ABSTRACT
Introdução: A diabetes mellitus (DM) é uma doença crônica não transmissível importante e crescente problema de saúde pública no mundo. Mudanças no estilo de vida, como um hábito alimentar saudável, contribuem para redução da glicemia e controle do diabetes. O café é um alimento amplamente consumido e rico em compostos fenólicos com propriedades antioxidantes, com estudos sugerindo que seu maior consumo pode estar associado a um menor risco de mortalidade no diabetes. Objetivos: Analisar os efeitos das bebidas à base de café em cápsula em enzimas do metabolismo glicídico e captação de glicose em modelo de células intestinais Caco-2. Métodos: As amostras de bebidas foram preparadas com cápsulas de café espresso regular e descafeinado com ou sem adição de leite. Essas bebidas à base de café foram submetidas à digestão in vitro e seus compostos fenólicos foram quantificados e identificados. Foram realizados ensaios de permeação e quantificação de glicose nas células Caco-2, ensaios da inibição da α-glicosidase, inibição α-amilase e inibição de dipeptidil peptidase-IV (DPP-IV). Análises da capacidade antioxidante foram realizadas por meio de ensaio da capacidade de absorbância de radical oxigênio (ORAC), inibição da peroxidação lipídica (TBARS) e, também foram analisados kits comerciais o ensaio da catalase e atividade antioxidante total (TAC). Os resultados foram expressos como média e desvio padrão. Resultados Não houve diferença estatística (p>0,05) na permeação de glicose entre as diferentes bebidas de café nas células Caco-2. Na análise da capacidade de inibição da enzima α-amilase o café regular apresentou melhor inibição na fase oral, e na fase intestinal o café descafeinado apresentou melhor resultado. A inibição da e α-glicosidase os cafés descafeinado e puro foram mais efetivos. Quanto à atividade da enzima catalase na porção apical, as menores concentrações de café regular e descafeinado foram mais efetivas. A melhor capacidade antioxidante foi observada no café descafeinado. Leite e cafeína foram efetivos em estimular a enzima DPPIV. Conclusão: Todas as bebidas apresentaram capacidade antioxidante, onde se destaca a superior capacidade antioxidante do café descafeinado. As bebidas puras foram mais efetivas para inibição das enzimas α-amilase e α-glicosidase após digestão e nas células Caco-2, leite e cafeína foram melhor ativadores de DPPIV.
Background: Diabetes mellitus (DM) is an important chronic non-communicable disease and a growing public health problem worldwide. Lifestyle changes, such as healthy eating habits, contribute to lowering blood glucose levels and controlling diabetes. Coffee is a widely consumed food rich in phenolic compounds with antioxidant properties, with studies suggesting that its higher consumption may be associated with a lower risk of mortality in diabetes. Aims: To analyze the effects of capsule coffee drinks on enzymes of glucose metabolism and glucose uptake in a Caco-2 intestinal cell model. Methods: The beverage samples were prepared with espresso and decaffeinated coffee capsules with or without added milk. These coffee drinks were subjected to in vitro digestion and their phenolic compounds were quantified and identified. Glucose permeation and quantification tests were carried out on Caco-2 cells, as well as α-glucosidase inhibition, α-amylase inhibition and dipeptidyl peptidase-IV (DPP-IV) inhibition tests. Antioxidant capacity analyses were carried out using the oxygen radical absorbance capacity (ORAC) assay, lipid peroxidation inhibition (TBARS), and the catalase assay and total antioxidant activity (TAC) were also analyzed using commercial kits. The results were expressed as mean and standard deviation. Results: There was no statistical difference (p>0.05) in glucose permeation between the different coffee drinks in Caco-2 cells. In the analysis of the ability to inhibit the α-amylase enzyme, regular coffee showed better inhibition in the oral phase, and decaffeinated coffee showed better results in the intestinal phase. Decaffeinated and pure coffees were more effective at inhibiting α-glucosidase. As for the activity of the enzyme catalase in the apical portion, the lower concentrations of normal and decaffeinated coffee were more effective. The best antioxidant capacity was observed in decaffeinated coffee. Milk and caffeine were effective in stimulating the DPPIV enzyme. Conclusion: All the beverages showed antioxidant capacity, with the superior antioxidant capacity of decaffeinated coffee standing out. The pure drinks were more effective in inhibiting the enzymes α-amylase and α-glucosidase after digestion and, in Caco-2 cells, milk and caffeine were better activators of DPPIV.
Subject(s)
Coffee , Diabetes Mellitus , alpha-Amylases , Phenolic Compounds , Dipeptidyl-Peptidase IV Inhibitors , Noncommunicable Diseases , Glycoside Hydrolases , AntioxidantsABSTRACT
α-amylase is an endonucleoside hydrolase that hydrolyzes the α-1, 4-glycosidic bonds inside polysaccharides, such as starch, to generate oligosaccharides, dextrins, maltotriose, maltose and a small amount of glucose. Due to the importance of α-amylase in food industry, human health monitoring and pharmaceuticals, detection of its activity is widely required in the breeding of α-amylase producing strains, in vitro diagnosis, development of diabetes drugs, and the control of food quality. In recent years, many new α-amylase detection methods have been developed with improved speed and sensitivity. This review summarized recent processes in the development and applications of new α-amylase detection methods. The major principle of these detection methods were introduced, and their advantages and disadvantages were compared to facilitate future development and applications of α-amylase detection methods.
