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1.
Theor Appl Genet ; 128(7): 1431-47, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25930056

RESUMO

KEY MESSAGE: The identification of genetic factors influencing the accumulation of individual glucosinolates in broccoli florets provides novel insight into the regulation of glucosinolate levels in Brassica vegetables and will accelerate the development of vegetables with glucosinolate profiles tailored to promote human health. Quantitative trait loci analysis of glucosinolate (GSL) variability was conducted with a B. oleracea (broccoli) mapping population, saturated with single nucleotide polymorphism markers from a high-density array designed for rapeseed (Brassica napus). In 4 years of analysis, 14 QTLs were associated with the accumulation of aliphatic, indolic, or aromatic GSLs in floret tissue. The accumulation of 3-carbon aliphatic GSLs (2-propenyl and 3-methylsulfinylpropyl) was primarily associated with a single QTL on C05, but common regulation of 4-carbon aliphatic GSLs was not observed. A single locus on C09, associated with up to 40 % of the phenotypic variability of 2-hydroxy-3-butenyl GSL over multiple years, was not associated with the variability of precursor compounds. Similarly, QTLs on C02, C04, and C09 were associated with 4-methylsulfinylbutyl GSL concentration over multiple years but were not significantly associated with downstream compounds. Genome-specific SNP markers were used to identify candidate genes that co-localized to marker intervals and previously sequenced Brassica oleracea BAC clones containing known GSL genes (GSL-ALK, GSL-PRO, and GSL-ELONG) were aligned to the genomic sequence, providing support that at least three of our 14 QTLs likely correspond to previously identified GSL loci. The results demonstrate that previously identified loci do not fully explain GSL variation in broccoli. The identification of additional genetic factors influencing the accumulation of GSL in broccoli florets provides novel insight into the regulation of GSL levels in Brassicaceae and will accelerate development of vegetables with modified or enhanced GSL profiles.


Assuntos
Brassica/química , Brassica/genética , Glucosinolatos/química , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Mapeamento Cromossômico , Cromossomos de Plantas , DNA de Plantas/genética , Flores/química , Flores/genética , Ligação Genética , Marcadores Genéticos , Fenótipo , Verduras/química , Verduras/genética
2.
Int J Food Sci Nutr ; 66(2): 175-85, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25519247

RESUMO

Cost-effective methods for concentration and stabilization of otherwise perishable mango fruit phytoactives into shelf stable high protein ingredients were developed to combat stunting (malnutrition) in rural Africa. Mango juices complexed with sunflower oil and protein-rich legume flours yielded carotenoid-enriched oils and pelleted polyphenol-enriched flour matrices. Carotenoids from juices were concentrated 9-10 times in the fortified sunflower oil. Protein-rich soy and peanut flours captured 2.2-3.2 mg/g polyphenols from the juices. Alternatively, mango juice was sorbed and co-dried with flours, which stably bound the polyphenols, carotenoids, and natural sugars in soy or peanut protein-rich matrices. The concentration of provitamin A carotenoids was almost doubled and total polyphenols were enriched 4-5 times higher in the matrices compared to fresh pureed juice. Both strategies require minimal instrumentation, are compatible with rural village dietary practices; and capture the benefits of otherwise perishable seasonal resources by complexing healthful proteins together with phytoactive compounds.


Assuntos
Arachis , Carotenoides/análise , Proteínas Alimentares , Glycine max , Helianthus , Mangifera/química , Polifenóis/análise , África , Carboidratos/análise , Carotenoides/administração & dosagem , Dieta , Farinha/análise , Manipulação de Alimentos/métodos , Conservação de Alimentos , Frutas/química , Saúde , Humanos , Desnutrição/prevenção & controle , Compostos Fitoquímicos/análise , Óleos de Plantas , Polifenóis/administração & dosagem , População Rural , Óleo de Girassol
3.
Theor Appl Genet ; 127(9): 2051-64, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25119868

