribrí > Guaymí > Huetar. Moreover, independently of the cultivar, the induction medium supplemented with 5 mgl-1 BAP and 20 or 40 mgl-1 AS resulted in the higher average of shoots formation. Culture of Bribrí, Brunca, Guaymí, Huetar and Telire embryogenic axes on induction medium supplemented with different BAP and AS resulted in a differential response. Successful acclimatization of common bean in vitro plants were achieved in the greenhouse, and plants appeared morphologically normal. The regeneration system developed in this investigation for this important crop could be a useful tool for the genetic modification through mutagenesis or genetic transformation.

Anatomical root variations in response to water deficit: wild and domesticated common bean (Phaseolus vulgaris L)

Root anatomical responses to water deficit are diverse and regulation of water uptake strongly depends on plant anatomy. The ancestors of common bean (Phaseolus vulgaris L.) cultivars are the wild common beans. Because wild beans adapt and survive well in theon atural environment, it is hypothesized that wild common bean roots are less affected than those of domesticated beans at low substrate water potential (ψW). A wild common bean accession from Chihuahua Mexico and cv. Bayomex were studied. Seedlings with a mean root length between 3 and 4 cm were maintained for 24 h in vermiculite at ψW of -0.03 (well hydrated), -0.65, -1.48 and -2.35 MPa (partially dry). Ten anatomical characteristics of differentiation and cell division in root regions were evaluated. Thickness of epidermis and protoderm diminished similarly in wild and domesticated beans growing at low substrate ψW (between -0.65 and -2.35 MPa). At the same time, parenchymatic cell area diminished by 71 percent in the domesticated variety, but by only 32 percent in the wild bean at -2.35 MPa. Theon umber of cells in the cortex and the thickness of the xylem wall increased in both wild and domesticated beans at low substrate ψW;on evertheless, the effect was significantly lower in the wild bean. Theon umber of xylem vessels increased in the cultivar (up to 40 percent) while in the wild bean it decreased (up to 33 percent). The diameter of xylem vessels and transverse root area diminished (15 and 57 percent, respectively) in the cultivar, but in the wild common bean wereon ot affected. Anatomical root characteristics and their modifications in both differentiation and cell division in root regions demonstrated that the wild bean reacted quite differently to substrate ψW than the domesticated common bean.

Biological evaluation of the purified alpha amylase inhibitor from kidney bean [Giza 133]

The biological evaluation of the purified alpha amylase inhibitor of kidney bean [Giza 133] by using rats was carried out. Results showed that all rats gained weight at different levels. Rats in control groups gained more weight [64g] and faster than those with inhibitor. The results indicated that using 200 Ul/100g: diet was economically recommended. Diets supplemented with .200:Ul increased BV, NPU by [16%], and reduced starch hydrolysis. Glucose absorption in blood and serum insulin in all digestion hours, with little starch hydrolysis after 4 hr digestion noticed. The results also proved that using alpha amylase inhibitor in diet caused higher level of NEFA, reduction in the rate of forming fatty substances in subcutaneous layers in rats and consequently growing rate was reduced

The effects of Co2+ and Zn2+ on the contents of protein, abscisic acid, proline and chlorophyll in bean (Phaseolus vulgaris cv. Strike) seedlings.

17-day-old bean plants (Phaseolus vulgaris cv. Strike) were used to analyze the effects of Co2+ and Zn2+ on the time course of proline, total protein, chlorophyll and abscisic acid (ABA) levels in leaves. Controls, Co2+ and Zn2+-treated plants were grown for 8 days in Hoagland solution. Samples were taken at 2 day intervals. Proline, chlorophyll (a+b) and total protein contents of 17 day old primary leaves were determined by a spectrophotometer. ABA contents in roots and leaves of the seedlings were measured by high-pressure liquid chromatography. The presence of Zn2+ and Co2+ significantly increased the ABA contents in roots and leaves (p < 0.05 or p < 0.01). The increase of the abscisic acid content in the leaves was related to the content of the roots. This was further substantiated by enhanced accumulation of proline in the leaves of seedlings exposed to zinc and cobalt. The contents of chlorophyll (a+b) and total protein decreased with the concentration of both metals (p < 0.05 or p < 0.01). Cobalt proved to be comparatively more toxic than zinc.

Increase in NADH-glutamate dehydrogenase activity by mercury in excised bean leaf segments.

Application of Hg to excised bean leaf segments increased the glutamate dehydrogenase (NADH-GDH) activity substantially. However, specific activity of the enzyme decreased at lower concentration of Hg, and increased to lesser extent at higher concentration of Hg. Mercury supply increased the glutamate synthase (NADH-GOGAT) activity also. Mercury supply increased the NADH-GDH activity in the presence of NH4NO3, but to a lesser extent than in the absence of NH4NO3. The specific activity of the enzyme decreased considerably at lower concentration of Hg, but increased significantly at higher concentration of Hg. An increase in NADH-GOGAT activity was observed in the presence of NH4NO3, but specific activity of the enzyme decreased marginally. Increase in GDH activity due to Hg remained unaffected by the supply of sucrose, but was reduced by glutamine and glutathione and enhanced by Al. The glutamate dehydrogenase (+Hg enzyme) from mercury treated leaf segments had higher value of S0.5 for NADH than the enzyme (-Hg enzyme) from material not treated with mercury indicating that Hg binding to enzyme prevented NADH binding to the enzyme possibly at thiol groups. However, + Hg enzyme has more reactivity, as apparent Vmax value was higher for it. It has been suggested that Hg activates the NADH-GDH enzyme in the bean leaf segments by binding to thiol groups of protein and pronounced increase in activity by Hg suggests a possible role of enzyme under Hg-stress.

Phytostabilisation of tailing dams: a quantification of physiological and biological responses in plants.

Effect of unamended and variously amended tailings of Rajpura-Dariba mines, Udaipur, India was studied on certain physiological and biochemical responses of plants. Plants grown in unamended tailings, showed reduction in shoot-root length, shoot-root dry weight, chlorophyll content and an increase in total phenol contents and peroxidase activity. Ameliorations resulted in an increase in growth and chlorophyll contents, a decrease in total phenol contents and reduction in peroxidase activity in the test plants.