Subject(s)
Humans , alpha-Amylases/chemistry , Polysaccharides , Oligosaccharides , Starch , MaltoseABSTRACT
Abstract: Comparative study GC - FID /M S of essential oils of fruits, leaves and roots of the endemic plant Angelica pancicii Vandas ex Velen. revealed a significant difference in their chemical composition. The enantiomeric purity of the main component in the fruit oil (+) - ß - phellandrene was a lso confirmed. In addition, imperatorin, isoimperatorin, oxypeucedanin, oxypeucedanin hydrate, angeloylpangelin and umbelliprenin were isolated from the fruit hexane extract. The content of these coumarins in the hexane extracts from different plant parts was further determined by HPLC. The essential oils and hexane extracts were assessed for their antioxidant potential and inhibitory effect towards ï¡ - amylase and acetylcholinesterase enzymes. The fruit and leaf essential oils (> 80%) as well as the fruit he xane extract (> 62%) significantly inhibited acetylcholinesterase enzyme. Distinguish free radical scavenging properties were detected for the leaf (Inh. 95.0 ± 2.2 %) and the root (Inh. 66.0 ± 2.4 %) extracts.
Resumen: Estudio comparativo GC - FID / MS de aceites esenciales de frutas, hojas y raíces de la planta endémica Angelica pancicii Vandas ex Velen revelaron una dife rencia significativa en su composición química. También se confirmó la pureza enantiomérica del componente principal del aceite de fruta (+) - ß - felandreno. Además, se aislaron imperatorina, isoimperatorina, oxipeucedanina, hidrato de oxipeucedanina, angeloi lpangelina y umbeliprenina del extracto de hexano del fruto. El contenido de estas cumarinas en los extractos de hexano de diferentes partes de la planta se determinó adicionalmente mediante HPLC. Los aceites esenciales y extractos de hexano se evaluaron p or su potencial antioxidante efecto inhibidor de las enzimas - ï¡ - amilasa y acetilcolinesterasa. Los aceites esenciales de frutas y hojas (> 80%), así como el extracto de hexano de frutas (> 62%) inhibieron significativamente la enzima acetilcolinesterasa. Se detectaron propiedades de captación de radicales libres diferenciadas para los extractos de hoja (Inh. 95,0 ± 2,2%) y de raíz (Inh. 66,0 ± 2,4%).
Subject(s)
Acetylcholinesterase/chemistry , Angelica/chemistry , alpha-Amylases/chemistry , Oils, Volatile/chemistry , Cholinesterase Inhibitors/chemistry , Plant Leaves/chemistry , AntioxidantsABSTRACT
The antioxidant activity and the inhibitory potential of α-amylase of lyophilized hydroethanolic extracts of Conocarpus erectus leaves obtained by ultrasonication were determined. The most potent extract was subjected to ultra-high performance liquid chromatography system equipped with mass spectrometer for metabolite identification. The identified metabolites were docked in α-glucosidase to assess their binding mode. The results revealed that 60% ethanolic extract exhibited highest ferric reducing antioxidant power (4.08 ± 0.187 mg TE/g DE) and α-amylase inhibition (IC50 58.20 ± 1.25 µg/mL. The metabolites like ellagic acid, 3-O-methyl ellagic acid, ferujol, 5, 2 Ì-dihydroxy-6,7,8-trimethyl flavone and kaempferol glucoside were identified in the extract and subjected to molecular docking studies regarding α-amylase inhibition. The comparison of binding affinities revealed 3-O-methyl ellagic acid as most effective inhibitor of α-amylase with binding energy of -14.5911 kcal/mol comparable to that of acarbose (-15.7815 kcal/mol). The secondary metabolites identified in the study may be extended further for functional food development with antidiabetic properties.
Se determinó la actividad antioxidante y el potencial inhibidor de la α-amilasa de extractos hidroetanólicos liofilizados de hojas de Conocarpus erectus obtenidos por ultrasónicación. El extracto más potente se sometió a un sistema de cromatografía líquida de ultra alto rendimiento equipado con un espectrómetro de masas para la identificación de metabolitos. Los metabolitos identificados se acoplaron en α-glucosidasa para evaluar su modo de unión. Los resultados revelaron que el extracto etanólico al 60% exhibió el mayor poder antioxidante reductor férrico (4.08 ± 0.187 mg TE/g DE) e inhibición de la α-amilasa (IC50 58.20 ± 1.25 µg/mL. Los metabolitos como el ácido elágico, 3-O-metil elágico ácido, ferujol, 5, 2 Ì-dihidroxi-6,7,8-trimetil flavona y kaempferol glucósido se identificaron en el extracto y se sometieron a estudios de acoplamiento molecular con respecto a la inhibición de la α-amilasa. La comparación de las afinidades de unión reveló 3-O-metil El ácido elágico como inhibidor más eficaz de la α-amilasa con una energía de unión de -14,5911 kcal/mol comparable a la de la acarbosa (-15,7815 kcal/mol). Los metabolitos secundarios identificados en el estudio pueden ampliarse aún más para el desarrollo funcional de alimentos con propiedades antidiabéticas.