RESUMO

KEY MESSAGE: A high-resolution genetic linkage map of B. oleracea was developed from a B. napus SNP array. The work will facilitate genetic and evolutionary studies in Brassicaceae. A broccoli population, VI-158 × BNC, consisting of 150 F2:3 families was used to create a saturated Brassica oleracea (diploid: CC) linkage map using a recently developed rapeseed (Brassica napus) (tetraploid: AACC) Illumina Infinium single nucleotide polymorphism (SNP) array. The map consisted of 547 non-redundant SNP markers spanning 948.1 cM across nine chromosomes with an average interval size of 1.7 cM. As the SNPs are anchored to the genomic reference sequence of the rapid cycling B. oleracea TO1000, we were able to estimate that the map provides 96 % coverage of the diploid genome. Carotenoid analysis of 2 years data identified 3 QTLs on two chromosomes that are associated with up to half of the phenotypic variation associated with the accumulation of total or individual compounds. By searching the genome sequences of the two related diploid species (B. oleracea and B. rapa), we further identified putative carotenoid candidate genes in the region of these QTLs. This is the first description of the use of a B. napus SNP array to rapidly construct high-density genetic linkage maps of one of the constituent diploid species. The unambiguous nature of these markers with regard to genomic sequences provides evidence to the nature of genes underlying the QTL, and demonstrates the value and impact this resource will have on Brassica research.


Assuntos
Brassica/genética , Mapeamento Cromossômico , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Carotenoides/genética , DNA de Plantas/genética , Ligação Genética , Genoma de Planta
4.
J Sci Food Agric ; 94(13): 2639-48, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24497378

RESUMO

BACKGROUND: Maqui (Aristotelia chilensis) is a Chilean species which produces small berries that are collected from the wild. Anthocyanins, because of their health benefits, are the major focus of interest in maqui fruit. For this study, we examined anthocyanin and phenolic content of maqui fruits from individuals that belonged to four geographical areas in Chile, and used DNA marker analysis to examine the genetic variability of maqui populations that had distinctly different fruit anthocyanin content. RESULTS: Twelve primers generated a total of 145 polymorphic inter simple sequence repeat-polymerase chain reaction (ISSR-PCR) bands. ISSR-PCR showed different banding patterns for the individuals evaluated, confirming that maqui populations belonged to different genotypes. Maqui fruit from four different geographical regions during two consecutive growing seasons showed high total anthocyanin (6.6-15.0 g cy-3-glu kg⁻¹ fresh weight (FW)) and phenolic (10.7-20.5 g GAE kg⁻¹ FW) contents and different anthocyanin profiles. CONCLUSION: Three maqui genotypes exhibited significantly higher anthocyanin content than the others, as measured by pH differential method and high-performance liquid chromatography-mass spectrometry. Significant genetic diversity was noted within each ecological population. ISSR-PCR analysis provided a fingerprinting approach applicable for differentiation of maqui genotypes.


Assuntos
Antocianinas/análise , Antioxidantes/análise , Elaeocarpaceae/química , Frutas/química , Altitude , Antocianinas/metabolismo , Antioxidantes/metabolismo , Chile , Cromatografia Líquida de Alta Pressão , Clima , Elaeocarpaceae/genética , Elaeocarpaceae/crescimento & desenvolvimento , Elaeocarpaceae/metabolismo , Frutas/genética , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Marcadores Genéticos , Variação Genética , Limite de Detecção , Fenóis/análise , Fenóis/metabolismo , Filogenia , Análise Espaço-Temporal , Especificidade da Espécie , Espectrometria de Massas por Ionização por Electrospray , Meio Selvagem
5.
Plant Foods Hum Nutr ; 69(4): 372-8, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25359555

RESUMO

Well-known health-protective phytochemicals from muscadine grape and kale were stably complexed with food grade protein (soy or hemp protein isolates) to create biofortified food ingredients for use in a variety of convenient, portable food formulations. The bioactive (anti-inflammatory) potential, sensory attributes and proximates of the prepared formulations were evaluated in this study. Anti-inflammatory properties of the protein-phytoactive ingredient particles were contributed by the polyphenolic content (muscadine-protein) or the combination of polyphenol, carotenoid, and glucosinolate content (kale-protein aggregates). Phytoactive compounds from the fortified matrices suppressed at least two biomarkers of inflammation; most notable with the expression of chronic pro-inflammatory genes IL-6 and Mcp1. Sensory analysis suggested both sweet and savory functional food applications for the biofortified ingredients. Proximate analyses determined that fortification of the soy protein isolate (SPI) with muscadine or kale bioactives resulted in elevated dietary fibers, total carbohydrates, and free sugars, but did not increase calories/100 g dry matrix compared to unfortified SPI. Overall protein content in the aggregate matrices was about 37% less (muscadine-SPI, kale-SPI and kale- HP50) or 17.6% less (muscadine-HP50) on a weight basis, likely due to solubility of some proteins during preparation and partial displacement of some protein mass by the fruit and vegetable phytoactive constituents.


