RESUMO
This study aimed to determine the nutritional components (macronutrients ans minerals) and α-amylase inhibition capacity of freeze-dried grumixama (Eugenia brasiliensis Lam) seeds (S) and pulp/peel (P) portions, at ripe and mid-ripe stages. In vitro digestion was also performed on S and P from grumixama to assess the bioaccessibility of total phenolic compound (TPC), flavonoids (TFC), and anthocyanins (TAC), as well as to examine their impact on antioxidant activity (DPPH, ABTS, FRAP). The ripening process impacts the bioactive compounds and individual phenolics of S and P portions. The ripe S was source of myricetin and exhibited higher antioxidant activity, while mid-ripe S was high in flavonoids and cinnamic acid with higher antiglycemic potential. Ripe P showed higher soluble fiber, carbohydrate, TAC, and caffeic acid content, whereas mid-ripe P had increased mineral content (calcium, potassium, manganese), catechin, and TPC. After in vitro digestion, the P portion showed a bioaccessibility of total phenolic content (TPC) and total flavonoid content (TFC) exceeding 40% at intestinal phase. In contrast, the S portions had better release of TPC and TFC and antioxidant activity at gastric phase. Considering the outstanding nutritional and biological properties of grumixama fruit, freeze-dried S and P portions from both ripening stages possess could be explored as valuable sources of nutrients and antioxidant compounds.
Assuntos
Antioxidantes , Eugenia , Antioxidantes/análise , Antocianinas/análise , Frutas/química , Flavonoides/análise , Fenóis/análise , Minerais/análiseRESUMO
Red cabbage is rich in phytochemical compounds, and its consumption, either raw or cooked, has been linked to the prevention of several diseases. This work aimed to investigate the influence of cooking methods on in vitro bioaccessibility of phenolics and antioxidant activity of red cabbage. The vegetable was subjected to boiling, steaming, and microwaving for different times to evaluate color parameters, total phenolic (TPC), total flavonoid (TFC), anthocyanin content (AC), and antioxidant activity (FRAP, DPPH, and ABTS). The phytochemical bioaccessibility before and after cooking was also evaluated by in vitro simulated digestion. Steaming showed the most significant retention of the compounds after 20 and 25 min of cooking (72-86% for TPC, 72-77% for TFC, 75-79% for FRAP, 84-91% for DPPH, 70-83% for ABTS), followed by microwaving, which was more stable in 10 min. Microwaving decreased TFC and AC over time. Boiling did not show significant differences between the cooking times and showed more than 50% of losses of TPC, TFC, and AC and 30 to 60% of antioxidant activity. Steaming was the best cooking method, showing the most significant tendency to black coloration (< L*). In 10 min, it still showed the highest percentages of increase in TPC and the minor losses of TFC and AC in the gastric and intestinal phases. Steaming also increased the antioxidant after digestion when compared to uncooked red cabbage. These results are important to help consumers choose the most effective cooking method for red cabbage to retain its health-promoting components.
Assuntos
Antioxidantes , Brassica , Antioxidantes/análise , Flavonoides/análise , Brassica/química , Fenóis/análise , Antocianinas/análise , Culinária/métodos , Compostos FitoquímicosRESUMO
Fruits from Ziziphus joazeiro from Mata Paraibana (MP), Borborema (BB), Agreste (AG) and Sertão Paraibano (SP) were assessed to determine their morphological characteristics, nutritional composition, content and bioaccessibility of phenolic compounds and antioxidant activity. In general, juá fruits presented ovoid shape, high moisture (65.33-72.53%), low acidity, with succinic acid being the predominant organic acid, and high dietary fiber content (8.98-10.81%), mostly insoluble fibers (5.72-8.02%). Fruits from MP presented the highest amounts of free phenolic compounds (24.27 mg/100 g) and the highest antioxidant activity in the DPPH, FRAP and ORAC assays. In MP fruits, epigallocatechin gallate (12.04 mg/100 g) was the major compound, while in fruits from other states, phenolic compounds were the major compounds. The bioaccessibility of catechin, epicatechin, epicatechin gallate, epigallocatechin gallate, procyanidin and syringic acid varied among fruits from different regions. Fruits from SP presented the highest phenolic content in the bioaccessible fraction and highest antioxidant activity in all assays. The findings of this study indicate that juá is a non-acidic fruit with highest moisture, source of fiber, majority insoluble fibers and bioaccessible flavanols.