Subject(s)
Plant Extracts/chemistry , alpha-Amylases/antagonists & inhibitors , Myrtales/chemistry , Antioxidants/chemistry , Benzopyrans/analysis , In Vitro Techniques , Plant Extracts/pharmacology , Plant Leaves/chemistry , Molecular Docking Simulation , Antioxidants/pharmacologyABSTRACT
Resumo Objetivo Identificar os principais biomarcadores salivares descritos, assim como as técnicas empregadas para coleta das amostras de saliva, em estudos relacionados à avaliação da dor em pacientes submetidos a procedimentos dolorosos ou portadores de patologias dolorosas. Métodos Revisão integrativa da literatura, realizada pelas buscas bibliográficas nas bases Biblioteca Virtual em Saúde (BVS), MEDLINE/PubMed, CINAHL e EMBASE, com recorte temporal de 2009 a 2019 e período de coleta de dados entre outubro e novembro de 2019. Foram utilizados Descritores em Saúde (DeCs)e Medical SubjectHeadings (MeSH), para responder à pergunta norteadora: Quais são e como são utilizados os biomarcadores salivares na avaliação da dor? Foi realizada uma análise descritiva dos artigos, sendo os dados extraídos e registrados em uma planilha desenvolvida para o presente estudo. Resultados Das 126 publicações identificadas, 22 artigos foram incluídos para a análise. Constatou-se que os artigos são, majoritariamente, desenvolvidos com adultos durante realização de procedimentos dolorosos ou portadores de patologias dolorosa. Os principais biomarcadores salivares avaliados foram a alfa-amilase e o cortisol, e as principais técnicas para coleta de saliva foram o Salivette® e a coleta passiva. Conclusão Os estudos indicam que a mensuração objetiva da dor é um desafio. Os principais biomarcadores salivares descritos são o cortisol e a alfa-amilase, sendo o Salivette®a principal técnica utilizada para coleta das amostras de saliva. A dosagem das moléculas salivares é incipiente e empregada de forma complementar na avaliação da dor em pacientes de diferentes faixas estárias, submetidos à procedimentos dolorosos ou portadores patologias dolorosas.
Resumen Objetivo Identificar los principales biomarcadores salivales descriptos, así como las técnicas utilizadas para la recolección de las muestras de saliva en estudios relacionados con la evaluación del dolor en pacientes sometidos a procedimientos dolorosos o con patologías dolorosas. Métodos Revisión integrativa de la literatura, realizada por medio de búsquedas bibliográficas en las bases Biblioteca Virtual em Saúde (BVS), MEDLINE/PubMed, CINAHL y EMBASE, con un recorte temporal del 2009 al 2019 con un período de recolección de datos de octubre a noviembre de 2019. Se utilizaron Descriptores en Salud (DeCs) y Medical SubjectHeadings (MeSH), para responder a la pregunta orientadora: ¿Cuáles son los biomarcadores salivales en la evaluación del dolor y cómo se utilizan? Se realizó un análisis descriptivo de los artículos y los datos extraídos y registrados en una planilla desarrollada para el presente estudio. Resultados De las 126 publicaciones identificadas, se incluyeron 22 artículos para análisis. Se constató que los artículos están, mayoritariamente, desarrollados con adultos durante la realización de procedimientos dolorosos o con patologías dolorosas. Los principales biomarcadores salivales evaluados fueron alfa-amilasa y cortisol, y las principales técnicas para la recolección de saliva fueron Salivette® y la recolección pasiva. Conclusión Los estudios indican que la medición objetiva del dolor es un desafío. Los principales biomarcadores salivales que se describen son el cortisol y la alfa-amilasa y Salivette® la principal técnica utilizada para la recolección de muestras de saliva. La dosificación de las moléculas salivales es incipiente y utilizada de forma complementaria en la evaluación del dolor en pacientes de distintos grupos de edad, sometidos a procedimientos dolorosos o con patologías dolorosas.
Abstract Objective To identify the main salivary biomarkers described and the techniques used for saliva sample collection in studies related to pain assessment in patients undergoing painful procedures or experiencing painful diseases Methods An integrative literature review was conducted via bibliographic searches in the Virtual Health Library (VHL), MEDLINE/PubMed, CINAHL, and EMBASE databases for the period from 2009 to 2019; data were collected in October and November 2019. The DeCs health descriptors and the Medical Subject Headings (MeSH) were used to answer the guiding question: "Which salivary biomarkers are used in pain assessment and how are they employed?" A descriptive analysis of the articles was performed; data were collected and recorded in a spreadsheet developed for the present study. Results Of the 126 published articles identified, 22 articles were included for analysis. The articles were mainly regarding adults undergoing painful procedures or patients experiencing painful diseases. The main salivary biomarkers evaluated were alpha-amylase and cortisol, and the main saliva collection techniques were Salivette® and passive collection. Conclusion The studies indicated that objective pain measurement is a challenge. The main salivary biomarkers evaluated were cortisol and alpha-amylase, and the main technique employed for saliva sample collection was Salivette®. The dosage of salivary molecules is emerging for use as a complement in pain assessment in patients of different ages undergoing painful procedures or experiencing painful diseases.