Assuntos
Anti-Inflamatórios/uso terapêutico , Brassica/química , Dieta , Proteínas Alimentares/análise , Alimento Funcional/análise , Paladar , Vitis/química , Animais , Anti-Inflamatórios/farmacologia , Cannabis , Carotenoides/farmacologia , Carotenoides/uso terapêutico , Quimiocina CCL2/metabolismo , Frutas/química , Glucosinolatos/farmacocinética , Glucosinolatos/farmacologia , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Interleucina-6/metabolismo , Camundongos , Valor Nutritivo , Preparações de Plantas/farmacologia , Preparações de Plantas/uso terapêutico , Polifenóis/farmacologia , Polifenóis/uso terapêutico , Proteínas de Soja , Verduras/química
6.
Plant Foods Hum Nutr ; 69(4): 317-24, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25351938

RESUMO

Co-delivery of edible proteins with health-protective fruit (muscadine grape) and vegetable (kale) phytoactive compounds was accomplished in a biofortified ingredient for use in convenient, portable food formulations. Polyphenolics were concentrated (10-42 mg/g range) in dry muscadine-protein matrices. Kale-fortified protein matrices also captured polyphenolics (8 mg/g), carotenoids (69 µg/g) and glucosinolates (7 µmol/g). Neither total phenolics nor glucosinolates were significantly diminished even after long term (6 months) storage at 4, 20, or 37 °C, whereas carotenoids degraded over time, particularly at higher temperatures. Dry biofortified phytoactive-protein ingredients allowed delivery of immunoprotective compounds from fruits and vegetables in a stable, lightweight matrix.


Assuntos
Brassica/química , Carotenoides/análise , Proteínas Alimentares , Alimento Funcional , Glucosinolatos/análise , Polifenóis/análise , Vitis/química , Dieta , Manipulação de Alimentos , Frutas/química , Humanos , Fatores Imunológicos , Extratos Vegetais/química , Verduras/química
7.
bioRxiv ; 2023 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-37163110

RESUMO

Parkinson's disease (PD) is a neurodegenerative disorder involving motor symptoms caused by a loss of dopaminergic neurons in the substantia nigra region of the brain. Epidemiological evidence suggests that anthocyanin (ANC) intake is associated with a low risk of PD. Previously, we reported that extracts enriched with ANC and proanthocyanidins (PAC) suppressed dopaminergic neuron death elicited by the PD-related toxin rotenone in a primary midbrain culture model. Here, we characterized botanical extracts enriched with a mixed profile of polyphenols, as well as a set of purified polyphenolic standards, in terms of their ability to mitigate dopaminergic cell death in midbrain cultures exposed to another PD-related toxicant, paraquat (PQ), and we examined underlying neuroprotective mechanisms. Extracts prepared from blueberries, black currants, grape seeds, grape skin, mulberries, and plums, as well as several ANC, were found to rescue dopaminergic neuron loss in PQ-treated cultures. Comparison of a subset of ANC-rich extracts for the ability to mitigate neurotoxicity elicited by PQ versus rotenone revealed that a hibiscus or plum extract was only neuroprotective in cultures exposed to rotenone or PQ, respectively. Several extracts or compounds with the ability to protect against PQ neurotoxicity increased the activity of the antioxidant transcription factor Nrf2 in cultured astrocytes, and PQ-induced dopaminergic cell death was attenuated in Nrf2-expressing midbrain cultures. In other studies, we found that extracts prepared from hibiscus, grape skin, or purple basil (but not plums) rescued defects in O 2 consumption in neuronal cells treated with rotenone. Collectively, these findings suggest that extracts enriched with certain combinations of ANC, PAC, stilbenes, and other polyphenols could potentially slow neurodegeneration in the brains of individuals exposed to PQ or rotenone by activating cellular antioxidant mechanisms and/or alleviating mitochondrial dysfunction.