Assuntos
Catequina/análise , Fibras na Dieta/análise , Frutas/química , Extratos Vegetais/análise , Polifenóis/análise , Ziziphus/química , Antioxidantes/análise , Biflavonoides/análise , Brasil , Catequina/análogos & derivados , Fenômenos Químicos , Valor Nutritivo , Compostos Fitoquímicos/análise , Proantocianidinas/análiseRESUMO
INTRODUCTION: Introduction: casein-derived peptides can be liberated both in vivo via normal digestion of casein, as well as in vitro via enzymatic hydrolysis. These peptides were suggested to have biological activity. Objectives: the aim of this study was to describe the production and characterization of casein peptides and to explore the potential of these peptides as an option for low-phenylalanine diets. Methods: peptides were produced by tryptic hydrolysis of sodium caseinate and acid precipitation with HCl, followed by precipitation with ethanol or aggregation of CaCl2 or ZnSO4. Results: the amino acid analysis revealed a significant reduction in the amount of phenylalanine from the original protein. Conclusion: casein-derived peptides could be a future alternative of short chain peptides to low-phenylalanine formulations.
INTRODUCCIÓN: Introducción: los péptidos derivados de la caseína se pueden liberar tanto in vivo, a través de la digestión normal de la caseína, como in vitro a través de la hidrólisis enzimática. Se sugirió que estos péptidos tenían actividad biológica. Objetivos: el objetivo de este estudio fue describir la producción y caracterización de péptidos de caseína y explorar el potencial de estos péptidos como una opción para las dietas con bajo contenido de fenilalanina. Métodos: los péptidos se produjeron por hidrólisis tríptica de caseinato de sodio y precipitación ácida con HCl, seguido de precipitación con etanol o agregación de CaCl2 o ZnSO4. Resultados: el análisis de aminoácidos reveló una reducción significativa en la cantidad de fenilalanina de la proteína original. Conclusión: los péptidos derivados de la caseína podrían ser una alternativa futura de los péptidos de cadena corta a las formulaciones con bajo contenido de fenilalanina.
Assuntos
Caseínas/análise , Fenilalanina , Fenilcetonúrias/dietoterapia , Aminoácidos/análise , Aminoácidos/metabolismo , Caseínas/metabolismo , Dieta , Humanos , Peptídeos/análise , Peptídeos/metabolismoRESUMO
BACKGROUND: Impairments in cell migration during vertebrate gastrulation lead to structural birth defects, such as heart defects and neural tube defects. These defects are more frequent in progeny from diabetic pregnancies, and we have recently provided evidence that maternal diabetes leads to impaired migration of embryonic mesodermal cells in a mouse model of diabetic pregnancy. METHODS: We here report the isolation of primary cell lines from normal and diabetes-exposed embryos of the nonobese diabetic mouse strain, and characterization of their energy metabolism and expression of nutrient transporter genes by quantitative real-time PCR. RESULTS: Expression levels of several genes in the glucose transporter and fatty acid transporter gene families were altered in diabetes-exposed cells. Notably, primary cells from embryos with prior in vivo exposure to maternal diabetes exhibited reduced capacity for cell migration in vitro. CONCLUSIONS: Primary cells isolated from diabetes-exposed embryos retained a "memory" of their in vivo exposure, manifesting in cell migration impairment. Thus, we have successfully established an in vitro experimental model for the mesoderm migration defects observed in diabetes-exposed mouse embryos.