Subject(s)
Humans , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Saliva , Pain Measurement , Hydrocortisone , Biomarkers , Diagnostic Techniques and Procedures , alpha-Amylases , Evidence-Based Practice , Anti-Inflammatory AgentsABSTRACT
Aims@#The study aimed to investigate the effect of glucose on alpha-amylase and glucoamylase production in some Indonesian indigenous fungi.@*Methodology and results@#Fungi were screened for their ability to produce alpha-amylase and glucoamylase in the presence of glucose. The strains were grown in a medium containing starch and glucose as carbon sources with glucose concentrations varying from 0 to 5% for four days, and the alpha-amylase and glucoamylase were analyzed at the end of the growth period. Most strains showed repression on the amylases production when glucose was added to the medium. However, some strains showed no repression on amylases production when glucose was supplemented to the medium. The addition of glucose repressed glucoamylase production, but no repression on alpha-amylase was noted for strain KKB4, vice versa, there was repression on alpha-amylase production but no repression on glucoamylase production for strain FIG1. Strains FNCC 6151 and MLT1J1 showed no repression on both alpha-amylase and glucoamylase production when glucose was added to the medium up to 5%. The occurrence of repression in the production of alpha-amylase and glucoamylase was strain-specific.@*Conclusion, significance and impact of study@#Out of the nine indigenous fungi strains examined, strains FNCC 6151 and MLT1J1 showed no repression on both alpha-amylase and glucoamylase production when glucose was added to the medium up to 5%. Those two strains have the potential to be improved further to produce both alpha-amylase and glucoamylase.
Subject(s)
Glucosidases , alpha-Amylases , Glucan 1,4-alpha-GlucosidaseABSTRACT
Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM-465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% α-helices, 15% β-sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (∆G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.
A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (∆G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.
Subject(s)
Escherichia coli/genetics , Geobacillus , Genetic Vectors , alpha-Amylases/geneticsABSTRACT
Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% α-helices, 15% ß-sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (∆G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.
A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (∆G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.
Subject(s)
Escherichia coli/genetics , alpha-Amylases/genetics , alpha-Amylases/metabolism , Temperature , Enzyme Stability , Cloning, Molecular , Geobacillus , Hydrogen-Ion ConcentrationABSTRACT
The objective of this study was to evaluate biological and phytochemical properties of the aqueous extract from the leaves of Miconia chamissois Naudin (AEMC). Phytochemical properties were assessed by analyzing the chromatographic profile and the polyphenol content of AEMC. Biological properties evaluation was conducted based on cytotoxicity assay and by evaluating the antioxidant, antimicrobial, and enzymatic inhibition activities. Results indicated the presence of phytochemicals in AEMC such as flavonoids and polyphenols, including rutin, isoquercitrin and vitexin derivatives. AEMC showed antioxidant activity, which may be attributed to the high polyphenolic content. Moreover, AEMC demonstrated in vitro enzyme inhibition activity against tyrosinase and alpha-amylase, as well as showed low cytotoxicity. On the other hand, AEMC exhibited weak antimicrobial activity against S. aureusand C. albicans. Thus, AEMC is a promising alternative in search of potential drugs for the treatment of diseases induced by oxidative stress and inflammation, conditions due to hyperpigmentation processes, such as melisma, as well as for diabetes.
El objetivo de este estudio fue detectar las propiedades biológicas y fitoquímicos del extracto acuoso de las hojas de Miconia chamissois Naudin (AEMC). Las propiedades fitoquímicas se evaluaron analizando el perfil cromatográfico y el contenido de polifenoles de AEMC. La evaluación de las propiedades biológicas se realizó en base al ensayo de citotoxicidad y evaluando las actividades de inhibición antioxidante, antimicrobiana y enzimática. Los resultados indicaron la presencia de fitoquímicos en AEMC, como flavonoides y polifenoles, que incluyen derivados de rutina, isoquercitrina y vitexina. AEMC mostró una actividad antioxidante considerable, que puede atribuirse al alto contenido polifenólico. Además, AEMC exhibió actividad de inhibición enzimática in vitro contra tirosinasa y alfa-amilasa, así como mostró baja citotoxicidad. Por otro lado, AEMC demostró actividad antimicrobiana débil contra S. aureusy C. albicans. Por lo tanto, AEMC es una alternativa prometedora en busca de posibles drogas para el tratamiento de enfermedades inducidas por el estrés oxidativo y la inflamación, afecciones debidas a procesos de hiperpigmentación, como el melasma, así como para la diabetes.
Subject(s)
Plant Extracts/pharmacology , Plant Extracts/chemistry , Melastomataceae/chemistry , Flavonoids/analysis , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Monophenol Monooxygenase/antagonists & inhibitors , alpha-Amylases/antagonists & inhibitors , Polyphenols/analysis , Anti-Infective Agents/pharmacology , Antioxidants/pharmacologyABSTRACT
Poor storage conditions provide favorable environment to stored grain pests for their growth. The bio-pesticides are the best alternatives to synthetic pesticides. Present study was conducted to compare toxicity of Rubus fruticosus and Valeriana jatamansi against granary weevil, Sitophilus granarius and subsequent changes in enzyme activity responsible for grain damage. In current research 5 g of R. fruticosus fruit and V. jatamansi rhizome powders were tested separately against S. granarius, in 50 g wheat whole grains for seven days in comparison with the control. The enzymatic activity of malate dehydrogenase and α-amylase was observed in the cellular extracts of S. granarius. The insects were crushed and homogenized in phosphate-buffer solution and centrifuged at 10000 rpm for 5 minutes. For the enzymatic measurement supernatant was tested; the spectrophotometer was adjusted at 340 nm. The reagents were mixed and incubated at 25 °C for five minutes. The cuvettes were placed in the experimental and reference sites of spectrophotometer and recorded the change in absorbance for 3-4 minutes. There was 5.60% and 14.92% reduction in the activity of malate dehydrogenase in R. fruticosus and V. jatamansi, treated insects, respectively. The alpha amylase enzyme activity was 6.82% reduced and 63.63% increase in R. fruticosus and V. jatamansi, treated insects, respectively. Present study addresses that both plant powders are effective against granary weevil by altering enzyme activities so both the plant powders can be used as bio-pesticides against the stored grains pests.