8.
Antioxidants (Basel) ; 12(7)2023 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-37508001

RESUMO

Parkinson's disease (PD) is a neurodegenerative disorder involving motor symptoms caused by a loss of dopaminergic neurons in the substantia nigra region of the brain. Epidemiological evidence suggests that anthocyanin (ANC) intake is associated with a low risk of PD. Previously, we reported that extracts enriched with ANC and proanthocyanidins (PAC) suppressed dopaminergic neuron death elicited by the PD-related toxin rotenone in a primary midbrain culture model. Here, we characterized botanical extracts enriched with a mixed profile of polyphenols, as well as a set of purified polyphenolic standards, in terms of their ability to mitigate dopaminergic cell death in midbrain cultures exposed to another PD-related toxicant, paraquat (PQ), and we examined underlying neuroprotective mechanisms. Extracts prepared from blueberries, black currants, grape seeds, grape skin, mulberries, and plums, as well as several ANC, were found to rescue dopaminergic neuron loss in PQ-treated cultures. Comparison of a subset of ANC-rich extracts for the ability to mitigate neurotoxicity elicited by PQ versus rotenone revealed that a hibiscus or plum extract was only neuroprotective in cultures exposed to rotenone or PQ, respectively. Several extracts or compounds with the ability to protect against PQ neurotoxicity increased the activity of the antioxidant transcription factor Nrf2 in cultured astrocytes, and PQ-induced dopaminergic cell death was attenuated in Nrf2-expressing midbrain cultures. In other studies, we found that extracts prepared from hibiscus, grape skin, or purple basil (but not plums) rescued defects in O2 consumption in neuronal cells treated with rotenone. Collectively, these findings suggest that extracts enriched with certain combinations of ANC, PAC, stilbenes, and other polyphenols could potentially slow neurodegeneration in the brains of individuals exposed to PQ or rotenone by activating cellular antioxidant mechanisms and/or alleviating mitochondrial dysfunction.

9.
J Sep Sci ; 35(13): 1682-9, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22761148

RESUMO

Blackcurrant is considered as a natural high-value food raw material and possesses a variety of therapeutic properties. The health benefits of blackcurrant have generally been credited to its high anthocyanin content; however, the therapeutic properties of other minor flavonoids constituents have not yet been investigated due the difficulties related to their isolation. Multiple steps of high-performance counter-current chromatography in combination with ESI tandem mass spectrometry (MS(n)) were successfully used for the preparative isolation of flavonols from blackcurrant extract, to study their electrospray ionization mass spectrometry fragmentation behavior. Seven flavonols, namely myricetin-3-O-rutinoside (145.5 mg), myricetin-3-O-hexoside (79.7 mg), myricetin-3-O-(6″-malonyl)-glucoside (17.4 mg), kaempferol-3-O-glucoside (20.5 mg), quercetin-3-O-rutinoside (55.1 mg), quercetin-3-O-hexoside (25.8 mg), and myricetin (129.1 mg) have been successfully isolated and their multistage MS(n) data were used for detailed structure characterization. The results of these experiments demonstrated that high-performance counter-current chromatography along with ESI-MS(n) is a sensitive, selective, and effective technology for isolation and characterization of minor constituents from a complex mixture.


Assuntos
Distribuição Contracorrente/métodos , Flavonóis/química , Flavonóis/isolamento & purificação , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Ribes/química , Espectrometria de Massas por Ionização por Electrospray/métodos
10.
Nucl Instrum Methods Phys Res B ; 268(7-8): 1313-1316, 2010 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-20419067

RESUMO

Bioactive compounds in botanicals may be beneficial in preventing age-related neurodegenerative diseases, but for many compounds conventional methods may be inadequate to detect if these compounds cross the blood brain barrier or to track the pharmacokinetics in the brain. By combining a number of unique technologies it has been possible to utilize the power of AMS to study the pharmacokinetics of bioactive compounds in the brain at very low concentrations. (14)C-labeled compounds can be biosynthesized by plant cell suspension cultures co-incubated with radioisotopically-labeled sucrose and isolated and separated into a series of bioactive fractions.To study the pharmacokinetics and tissue distribution of (14)C labeled plant polyphenols, rats were implanted with jugular catheters, subcutaneous ultrafiltration probes and brain microdialysis probes. Labeled fractions were dosed orally. Interstitial fluid (ISF) and brain microdialysate samples were taken in tandem with blood samples. It was often possible to determine (14)C in blood and ISF with a ß-counter. However, brain microdialysate samples (14)C levels on the order of 10(7) atoms/sample required AMS technology. The Brain Microdialysate(AUC)/Serum(AUC) ranged from .021- to .029, with the higher values for the glycoside fractions. By using AMS in combination with traditional methods, it is possible to study uptake by blood, distribution to ISF and determine the amount of a dose which can reach the brain and follow the pharmacokinetics in the brain.