Assuntos
Movimento Celular/efeitos dos fármacos , Glucose/metabolismo , Gravidez em Diabéticas/fisiopatologia , Animais , Movimento Celular/fisiologia , Diabetes Mellitus Experimental , Diabetes Gestacional , Modelos Animais de Doenças , Embrião de Mamíferos/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Camundongos/embriologia , Camundongos Endogâmicos NOD , Defeitos do Tubo Neural/genética , Fator de Transcrição PAX3/genética , Gravidez , Fatores de Transcrição/genéticaRESUMO
Early podocyte loss is characteristic of chronic kidney diseases (CKD) in obesity and diabetes. Since treatments for hyperglycemia and hypertension do not prevent podocyte loss, there must be additional factors causing podocyte depletion. The role of oxidative stress has been implicated in CKD but it is not known how exactly free radicals affect podocyte physiology. To assess this relationship, we investigated the effects of lipid radicals on podocytes, as lipid peroxidation is a major form of oxidative stress in diabetes. We found that lipid radicals govern changes in podocyte homeostasis through redox sensitive RhoA signaling: lipid radicals inhibit migration and cause loss of F-actin fibers. These effects were prevented by mutating the redox sensitive cysteines of RhoA. We therefore suggest that in diseases associated with increased lipid peroxidation, lipid radicals can determine podocyte function with potentially pathogenic consequences for kidney physiology.
Assuntos
Peroxidação de Lipídeos/genética , Podócitos/metabolismo , Insuficiência Renal Crônica/genética , Proteína rhoA de Ligação ao GTP/genética , Actinas/genética , Actinas/metabolismo , Movimento Celular/genética , Citoesqueleto/genética , Citoesqueleto/metabolismo , Complicações do Diabetes/genética , Complicações do Diabetes/patologia , Radicais Livres/metabolismo , Humanos , Mutação , Obesidade/complicações , Obesidade/genética , Obesidade/patologia , Oxirredução , Podócitos/patologia , Insuficiência Renal Crônica/patologia , Transdução de SinaisRESUMO
Failure to close the neural tube results in birth defects, with severity ranging from spina bifida to lethal anencephaly. Few genetic risk factors for neural tube defects are known in humans, highlighting the critical role of environmental risk factors, such as maternal diabetes. Yet, it is not well understood how altered maternal metabolism interferes with embryonic development, and with neurulation in particular. We present evidence from two independent mouse models of diabetic pregnancy that identifies impaired migration of nascent mesodermal cells in the primitive streak as the morphogenetic basis underlying the pathogenesis of neural tube defects. We conclude that perturbed gastrulation not only explains the neurulation defects, but also provides a unifying etiology for the broad spectrum of congenital malformations in diabetic pregnancies.
Assuntos
Diabetes Gestacional/genética , Proteínas do Tecido Nervoso/genética , Placa Neural/metabolismo , Defeitos do Tubo Neural/genética , Animais , Diabetes Gestacional/metabolismo , Diabetes Gestacional/patologia , Modelos Animais de Doenças , Embrião de Mamíferos , Feminino , Gastrulação/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Microdissecção e Captura a Laser , Camundongos , Camundongos Endogâmicos NOD , Proteínas do Tecido Nervoso/metabolismo , Placa Neural/embriologia , Placa Neural/patologia , Defeitos do Tubo Neural/embriologia , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/patologia , GravidezRESUMO