As más condições de armazenamento proporcionam um ambiente favorável às pragas armazenadas para o crescimento. Os biopesticidas são as melhores alternativas aos pesticidas sintéticos. O presente estudo foi conduzido para comparar a toxicidade de Rubus fruticosus e Valeriana jatamansi contra gorgulhos, Sitophilus granarius e subsequentes alterações na atividade enzimática responsáveis ââpor danos aos grãos. Na pesquisa atual, 5 g de frutos de R. fruticosus e pós de rizoma de V. jatamansi foram testados separadamente contra S. granarius, em 50 g de grãos integrais de trigo por sete dias, em comparação com o controle. A atividade enzimática da malato desidrogenase e α-amilase foi observada nos extratos celulares de S. granarius. Os insetos foram esmagados e homogeneizados em solução tampão fosfato e centrifugados a 10000 rpm por 5 minutos. Para a medição enzimática, o sobrenadante foi testado; o espectrofotômetro foi ajustado a 340 nm. Os reagentes foram misturados e incubados a 25 °C por cinco minutos. As cubetas foram colocadas nos locais experimentais e de referência do espectrofotômetro e registradas as alterações na absorbância por 3-4 minutos. Houve redução de 5,60% e 14,92% na atividade da malato desidrogenase em R. fruticosus e V. jatamansi, insetos tratados, respectivamente. A atividade da enzima alfa amilase foi reduzida em 6,82% e aumento de 63,63% em R. fruticosus e V. jatamansi, insetos tratados, respectivamente. O presente estudo aborda que ambos os pós de plantas são eficazes contra o gorgulho do celeiro, alterando as atividades enzimáticas, de modo que ambos os pós de plantas possam ser usados ââcomo biopesticidas contra pragas de grãos armazenados.
Subject(s)
Animals , Valerian/toxicity , Weevils , Biological Control Agents/administration & dosage , Rubus/toxicity , Pest Control, Biological/methods , alpha-Amylases , Food Storage/standards , Malate DehydrogenaseABSTRACT
Monosodium glutamate (MSG) is a flavor enhancer widely used in the food industry, with obesogenic properties, in addition to causing alterations in the oral cavity. The aim of the study was to observe the morphofunctional changes in the parotid gland after the administration of MSG in rats. 18 newborn male Sprague Dawley rats were used, divided into three groups (Control group; MSG1 group: 4 mg/g weight of monosodium glutamate, 5 doses, kept for 8 weeks, and MSG2 group: 4 mg/g weight of MSG, 5 doses, kept for 16 weeks). The body mass index (BMI) was calculated, and the salivary flow, pH, a-amylase activity, Na, Cl, K and Ca were analyzed by quantitative analysis. After euthanasia by ketamine/xylazine overdose, parotid volume was analyzed and stereology was performed. MSG administration caused an increase in BMI and a decrease in parotid volume as well as a reduction in salivary flow and pH and an increase in a-amylase activity, also increasing the salivary sodium and chlorine levels. Alterations in the normal stereological parameters of the gland were observed. Exposure to MSG caused morphofunctional alterations at parotid gland.
El glutamato monosódico (MSG), es un potenciador del sabor ampliamente utilizado en la industria alimentaria. Diversos estudios han propuesto la relación entre éste y el desarrollo de obesidad, además de provocar alteraciones en la cavidad oral. El objetivo del estudio fue observar los cambios morfofuncionales a nivel de la glándula parótida, posterior a la administración de MSG en ratas. Se utilizaron 18 ratas neonatas Sprague Dawley machos, divididas en tres grupos según su tiempo de exposición y dosis a MSG (Grupo Control, Grupo MSG1: 4 mg/g peso de glutamato monosódico, 5 dosis, mantenidas 8 semanas, Grupo MSG2: 4 mg/g peso de MSG, 5 dosis, mantenidas 16 semanas. Fue calculado el índice de masa corporal (BMI), además de ser analizado el flujo salival, pH, actividad de α-amilasa, y Na, Cl, K y Ca mediante análisis semicuantitativo. Luego de la eutanasia por sobredosis de ketamina/xilasina, las glándulas parótidas fueron extraídas y analizado su volumen y fueron procesadas para histología, y estudio estereológico. La administración de MSG causó aumento en BMI y disminución del volumen parotídeo, además de disminución del flujo y pH salival, así como aumento en actividad de la a-amilasa, aumentando además los niveles de sodio y cloro salival. Fueron observadas alteraciones a nivel de los parámetros estereológicos normales de la glándula. La exposición a MSG causó alteraciones morfofuncionales a nivel parotídeo, observándose una disminución del volumen de la glándula, acompañado de alteraciones en el adenómero y conductos estriados de la glándula, implicados en la producción, secreción y modificación de la saliva, la cual se vio alterada, en el flujo, pH, y en sus componentes.