11.
Br J Nutr ; 102(10): 1523-30, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19586574

RESUMO

Absorption, distribution and elimination of 14C-labelled isoflavone-containing extracts from kudzu (Pueraria lobata) root culture and red clover (Trifolium pratense) cell culture were investigated in an in vivo rat model. The predominant isoflavones in the kudzu extract were the glycosides puerarin, daidzin and malonyl daidzin, while in the red clover extract, the major isoflavones were formononetin and its derivatives, genistein and biochanin A, with radioactivities of 3.770 and 7.256 MBq/g, respectively. Male Sprague-Dawley rats, implanted with a jugular catheter and a subcutaneous ultrafiltrate probe, were orally administered with 14C-labelled isoflavone extracts from either kudzu or clover cell cultures. Serum, interstitial fluid (ISF), urine and faeces were collected using a Culex Automated Blood Collection System for 24 h. Analysis of bone tissues revealed that radiolabel accumulated in the femur, tibia and vertebrae at 0.04, 0.03 and 0.01 % of the administered dose, respectively, in both kudzu and red clover treatments. The liver accumulated the greatest concentration of radiolabel among the tissues tested, at 1.99 and 1.54 % of the administered kudzu and red clover extracts, respectively. Serum and ISF analysis showed that both extracts were rapidly absorbed, distributed in various tissues, and largely eliminated in the urine and faeces. Urine and faeces contained 8.53 and 9.06 % of the kudzu dose, respectively, and 3.60 and 5.64 % of the red clover dose, respectively. Serum pharmacokinetics suggest that extracts from kudzu may undergo enterohepatic circulation.


Assuntos
Carbono/química , Isoflavonas/metabolismo , Pueraria/química , Trifolium/química , Animais , Radioisótopos de Carbono , Células Cultivadas , Líquido Extracelular/química , Isoflavonas/sangue , Isoflavonas/química , Masculino , Raízes de Plantas/citologia , Ratos , Ratos Sprague-Dawley , Fatores de Tempo
12.
Food Chem ; 301: 125289, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31387047

RESUMO

Acylated anthocyanins, such as those found in red cabbage, are more heat-, light-, and alkaline pH-stable than non-acylated anthocyanins, making them attractive for a variety of commercial applications. A UPLC-DAD-MSE method with an optimized chromatographic strategy was used to identify 29 red cabbage anthocyanins, predominantly acylated and glucosylated cyanidin derivatives. Anthocyanin profiles of 27 red cabbage genotypes harvested in consecutive growing seasons were measured and assessed for variation. Three unique anthocyanin profile fingerprints were identified through hierarchical clustering analysis. PCA analysis identified anthocyanin accumulation traits and genotypes with high diversity which can be utilized in future investigations into the genetic and molecular basis for anthocyanin production, acylation, and diversity.


Assuntos
Antocianinas/análise , Brassica/química , Brassica/genética , Melhoramento Vegetal , Polimorfismo Genético , Estações do Ano , Acilação , Antocianinas/química , Brassica/metabolismo , Cromatografia Líquida de Alta Pressão , Genótipo , Espectrometria de Massas
13.
J Agric Food Chem ; 56(6): 1896-906, 2008 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-18284202