Muitos métodos têm sido empregados na produção de peptídeos bioativos para a promoção da saúde. O objetivo deste estudo foi produzir caseinofosfopeptídeos por hidrólise tríptica do caseinato de sódio usando diferentes temperaturas (37 e 50 °C) e tempos de reação (2 e 4 h), caracterizá-los e analisar sua influência nas atividades citotóxica, antimicrobiana e antioxidante. Caseinofosfopeptídeos foram caracterizados através da composição centesimal, eletroforese em gel de poliacrilamida, Espectrometria de Massas e Cromatografia Líquida de Alta Eficiência. Toxicidade para leucócitos humanos, atividade antimicrobiana utilizando o teste de microdiluição em caldo e determinação da capacidade antioxidante pelo método de espécies reativas ao ácido tiobarbitúrico foram os ensaios biológicos realizados. Os resultados mostraram que as quatro frações peptídicas obtidas apresentaram-se com baixo peso molecular e elevados teores proteico e mineral; quanto ao perfil aminoacídico, apresentaram elevadas e diferenciadas quantidades de ácido glutâmico e serina, que pouco variaram de acordo com o processo de obtenção; não se mostraram tóxicos para leucócitos humanos; demonstraram atividade antimicrobiana para Escherichia coli e Salmonella Enteritidis e elevada capacidade antioxidante. Os resultados físico-químicos das frações de caseinofosfopeptídeos demonstraram elevada composição nutricional em termos de proteína e, principalmente, cálcio. O conjunto de dados indicou que alterações no tempo e na temperatura de reação para a obtenção dos hidrolisados não interferem nas suas qualidades biológicas, mostrando serem seguros para a promoção da saúde e para a aplicação em situações especiais, que envolvem pacientes desnutridos, imunossuprimidos, com comprometimento ósseo ou gastrintestinal decorrentes de inflamações e infecções.
Several methods have been employed for the production of bioactive peptides for health promotion. The aim of this study was to produce and characterize Casein phosphopeptides obtained by sodium caseinate tryptic hydrolysis under different temperatures (37 and 50 °C) and reaction times (2 and 4 h), and evaluate their biological capabilities. They have been characterized by assessing their centesimal composition, by polyacrylamide gel electrophoresis, Mass Spectrometry, and High-Performance Liquid Chromatography. The biological activities tested included toxicity for human leukocytes, antimicrobial assay using the microdilution test, and determination of the antioxidant capacity by the thiobarbituric acid reactive species method. The results showed that the four fractions obtained were of low molecular weight with high protein and mineral contents; their amino acid profile showed high and differentiated amounts of glutamic acid and serine independent of the methodological procedures. The results also showed no toxicity for human peripheral leukocytes, demonstrated antimicrobial activity against Escherichia coli and Salmonella Enteritidis as well as high antioxidant capacity. The results of the physicochemical Casein phosphopeptides' fractions showed high nutritional composition in terms of protein and, particularly, calcium. The biological assays indicated that time and temperature changes in the process for obtaining casein hydrolysates have not interfered with their biological qualities. In addition, they have proven safe in promoting health in special conditions involving malnourished and/or, immunocompromised patients or those with bone and/or gastrointestinal impairment due to inflammations and infections.
Assuntos
Anti-Infecciosos , Antioxidantes , Caseínas , FosfopeptídeosRESUMO
Structurally diverse bicyclo[3.3.0]octanes were prepared and tested for their biological activity. Both the antiproliferative activity and the results of phenotypic characterization varied with the substitution patterns. Two derivatives displayed high inhibitory (IC50 ≤3µM) activity against the L-929 cell line, but differed in their mode of action. A cluster analysis with impedance profiling data showed the two compounds in relationship to microtubule interfering compounds. In PtK2 cells treated with both derivatives a perturbing effect on the microtubular network was observed, whereas the actin cytoskeleton in incubated PtK2 cells was disturbed only by one compound. The effects on tubulin and actin polymerization could be confirmed by in vitro polymerization experiments.