Subject(s)
Animals , Male , Rats , Parotid Gland/drug effects , Sodium Glutamate/administration & dosage , Flavoring Agents/administration & dosage , Saliva/chemistry , Sodium/analysis , Sodium Glutamate/pharmacology , Time Factors , Body Mass Index , Chlorine/analysis , Analysis of Variance , Rats, Wistar , alpha-Amylases/analysis , Flavoring Agents/pharmacology , Hydrogen-Ion ConcentrationABSTRACT
Population growth has raised food production, and new sources are needed to increase quantity and quality of agricultural products. Carbamates and organophosphates are insecticide classes used worldwide as acetylcholinesterase (AChE) inhibitors. Plants have a natural resistance to insects, which can be employed in pest control as a new alternative to reduce the use of chemicals. An alternative may be the use of α-amylase inhibitors, which are digestive enzymes that impair pest species growth and development. Another would be acetylcholinesterase inhibitors since they damage the normal functioning of the central and peripheral nervous system, by releasing high concentrations of acetylcholine in cholinergic synapses. This substance accumulation increases stimulations that lead to behavioral changes, asphyxia, hyperactivity, and death. Botanical agrochemicals are believed to have advantages over synthetic ones, as they are rapidly degraded in the environment. In this scenario, plants have played an important role in pest control as sources of interest for the synthesis of new molecules for agricultural use. The present study evaluated acetylcholinesterase and α-amylase inhibition by microplate method, from leaf extracts of Mouriri elliptica Martius with different polarities.
O crescimento populacional tem aumentado a quantidade de produção alimentícia, sendo necessárias novas fontes para o aumento da quantidade e qualidade dos produtos agrícolas. Os carbamatos e organofosforados são classes de inseticidas utilizadas em todo o mundo, são inibidores da acetilcolinesterase (AChE). Os vegetais possuem uma resistência natural aos insetos, e esse método de resistência pode ser utilizado no controle das pragas como uma nova alternativa para redução do uso de inseticidas químicos, tais como os inibidores da α-amilase, enzima digestiva, a qual sua inibição prejudica o crescimento e desenvolvimento de espécies de pragas. E os inibidores da acetilcolinesterase que danificam o funcionamento normal do Sistema Nervoso Central e Periférico, através de elevadas concentrações da acetilcolina que ficam depositadas nas sinapses colinérgicas. Este acúmulo de ACh provoca uma grande estimulação que leva á alterações comportamentais, asfixia, hiperatividade e a morte. Estudos já realizados mostraram que os agrotóxicos botânicos têm vantagens sobre os sintéticos, sendo degradados rapidamente no meio ambiente. Neste cenário os vegetais têm desempenhado um importante papel no controle de pragas, através da síntese de novas moléculas para uso na agricultura. O presente trabalho avaliou a inibição das enzimas acetilcolinesterase e α-amilase através do método da microplaca a partir dos extratos das folhas de Mouriri elliptica Martius em diferentes polaridades.
Subject(s)
Acetylcholinesterase , Pest Control , Cholinesterase Inhibitors , alpha-Amylases , Pesticides , Food Production , Chromatography , Agrochemicals , Crops, Agricultural , GrasslandABSTRACT
Seed vigor is a key factor affecting seed quality. The mechanical drying process exerts a significant influence on rice seed vigor. The initial moisture content (IMC) and drying temperature are considered the main factors affecting rice seed vigor through mechanical drying. This study aimed to determine the optimum drying temperature for rice seeds according to the IMC, and elucidate the mechanisms mediating the effects of drying temperature and IMC on seed vigor. Rice seeds with three different IMCs (20%, 25%, and 30%) were dried to the target moisture content (14%) at four different drying temperatures. The results showed that the drying temperature and IMC had significant effects on the drying performance and vigor of the rice seeds. The upper limits of drying temperature for rice seeds with 20%, 25%, and 30% IMCs were 45, 42, and 38 °C, respectively. The drying rate and seed temperature increased significantly with increasing drying temperature. The drying temperature, drying rate, and seed temperature showed extremely significant negative correlations with germination energy (GE), germination rate, germination index (GI), and vigor index (VI). A high IMC and drying temperature probably induced a massive accumulation of hydrogen peroxide (H2O2) and superoxide anions in the seeds, enhanced superoxide dismutase (SOD) and catalase (CAT) activity, and increased the abscisic acid (ABA) content. In the early stage of seed germination, the IMC and drying temperature regulated seed germination through the metabolism of H2O2, gibberellin acid (GA), ABA, and α-amylase. These results indicate that the metabolism of reactive oxygen species (ROS), antioxidant enzymes, GA, ABA, and α-amylase might be involved in the mediation of the effects of drying temperature on seed vigor. The results of this study provide a theoretical basis and technical guidance for the mechanical drying of rice seeds.