RESUMO

Extracts rich in proanthocyanidins, which are implicated in multiple human health benefits, were comparatively separated using alternative separation methods [vacuum or open column liquid chromatography], separation matrices [Toyopearl, Sephadex, or silica gel], and degrees of subfractionation [8 or 12 subfraction series], to evaluate the influence of separation technique on the resolution of the chemical composition and the biological activity of separated proanthocyanidin mixtures in individual subfractions. Bioactivity was assessed using a DNA human topoisomerase II bioassay and structural composition by acid thiolysis (average degree of polymerization, DP) and HPLC-ESI/MS. The amount of parent fraction needed to inhibit 50% of topoisomerase II was 3.38 ng/mL with an average DP of 25.5. A 2(3) factorial analysis revealed that the vacuum and open column strategies for separation, when individually considered, did not yield significantly different results in terms of mass recovery, DP, or bioactivity; however, interactions with other factors such as matrix or subfraction series resulted in distinctive shifts in fraction profiles and biological activity. In general, Sephadex as a matrix permitted elution and separation of discrete, polymerized subfractions with potent inhibition against human topoisomerase II. Sephadex vacuum chromatography, Toyopearl open column chromatography, and Toyopearl vacuum chromatography separation techniques eluted highly polymerized proanthocyanidin mixtures, but the inhibitory bioactivity was attenuated as compared to the parent fraction, whereas Sephadex open column chromatography eluted highly polymerized subfraction mixtures that retained bioactive potential.


Assuntos
Cromatografia Líquida/métodos , Extratos Vegetais/química , Proantocianidinas/química , Cromatografia Líquida de Alta Pressão , Inibidores Enzimáticos/farmacologia , Humanos , Extratos Vegetais/farmacologia , Proantocianidinas/isolamento & purificação , Espectrometria de Massas por Ionização por Electrospray , Inibidores da Topoisomerase II
14.
Phytochemistry ; 67(21): 2380-91, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16956631

RESUMO

In comparison to the well-recognized adaptogenic herb Rhodiola rosea, phytochemical constituents of two other Rhodiola species (R. heterodonta and R. semenovii) were elucidated and characterized. Two major phytochemical groups; phenolic and/or cyanogenic glycosides and proanthocyanidins, were isolated and identified in the three species. Chemical similarities among the three species were observed; however, each species displayed differences in phytochemical constituents. R. heterodonta contained a newly detected phenylethanoid glycoside, heterodontoside, in addition to the known compounds tyrosol, viridoside, salidroside, and rhodiocyanoside A. Both R. heterodonta and R. rosea contained phenylethanoid/propanoid compounds that were not detected in R. semenovii. For R. semenovii, the cyanogenic glucosides rhodiocyanoside A and lotaustralin were detected. Although the three species have proanthocyanidins composed of (-)-epigallocatechin and its 3-O-gallate esters in common, the degree of polymerization greatly differed between them. In contrast to R. heterodonta and R. semenovii, R. rosea has higher molecular weight polymeric proanthocyanidins. This study resulted in the identification and isolation of phytochemical constituents for direct cross-comparison between three Rhodiola species of medicinal and pharmacological value.


Assuntos
Rhodiola/química , Rhodiola/metabolismo , Glucosídeos/química , Glicosídeos/química , Estrutura Molecular , Fenóis/química , Proantocianidinas/química , Rhodiola/classificação , Especificidade da Espécie
15.
J Agric Food Chem ; 54(6): 2116-22, 2006 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-16536584

RESUMO

The antiproliferative effects of 100% methanol crude extract and of Toyopearl and silica gel fractions from the seed coats of black Jamapa beans (Phaseolus vulgaris L.) were evaluated using HeLa, human adenocarcinoma cells, and HaCaT, human premalignant keratinocytes. The 100% methanol crude extract [172.2 microM equiv of (+)-catechin] increased adhesion of HeLa cells; however, 3- and 5-fold higher concentrations decreased the number of cells attached as a function of the treatment time. The highest concentration tested diminished the cell adhesion until 40% (after 24 h) to almost 80% (after 72 h). The IC50 values showed that the 100% methanol crude extract was the most effective inhibitor of HeLa cell proliferation, even when it was dissolved in dimethylsulfoxide (DMSO) [34.5 microM equiv of (+)-catechin] or in medium [97.7 microM equiv of (+)-catechin]. The Toyopearl 5 (TP5) fraction and silica gel 2 (SG2) fraction inhibited 60% of the HeLa cell proliferation. The IC50 was 154 microM equiv of (+)-catechin of the 100% methanol crude extract on HaCaT cells. Toyopearl fractions TP4 and TP6 significantly inhibited HaCaT cell proliferation, but the silica gel fractions did not have a significant effect. The 100% methanol crude extract (35 microg of dry material/mL) decreased the number of HeLa cells in the G0/G1 phase from 68.9% (for control cells) to 51.4% (for treated cells) and increased apoptosis (2.9 and 21.2% for control and treated cells, respectively). The results indicated that black Jamapa beans could be a source of polyphenolic compounds, which have an inhibitory effect toward HeLa cancer cells but are less aggressive on HaCaT premalignant cells.


