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1.
Int J Food Sci Nutr ; 67(1): 16-9, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26796072

RESUMO

Premature infants are the main recipients of pasteurised donor human milk (PDHM), when their mothers are unable to provide their own. In this study, we evaluated the effect of pasteurisation on the concentrations of vitamin D compounds in donor breastmilk. Milk samples were obtained pre- and post-Holder pasteurisation. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to analyse the samples for vitamins D2 and D3 (D2 and D3) and 25-hydroxyvitamins D2 and D3 (25(OH)D2 and 25(OH)D3). The significance of differences in vitamin D concentrations between the two groups of milk samples was assessed using the Wilcoxon matched-pairs signed rank test, in which p < 0.05 was considered significant. Pasteurisation resulted in a significant reduction (p < 0.05) in the content of D2, D3, 25(OH)D2 and 25(OH)D3. The losses ranged from 10% to 20% following pasteurisation.


Assuntos
Leite Humano/química , Pasteurização , Vitamina D/análise , 25-Hidroxivitamina D 2/análise , Calcifediol/análise , Humanos , Espectrometria de Massas em Tandem , Vitaminas/análise
2.
BMC Genomics ; 16: 561, 2015 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-26220670

RESUMO

BACKGROUND: Mango fruits contain a broad spectrum of phenolic compounds which impart potential health benefits; their biosynthesis is catalysed by enzymes in the phenylpropanoid-flavonoid (PF) pathway. The aim of this study was to reveal the variability in genes involved in the PF pathway in three different mango varieties Mangifera indica L., a member of the family Anacardiaceae: Kensington Pride (KP), Irwin (IW) and Nam Doc Mai (NDM) and to determine associations with gene expression and mango flavonoid profiles. RESULTS: A close evolutionary relationship between mango genes and those from the woody species poplar of the Salicaceae family (Populus trichocarpa) and grape of the Vitaceae family (Vitis vinifera), was revealed through phylogenetic analysis of PF pathway genes. We discovered 145 SNPs in total within coding sequences with an average frequency of one SNP every 316 bp. Variety IW had the highest SNP frequency (one SNP every 258 bp) while KP and NDM had similar frequencies (one SNP every 369 bp and 360 bp, respectively). The position in the PF pathway appeared to influence the extent of genetic diversity of the encoded enzymes. The entry point enzymes phenylalanine lyase (PAL), cinnamate 4-mono-oxygenase (C4H) and chalcone synthase (CHS) had low levels of SNP diversity in their coding sequences, whereas anthocyanidin reductase (ANR) showed the highest SNP frequency followed by flavonoid 3'-hydroxylase (F3'H). Quantitative PCR revealed characteristic patterns of gene expression that differed between mango peel and flesh, and between varieties. CONCLUSIONS: The combination of mango expressed sequence tags and availability of well-established reference PF biosynthetic genes from other plant species allowed the identification of coding sequences of genes that may lead to the formation of important flavonoid compounds in mango fruits and facilitated characterisation of single nucleotide polymorphisms between varieties. We discovered an association between the extent of sequence variation and position in the pathway for up-stream genes. The high expression of PAL, C4H and CHS genes in mango peel compared to flesh is associated with high amounts of total phenolic contents in peels, which suggest that these genes have an influence on total flavonoid levels in mango fruit peel and flesh. In addition, the particularly high expression levels of ANR in KP and NDM peels compared to IW peel and the significant accumulation of its product epicatechin gallate (ECG) in those extracts reflects the rate-limiting role of ANR on ECG biosynthesis in mango.


Assuntos
Flavonoides/biossíntese , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética , Mangifera/genética , Aciltransferases/classificação , Aciltransferases/genética , Anacardiaceae/genética , Anacardiaceae/metabolismo , Sistema Enzimático do Citocromo P-450/classificação , Sistema Enzimático do Citocromo P-450/genética , Etiquetas de Sequências Expressas , Frutas/genética , Frutas/metabolismo , Mangifera/metabolismo , Fenilalanina Amônia-Liase/classificação , Fenilalanina Amônia-Liase/genética , Filogenia , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real , Transcinamato 4-Mono-Oxigenase/classificação , Transcinamato 4-Mono-Oxigenase/genética , Transcriptoma
3.
Bioorg Med Chem Lett ; 24(18): 4523-4528, 2014 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-25172417

RESUMO

Osmolytes have been proposed as treatments for neurodegenerative proteinopathies including Alzheimer's disease. However, for osmolytes to reach the clinic their efficacy must be improved. In this work, copper(I)-catalyzed azide-alkyne cycloaddition chemistry was used to synthesize glycoclusters bearing six copies of trehalose, lactose, galactose or glucose, with the aim of improving the potency of these osmolytes via multivalency. A trehalose glycocluster was found to be superior to monomeric trehalose in its ability to retard the formation of amyloid-beta peptide 40 (Aß40) fibrils and protect neurons from Aß40-induced cell death.


Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Química Click , Descoberta de Drogas , Glicoconjugados/farmacologia , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Fragmentos de Peptídeos/antagonistas & inibidores , Trealose/análogos & derivados , Peptídeos beta-Amiloides/metabolismo , Animais , Relação Dose-Resposta a Droga , Glicoconjugados/síntese química , Glicoconjugados/química , Camundongos , Camundongos Endogâmicos C57BL , Estrutura Molecular , Neurônios/metabolismo , Neurônios/patologia , Fármacos Neuroprotetores/síntese química , Fármacos Neuroprotetores/química , Fragmentos de Peptídeos/metabolismo , Relação Estrutura-Atividade , Trealose/química
4.
Biomed Chromatogr ; 28(9): 1163-6, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24474532

RESUMO

Forty-five strains from two different species (Salinispora arenicola and Salinispora pacifica) were isolated from three different marine sponge species in the Great Barrier Reef region of Australia. We found that two of the strains of Salinispora arenicola (MV0335 and MV0029) produced mevinolin, a fungus-derived cholesterol-lowering agent. Compound structure was determined using an integrated approach: (a) high performance liquid chromatography-quadrupole time-of-flight-mass spectrometric analysis with multimode ionization (electrospray ionization and atmospheric pressure chemical ionization) and fast polarity switching; and (b) database searching and matching of monoisotopic masses, retention times and mass spectra of the precursor and product ions of the compounds of interest and the authentic reference standards thereof.


Assuntos
Lovastatina/química , Lovastatina/metabolismo , Micromonosporaceae/metabolismo , Poríferos/microbiologia , Animais , Anticolesterolemiantes/química , Anticolesterolemiantes/isolamento & purificação , Anticolesterolemiantes/metabolismo , Austrália , Lovastatina/isolamento & purificação , Micromonosporaceae/isolamento & purificação , Espectrometria de Massas por Ionização por Electrospray
5.
Appl Microbiol Biotechnol ; 97(7): 3097-108, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23081774

RESUMO

The developmental cycle of the obligate marine antibiotic producer actinobacterium Salinispora arenicola isolated from a Great Barrier Reef marine sponge was investigated in relation to mycelium and spore ultrastructure, synthesis of rifamycin antibiotic compounds, and expression of genes correlated with spore formation and with rifamycin precursor synthesis. The developmental cycle of S. arenicola M413 on solid agar medium was characterized by substrate mycelium growth, change of colony color, and spore formation; spore formation occurred quite early in colony growth but development of black colonies occurred only at late stages, correlated with a change in spore maturity in relation to cell wall layers. Rifamycins were detected throughout the growth cycle, but changed in relative quantity at particular phases in the cycle, with a marked increase after 32 days. Expression of the spore division gene ssgA and the rifK gene for 3-amino-5-hydroxybenzoate synthase responsible for rifamycin precursor synthesis was seen even at early stages of the growth cycle. ssgA expression significantly increased between days 26 and 31, but rifK expression effectively remained constant throughout the growth cycle, consistent with the early synthesis of rifamycin. Factors other than precursor synthesis may be responsible for an observed late increase in rifamycin production. A useful approach for measuring and exploring the regulation of antibiotic synthesis and gene expression in the marine natural product producer S. arenicola has been established.


Assuntos
Antibacterianos/metabolismo , Micromonosporaceae/crescimento & desenvolvimento , Micromonosporaceae/metabolismo , Poríferos/microbiologia , Rifamicinas/metabolismo , Animais , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/genética , Vias Biossintéticas/genética , Proteínas de Ciclo Celular/biossíntese , Proteínas de Ciclo Celular/genética , Meios de Cultura/química , Perfilação da Expressão Gênica , Micromonosporaceae/isolamento & purificação , Micromonosporaceae/ultraestrutura , Pigmentos Biológicos/metabolismo , Esporos Bacterianos/ultraestrutura , Fatores de Tempo
6.
AAPS PharmSciTech ; 14(1): 301-11, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23307066

RESUMO

The controlled release of diflunisal and fluconazole from tablets made of novel polymers, poly(acrylic acid) (PAA) crosslinked with either ß-cyclodextrin (ßCD) or hydroxypropyl-ßCD (HPßCD), was investigated and Carbopol 934P (Carbopol) was used as a highly crosslinked PAA for comparison. Diflunisal strongly associates with ßCD-PAA and HPßCD-PAA polymers (Ka of 486 and 6,055 M(-1) respectively); thus, it was physically mixed into the conjugates and also precomplexed to identify whether decomplexation has any influence on release kinetics. Fluconazole has poor complexing ability (Ka of 34 M(-1) with HPßCD-PAA); thus, it was only tested as a physical mixture. Swelling and adhesion studies were conducted on all tablet combinations and adhesivity of the CD-PAA polymer tablets was maintained. Diflunisal release was much slower from HPßCD-PAA tablets than from ßCD-PAA, suggesting that a higher degree of complexation retards release. The precomplexed diflunisal release was also slower than the physically mixed diflunisal of the corresponding conjugate. The release closely followed zero-order kinetics for HPßCD-PAA, but was more sigmoidal for ßCD-PAA and especially Carbopol. Conversely, poorly associating fluconazole released in almost exactly the same way across both polymers and Carbopol, indicating that the release kinetics of poorly associating drugs are not influenced by the presence of cyclodextrins. In view of the varying profiles and release rates shown with diflunisal for the different polymers, the fluconazole data support the concept that adequate complexation can indeed modulate the release kinetics of drugs.