Subject(s)
Abscisic Acid/metabolism , Antioxidants/pharmacology , Catalase/metabolism , Gene Expression Regulation, Plant/drug effects , Germination , Gibberellins/metabolism , Hydrogen Peroxide/chemistry , Malondialdehyde/chemistry , Oryza/metabolism , Oxygen/chemistry , Plant Proteins/genetics , Reactive Oxygen Species , Seeds/metabolism , Superoxide Dismutase/metabolism , Superoxides/chemistry , Temperature , Weather , alpha-Amylases/metabolismABSTRACT
Este estudo objetivou avaliar as possíveis associações com parâmetros objetivos, como cortisol e alfa-amilase salivares e a Variabilidade da Frequência Cardíaca (VFC) na eminência de procedimentos odontológicos. A amostra foi constituída de 30 pacientes com indicação para exodontia de terceiros molares inferiores erupcionados, caracterizada quanto ao gênero, idade, escolaridade, renda familiar, evento traumático prévio, intensidade de dor e ansiedade ao tratamento odontológico. Para avaliação do cortisol e da alfa-amilase foram utilizados kits de coleta salivar e dados da VFC, obtidos após processamento dos intervalos RR, registrados com o frequencímetro portátil Polar RS800, em três momentos: entrevista, cirurgia e pós-operatório. Foram analisadas as variáveis do domínio tempo (SDNN e rMSSD) e frequência (razão LF/HF). Nesta amostra de voluntários saudáveis, a ansiedade ao tratamento odontológico e o antecedente de experiências traumáticas não se mostraram correlacionados à maior intensidade de dor pós-operatória. Apenas três pacientes foram classificados como ansiosos e quatro relataram algum evento prévio. Estes pacientes foram reunidos em um grupo. Não houve diferença significativa entre este grupo e os não ansiosos em relação às categorias sócio demográficas e dor pós-operatória. Não houve correlação estatisticamente significativa entre as concentrações de alfa-amilase e cortisol com os valores das variáveis de VFC, nos três dias estudados. As variáveis estudadas também não expressaram alteração evolutiva com significância estatística. O estudo de marcadores não invasivos de estresse vem ganhando importância na detecção precoce das respostas autonômicas frente a situações potencialmente estressoras, como cirurgias odontológicas.
Subject(s)
Hydrocortisone , Dental Anxiety , alpha-AmylasesABSTRACT
The international standards for top glucomannan flour require a minimum of 70% glucomannan. The glucomannan content of Amorphophallus oncophyllus flour was approximately 60%, with starch as the major impurity. Elimination of starch was expected to increase the purity of glucomannan. The purpose of this research was to study starch hydrolysis of the flour using α-amylase. Temperature (35.5-84.5oC), time (0.4-3.6 h) and pH (2.2-8.8) of hydrolysis were selected as independent variables. A central composite design of response surface methodology (RSM) was performed to obtain the optimum condition. This approach was a novelty of this enzymatic purification of A. oncophyllus. Glucomannan content, starch content, and solubility were chosen as the response variables. The models were reliable for predicting the responses (R2≥ 0.771). It was predicted that the highest glucomannan content (93.0%) obtained at the lowest starch content (1.14%), which hydrolysed at pH 6.17, 84.5oC and 3.6 h. Prior the verification of the optimum hydrolysed condition from the model, the glucomannan and starch content was 81.59% and 2.27%, respectively. After purification, the absorbance of the ß-1,4 glycosidic bond increased as a sign of higher glucomannan purity. A less rough surface and irregular shape of the grain morphology was observed after purification.
Os padrões internacionais para a farinha de alta calidade de glucomanan requerem um mínimo de 70% de glucomanan. O conteúdo de glucomanano da farinha de Amorphophallus oncophyllus foi de aproximadamente 60%, com o amido como a maior impureza. Esperava-se que a eliminação do amido aumentasse a pureza do glucomanan. O objetivo desta pesquisa foi estudar a hidrólise do amido da farinha usando α-amilase. A temperature (35,5-84,5oC), o tempo (0,4-3,6 h) e o pH (2,2-8,8) da hidrólise foram selecionados como variáveis independentes. Um desenho central composto pertencente á metodologia da superfície de resposta (MSR) foi realizado para obter a condição ótima. Esta abordagem foi uma novidade desta purificação enzimática de A. oncophyllus. O conteúdo de glucomanan, conteúdo de amido e solubilidade foram escolhidos como as respostas. Os modelos foram confiáveis para predizer as respostas (R2≥ 0,771). Os modelos indicaram que o maior conteúdo de glucomanan (93,0%) foram obtidos no menor conteúdo de amido (1,14%),que hidrolisou a um pH 6,17, 84,5ºC e 3,6 h. Antes da verificação da condição hidrolisada ótima do modelo, o conteúdo de glucomanan e amido foi de 81,59% e 2,27%, respectivamente. Após a purificação, a absorbância da ligação ß-1,4 glicosídica aumentou com um sinal de maior pureza de glucomanan. Uma superfície mais lisa e forma irregular da morfologia do grão foi observada após a purificação.
Subject(s)
Amorphophallus , alpha-Amylases , Flour , HydrolysisABSTRACT
Introducción: la α-amilasa es una enzima que tiene la capacidad de digerir uniones α (1,4). En la leche sus funciones son poco conocidas. Objetivos: determinar la actividad de la α-amilasa en el suero lácteo humano. Materiales y métodos: la medición se realizó con un kit comercial para determinación de amilasa sérica y se utilizaron muestras de leche humana. Resultados: se encontró una actividad promedio de 2.000 U/l. La comparación de las medias de los valores de actividad desde el segundo al quinto día (1,372±0,796) con los correspondientes al lapso desde el sexto día al cuadragésimo séptimo (0,412±0,250) demostró que los valores eran significativamente diferentes (p<0,0025). Los coeficientes de variación de los dos grupos mencionados (1-5 días y 6-47 días) fueron 58 y 61% respectivamente. De manera adicional, la expresión de la amilasa mostró una gran variación entre madres. Conclusiones: a partir de esta información y de los antecedentes bibliográficos, la α-amilasa láctea tendría una función de defensa en el organismo del recién nacido lactante, y recién al iniciar la alimentación suplementada cumpliría el rol conocido. Aún así el significado biológico de su función en la leche es un problema abierto.