Assuntos
Anticarcinógenos/farmacologia , Flavonoides/farmacologia , Phaseolus/química , Fenóis/farmacologia , Sementes/química , Apoptose/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Células HeLa , Humanos , Queratinócitos , Metanol , Extratos Vegetais/farmacologia , Polifenóis , Lesões Pré-Cancerosas , Neoplasias Cutâneas
16.
J Agric Food Chem ; 53(11): 4615-22, 2005 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-15913334

RESUMO

The common bean contains phytochemicals, including phenolic compounds, which can provide health benefits to the consumer. Our objective was to characterize the polyphenolic compounds present in the seed coat of Black Jamapa bean and to test fractionation methods that permit the recovery of polyphenolics in their naturally occurring forms. A 100% methanol extract from seed coats was subjected to different chromatographic fractionation methods. Analysis by HPLC-MS revealed that a better separation of phytochemicals was achieved using direct silica gel fractionation, which allowed more accurate identification of compounds, especially of the flavonols. Anthocyanins, flavanol monomers, and heterogeneous flavanol oligomers up to hexamers were detected. To our knowledge, this is the first time that myricetin glycoside and proanthocyanidin oligomers containing (epi)-gallocatechin have been reported in the black bean. The fractionation methods used in this study produced large quantities of natural mixtures of flavonoids suitable for testing bioactivity and phytochemical interactions.


Assuntos
Flavonoides/análise , Phaseolus/química , Fenóis/análise , Sementes/química , Antocianinas/análise , Fracionamento Químico , Cromatografia Líquida de Alta Pressão , Flavonoides/isolamento & purificação , Espectrometria de Massas , Metanol , Fenóis/isolamento & purificação , Extratos Vegetais/química , Polifenóis , Proantocianidinas/análise
17.
Artigo em Inglês | MEDLINE | ID: mdl-26255696

RESUMO

A high-throughput, robust and reliable method for simultaneous analysis of five carotenoids, four chlorophylls and one tocopherol was developed for rapid screening large sample populations to facilitate molecular biology and plant breeding. Separation was achieved for 10 known analytes and four unknown carotenoids in a significantly reduced run time of 10min. Identity of the 10 analytes was confirmed by their UV-Vis absorption spectras. Quantification of tocopherol, carotenoids and chlorophylls was performed at 290nm, 460nm and 650nm respectively. In this report, two sub two micron particle core-shell columns, Kinetex from Phenomenex (1.7µm particle size, 12% carbon load) and Cortecs from Waters (1.6µm particle size, 6.6% carbon load) were investigated and their separation efficiencies were evaluated. The peak resolutions were >1.5 for all analytes except for chlorophyll-a' with Cortecs column. The ruggedness of this method was evaluated in two identical but separate instruments that produced CV<2 in peak retentions for nine out of 10 analytes separated.


Assuntos
Carotenoides/análise , Clorofila/análise , Tocoferóis/análise , Calibragem , Ensaios de Triagem em Larga Escala , Limite de Detecção , Reprodutibilidade dos Testes , Espectrofotometria Ultravioleta
18.
J Agric Food Chem ; 52(5): 1138-45, 2004 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-14995111

RESUMO

A range of radiolabeled anthocyanins, proanthocyanidins, and other flavonoids were accumulated by cell suspension cultures of two plant species, ohelo (Vaccinium pahalae) and grape (a Vitis hybrid, Bailey Alicant A), after providing uniformly labeled [(14)C]sucrose to the medium. Approximately 15% of administered label was recovered in a series of flavonoid-rich fractions varying in composition. Anthocyanins, and monomers to oligomers of proanthocyanidins, were labeled effectively and characterized from both species. Most of the proanthocyanidin oligomers were based on the flavan-3-ols (+)-catechin and (-)-epicatechin. Cyanidin and peonidin glycosides were the dominant forms of anthocyanins in both species. Whereas the predominant form of flavonoids identified from ohelo cell cultures was proanthocyanidins, grape cell cultures produced mostly anthocyanins. The labeled phytochemicals were produced for use in subsequent in vivo animal feeding studies to gauge their bioavailability and accumulation in target organs.