Assuntos
Antineoplásicos/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Lactamas Macrocíclicas/farmacologia , Pirrolidinonas/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Humanos , Lactamas Macrocíclicas/síntese química , Lactamas Macrocíclicas/química , Camundongos , Conformação Molecular , Pirrolidinonas/síntese química , Pirrolidinonas/química , Relação Estrutura-AtividadeRESUMO
Albuminuria is associated with metabolic syndrome and diabetes. It correlates with the progression of chronic kidney disease, particularly with tubular atrophy. The fatty acid load on albumin significantly increases in obesity, presenting a proinflammatory environment to the proximal tubules. However, little is known about changes in the redox milieu during fatty acid overload and how redox-sensitive mechanisms mediate cell death. Here, we show that albumin with fatty acid impurities or conjugated with palmitate but not albumin itself compromised mitochondrial and cell viability, membrane potential and respiration. Fatty acid overload led to a redox imbalance which deactivated the antioxidant protein peroxiredoxin 2 and caused a peroxide-mediated apoptosis through the redox-sensitive pJNK/caspase-3 pathway. Transfection of tubular cells with peroxiredoxin 2 was protective and mitigated apoptosis. Mitochondrial fatty acid entry and ceramide synthesis modulators suggested that mitochondrial ß oxidation but not ceramide synthesis may modulate lipotoxic effects on tubular cell survival. These results suggest that albumin overloaded with fatty acids but not albumin itself changes the redox environment in the tubules, inducing a peroxide-mediated redox-sensitive apoptosis. Thus, mitigating circulating fatty acid levels may be an important factor in both preserving redox balance and preventing tubular cell damage in proteinuric diseases.
Assuntos
Albuminas/metabolismo , Apoptose/efeitos dos fármacos , Ácidos Graxos/farmacologia , Albuminas/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Células Epiteliais/patologia , Túbulos Renais Proximais/citologia , Túbulos Renais Proximais/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/metabolismo , Oxirredução , Palmitatos/metabolismo , Peroxirredoxinas/metabolismo , Ratos , Soroalbumina Bovina/farmacologiaRESUMO
Periconceptional supplementation of folic acid to the diet of women is considered a great success for a public health intervention. Higher folate status, either by supplementation, or via the mandatory fortification of grain products in the United States, has led to significant reduction in the incidence of neural tube defects. Besides birth defects, folate deficiency has been linked to a variety of morbidities, most notably to increased risk for cancer. However, recent evidence suggests that excess folate may be detrimental - for birth defect incidence or in the progression of cancer. How folate mediates beneficial or detrimental effects is not well understood. It is also unknown what molecular responses are elicited in women taking folate supplements, and thus experience a bolus of folate on top of the status achieved by fortification. To characterize the response to a periconceptional regimen of supplementation with folinic acid, we performed gene expression profiling experiments on uterus tissue of pregnant mice with either wildtype alleles or targeted disruption at the folate receptor 4 locus. We observed that, depending on the genetic background, folinic acid supplementation affects expression of genes that contribute to lipid metabolism, protein synthesis, mitochondrial function, cell cycle, and cell activation. The extent of the response is strongly modulated by the genetic background. Finally, we provide evidence that folinic acid supplementation in the mutant paradigm affects histone methylation status, a potential mechanism of gene regulation in this model.
Assuntos
Ácido Fólico/administração & dosagem , Perfilação da Expressão Gênica , Mutação , Receptores de Superfície Celular/genética , Útero/metabolismo , Animais , Feminino , Genótipo , Camundongos , Camundongos Endogâmicos C57BL , GravidezRESUMO
BACKGROUND: How signals from fatty acid metabolism are translated into changes in food intake remains unclear. Previously we reported that mice with a genetic inactivation of Acads (acyl-coenzyme A dehydrogenase, short-chain), the enzyme responsible for mitochondrial beta-oxidation of C4-C6 short-chain fatty acids (SCFAs), shift consumption away from fat and toward carbohydrate when offered a choice between diets. In the current study, we sought to indentify candidate genes and pathways underlying the effects of SCFA oxidation deficiency on food intake in Acads-/- mice. METHODOLOGY/PRINCIPAL FINDINGS: We performed a transcriptional analysis of gene expression in brain tissue of Acads-/- and Acads+/+ mice fed either a high-fat (HF) or low-fat (LF) diet for 2 d. Ingenuity Pathway Analysis revealed three top-scoring pathways significantly modified by genotype or diet: oxidative phosphorylation, mitochondrial dysfunction, and CREB signaling in neurons. A comparison of statistically significant responses in HF Acads-/- vs. HF Acads+/+ (3917) and Acads+/+ HF vs. LF Acads+/+ (3879) revealed 2551 genes or approximately 65% in common between the two experimental comparisons. All but one of these genes were expressed in opposite direction with similar magnitude, demonstrating that HF-fed Acads-deficient mice display transcriptional responses that strongly resemble those of Acads+/+ mice fed LF diet. Intriguingly, genes involved in both AMP-kinase regulation and the neural control of food intake followed this pattern. Quantitative RT-PCR in hypothalamus confirmed the dysregulation of genes in these pathways. Western blotting showed an increase in hypothalamic AMP-kinase in Acads-/- mice and HF diet increased, a key protein in an energy-sensing cascade that responds to depletion of ATP. CONCLUSIONS: Our results suggest that the decreased beta-oxidation of short-chain fatty acids in Acads-deficient mice fed HF diet produces a state of energy deficiency in the brain and that AMP-kinase may be the cellular energy-sensing mechanism linking fatty acid oxidation to feeding behavior in this model.