Subject(s)
Colostrum , Enzymes , alpha-Amylases , Milk, HumanABSTRACT
Salivary alpha-amylase (sAA) is a stress biomarker in human diseases, but there are no reports of sAA measurements in diseased dogs. This study measured the sAA and serum alpha-amylase (AA) levels in 16 healthy dogs and 31 diseased dogs using a kinetic enzyme assay to assess the stress status. The sAA and serum AA levels were significantly higher in the diseased dogs than in healthy dogs (p < 0.05), but there was no correlation between the 2 groups (r = 0.251, p = 0.089). This suggests that sAA can be useful as a stress biomarker in diseased dogs.
Subject(s)
Animals , Dogs , Humans , alpha-Amylases , Enzyme Assays , SalivaABSTRACT
Los inhibidores selectivos de la α-amilasa salival y pancreática humana, son un medio eficaz para controlar los niveles de azúcar en la saliva y la sangre, durante el control de la caries y la diabetes. En la presente revisión, después de identificar las principales funciones de la enzima, se discutieron exponen algunas de las respuestas observadas después de la exposición de la α-amilasa a las plantas, en escala molecular y entera. Diferentes tipos de moléculas de plantas medicinales actúan como inhibidores de la α-amilasa, como una terapia alternativa o complementaria en el tratamiento de la diabetes en y en el control de uno de los factores de la formación la caries (dieta) (AU)
Selective inhibitors of human salivary and pancreatic α-amylase are an effective means of controlling saliva and blood sugar levels in the management of caries and diabetes. In the present review, after identifying the main functions of the enzyme, it was exposed some of the observed responses after exposure to α-amylase at the molecular and whole plants scale. Different types of medicinal plants molecules have been found to act as α-amylase inhibitors, as an alternative or complementary therapy in the management of diabetes and control to one of the predisposing factors to caries (diet) (AU)
Subject(s)
Humans , alpha-Amylases , Dental Caries , Diabetes Mellitus , Enzyme Inhibitors , Plants, Medicinal , Complementary Therapies , Glycoside Hydrolase InhibitorsABSTRACT
Background: α-Amylase is widely used in the starch processing, food and paper industries, hydrolyzing starch, glycogen and other polysaccharides into glucose, maltose and oligosaccharides. An α-amylase gene family from Aspergillus niger CBS513.88 encode eight putative α-amylases. The differences and similarities, biochemical properties and functional diversity among these eight α-amylases remain unknown. Results: The eight genes were cloned and expressed in Pichia pastoris GS115 by shaking-flask fermentation under the induction of methanol. The sequence alignment, biochemical characterizations and product analysis of starch hydrolysis by these α-amylases were investigated. It is found that the eight α-amylases belonged to three different groups with the typical structure of fungal α-amylase. They exhibited maximal activities at 3040°C except AmyG and were all stable at acidic pH. Ca2+ and EDTA had no effects on the activities of α-amylases except AmyF and AmyH, indicating that the six amylases were Ca2+ independent. Two novel α-amylases of AmyE and AmyF were found. AmyE hydrolyzed starch into maltose, maltotriose and a small amount of glucose, while AmyF hydrolyzed starch into mainly glucose. The excellent physical and chemical properties including high acidic stability, Ca2+-independent and high maltotriose-forming capacity make AmyE suitable in food and sugar syrup industries. Conclusions: This study illustrates that a gene family can encode multiple enzymes members having remarkable differences in biochemical properties. It provides not only new insights into evolution and functional divergence among different members of an α-amylase family, but the development of new enzymes for industrial application.
Subject(s)
Aspergillus niger/enzymology , alpha-Amylases/genetics , alpha-Amylases/chemistry , Pichia/metabolism , Starch , Temperature , Food Industry , Cloning, Molecular , Fermentation , Hydrogen-Ion Concentration , HydrolysisABSTRACT
ABSTRACT Our present investigation deals with the phytochemical screening, estimation of total flavonoids, terpenoids and tannin contents to evaluate the anti-diabetic activities of Salacia oblonga stem followed by GC-MS analysis. It explores the natural compounds and the potential α-amylase and α-glucosidase inhibitory actions of stem extracts. The aqueous stem extract was selected from other extracts (ethanol, acetone, petroleum ether and chloroform) for the in vitro study of anti-diabetic activity by alpha amylase and alpha glucosidase inhibitory assays. The stem extract was also analyzed by gas chromatography mass spectrometry to identify the natural chemical components. Phytochemical analysis of aqueous stem extract showed major classes of secondary metabolites such as phenols, flavonoids, alkaloids, terpenoids, tannins, saponins. The total flavonoid, terpenoid, and tannin contents were quantified as 19.82±0.06 mg QE/g, 96.2±0.20 mg/g and 11.25±0.03 mg TAE/g respectively. The percentage inhibition of assays showed maximum inhibitory effects (59.46±0.04% and 68.51±0.01%) at a concentration of 100 mg/mL. The IC50 values of stem extract was found to be 73.56 mg/mL and 80.90 mg/mL for alpha amylase and alpha glucosidase inhibition. Fifteen chemical constituents were found by GC-MS analysis. This study suggest the aqueous stem extract of Salacia oblonga might be considered as potential source of bio active constituents with excellent antidiabetic activity.