Assuntos
Flavonoides/biossíntese , Vaccinium/metabolismo , Vitis/metabolismo , Antocianinas/análise , Antocianinas/biossíntese , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Flavonoides/análise , Espectroscopia de Ressonância Magnética , Espectrometria de Massas
19.
Food Chem ; 145: 717-24, 2014 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-24128536

RESUMO

Sweetpotato phytochemical content was evaluated in four genotypes (NCPUR06-020, Covington, Yellow Covington, and NC07-847) at harvest and after curing/storage for 4 or 8 months. Curing and storage for up to 8 months did not significantly affect total phenolic content in Covington, Yellow Covington, and NC07-847, however for NCPUR06-020, a purple-fleshed selection, total phenolic content declined mainly due to anthocyanin degradation during storage. Covington had the highest carotenoid content at harvest time (281.9 µg/g DM), followed by NC07-847 (26.2 µg/g DM), and after 8 months, total carotenoids had increased by 25% and 50%, respectively. Antioxidant activity gradually declined during storage, and freshly harvested sweetpotatoes also demonstrated higher anti-inflammatory capacity as gauged by inhibition of lipopolysaccharide-induced reactive oxygen species (ROS) in SH-SY5Y cells. Gradual changes in sweetpotato phytochemical content and antioxidant and anti-inflammatory capacity were noted during normal long-term storage, but the specific effects were genotype-dependent.


Assuntos
Antocianinas/análise , Ácido Ascórbico/análise , Carotenoides/análise , Ipomoea batatas/química , Fenóis/análise , Antocianinas/isolamento & purificação , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Ácido Ascórbico/isolamento & purificação , Carotenoides/isolamento & purificação , Linhagem Celular Tumoral , Sobrevivência Celular , Cromatografia Líquida de Alta Pressão , Armazenamento de Alimentos , Genótipo , Humanos , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Lipopolissacarídeos , Fenóis/isolamento & purificação , Extratos Vegetais/química , Espécies Reativas de Oxigênio/metabolismo , Temperatura , Fatores de Tempo
20.
Brain Res ; 1555: 60-77, 2014 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-24502982

RESUMO

Neuropathological evidence indicates that dopaminergic cell death in Parkinson׳s disease (PD) involves impairment of mitochondrial complex I, oxidative stress, microglial activation, and the formation of Lewy bodies. Epidemiological findings suggest that the consumption of berries rich in anthocyanins and proanthocyanidins may reduce PD risk. In this study, we investigated whether extracts rich in anthocyanins, proanthocyanidins, or other polyphenols suppress the neurotoxic effects of rotenone in a primary cell culture model of PD. Dopaminergic cell death elicited by rotenone was suppressed by extracts prepared from blueberries, grape seed, hibiscus, blackcurrant, and Chinese mulberry. Extracts rich in anthocyanins and proanthocyanidins exhibited greater neuroprotective activity than extracts rich in other polyphenols, and a number of individual anthocyanins interfered with rotenone neurotoxicity. The blueberry and grape seed extracts rescued rotenone-induced defects in mitochondrial respiration in a dopaminergic cell line, and a purple basal extract attenuated nitrite release from microglial cells stimulated by lipopolysaccharide. These findings suggest that anthocyanin- and proanthocyanidin-rich botanical extracts may alleviate neurodegeneration in PD via enhancement of mitochondrial function.


Assuntos
Antocianinas/uso terapêutico , Neurônios Dopaminérgicos/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Doença de Parkinson/tratamento farmacológico , Fitoterapia , Proantocianidinas/uso terapêutico , Rotenona/toxicidade , Animais , Células Cultivadas , Neurônios Dopaminérgicos/metabolismo , Camundongos , Microglia/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Doença de Parkinson/metabolismo , Extratos Vegetais/uso terapêutico , Tirosina 3-Mono-Oxigenase/metabolismo
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