Assuntos
Acil-CoA Desidrogenase/deficiência , Dieta Hiperlipídica/efeitos adversos , Transdução de Sinais/efeitos dos fármacos , Acil-CoA Desidrogenase/genética , Trifosfato de Adenosina/metabolismo , Animais , Encéfalo , Ácidos Graxos/metabolismo , Hipotálamo/metabolismo , Masculino , Camundongos , Camundongos Mutantes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/genéticaRESUMO
Pregnancies complicated by maternal diabetes have long been known to carry a higher risk for congenital malformations, such as neural tube defects. Using the FVB inbred mouse strain and the Streptozotocin-induced diabetes model, we tested whether the incidence of neural tube defects in diabetic pregnancies can be modulated by maternal diet. In a comparison of two commercial mouse diets, which are considered nutritionally replete, we found that maternal consumption of the unfavorable diet was associated with a more than 3-fold higher rate of neural tube defects. Our results demonstrate that maternal diet can act as a modifier of the risk for abnormal development in high-risk pregnancies, and provide support for the possibility that neural tube defects in human diabetic pregnancies might be preventable by optimized maternal nutrition.
Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Diabetes Mellitus Experimental , Fenômenos Fisiológicos da Nutrição Materna , Defeitos do Tubo Neural/prevenção & controle , Gravidez em Diabéticas , Ração Animal , Animais , Modificador do Efeito Epidemiológico , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário , Feminino , Camundongos , Defeitos do Tubo Neural/embriologia , Defeitos do Tubo Neural/epidemiologia , Gravidez , RiscoRESUMO
The transport of metal micronutrients to developing organs in a plant is mediated primarily by the sieve elements. Ligands are thought to form complexes with the free ions in order to prevent cellular damage, but no binding partners have been unequivocally identified from plants so far. This study has used the phloem-mediated transport of micronutrients during the germination of the castor bean seedling to identify an iron transport protein (ITP). It is demonstrated that essentially all (55)Fe fed to seedlings is associated with the protein fraction of phloem exudate. It is shown that ITP carries iron in vivo and binds additional iron in vitro. ITP was purified to homogeneity from minute amounts of phloem exudate using immobilized metal ion affinity chromatography. It preferentially binds to Fe(3+) but not to Fe(2+) and also complexes Cu(2+), Zn(2+), and Mn(2+) in vitro. The corresponding cDNA of ITP was cloned using internal peptide fragments. The deduced protein of 96 amino acids shows high similarity to the stress-related family of late embryogenesis abundant proteins. Its predicted characteristics and its RNA expression pattern are consistent with a function in metal ion binding. The ITP from Ricinus provides the first identified micronutrient binding partner for phloem-mediated long distance transport in plants and is the first member of the late embryogenesis abundant protein family shown to have such a function.