Assuntos
Resinas Acrílicas/química , Ciclodextrinas/química , Diflunisal/administração & dosagem , Formas de Dosagem , Fluconazol/administração & dosagem , Preparações de Ação Retardada
7.
Int J Hyg Environ Health ; 253: 114237, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37544043

RESUMO

Environmental exposure to multiple metals and metalloids is widespread, leading to a global concern relating to the adverse health effects of mixed-metals exposure, especially in young children living around industrial areas. This study aimed to quantify the concentrations of essential and potentially toxic elements in blood and to examine the potential associations between multiple elements exposures, growth determinants, and liver and kidney function biomarkers in children living in several industrial areas in Dhaka, Bangladesh. The blood distribution of 20 trace elements As, Ag, Bi, Br, Cd, Co, Cr, Cu, I, Mn, Hg, Mo, Ni, Pb, Se, Sb, Tl, V, U, and Zn, growth determinants such as body mass index and body fats, blood pressure, liver and kidney injury biomarkers including serum alanine aminotransferase and alkaline phosphatase activities, serum calcium, and creatinine levels, blood urea nitrogen, and hemoglobin concentrations, and glomerular filtration rate were measured in 141 children, aged six to 16 years. Among these elements, blood concentrations of Ag, U, V, Cr, Cd, Sb, and Bi were measured below LOQs and excluded from subsequent statistical analysis. This comprehensive study revealed that blood concentrations of these elements in children, living in industrial areas, exceeded critical reference values to varying extents; elevated exposure to As, Pb, Br, Cu, and Se was found in children living in multiple industrial areas. A significant positive association between elevated blood Tl concentration and obesity (ß = 0.300, p = 0.007) and an inverse relationship between lower As concentration and underweight (ß = -0.351, p < 0.001) compared to healthy weight children indicate that chronic exposure to Tl and As may influence the metabolic burden and physical growth in children. Concentration-dependent positive associations were observed between the blood concentrations of Cu, Se, and Br and hepatic- and renal dysfunction biomarkers, an inverse association with blood Mo and I level, however, indicates an increased risk of Cu, Se, and Br-induced liver and kidney toxicity. Further in-depth studies are warranted to elucidate the underlying mechanisms of the observed associations. Regular biomonitoring of elemental exposures is also indispensable to regulate pollution in consideration of the long-term health effects of mixed-elements exposure in children.


Assuntos
Cádmio , Oligoelementos , Humanos , Criança , Pré-Escolar , Cádmio/análise , Chumbo/análise , Bangladesh , Oligoelementos/metabolismo , Fígado/química , Fígado/metabolismo , Rim/química
8.
J Anal Toxicol ; 44(9): 1036-1046, 2021 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-32232355

RESUMO

Essential and nonessential element concentrations in human blood provide important information on the nutritional status of individuals and can assist in the screening or diagnosis of certain disorders and their association with other causative factors. A simple and sensitive method, suitable for use with small sample volumes, for quantification of multiple trace element concentrations in whole blood and plasma has been developed using inductively coupled plasma-mass spectrometry. Method validation was performed using standard reference materials of whole blood and serum using varying sample treatments with nitric acid, water and hydrogen peroxide. The method was applied to quantify the trace element concentrations in whole blood and plasma samples (0.1 mL) from 50 adult blood donors in Queensland. The whole blood sample (5 mL) was collected in Vacutainer tubes with K2EDTA as anticoagulant. The developed method was able to quantify, in blood and plasma samples over a wide range of concentrations, several essential elements: cobalt, copper, zinc, iron, manganese and selenium; the nutritionally probably essential elements vanadium and strontium; and nonessential elements including lead, cadmium, arsenic, caesium, barium, thallium and uranium. Significant differences (P < 0.0001) were observed between whole blood and plasma concentrations for 13 elements; 5 of the measured elements, cobalt (0.49 vs. 0.36 µg/L), copper (1.0 vs. 0.75 mg/L), strontium (28 vs. 16 µg/L), barium (1.5 vs. 0.64 µg/L) and thallium (0.06 vs. 0.03 µg/L), had higher mean concentrations in plasma than in blood. Whole blood concentrations of nine trace elements were significantly correlated (P < 0.0001) with plasma concentrations. The distribution of the trace elements between human blood and plasma varied considerably for the different elements. These results indicate that, using a small sample volume, this assay is suitable for the evaluation of nutritional status as well as in monitoring human toxic elemental exposures.


Assuntos
Espectrometria de Massas , Oligoelementos/sangue , Adulto , Cádmio , Cobre , Humanos , Plasma , Selênio , Análise Espectral , Zinco
9.
J Trace Elem Med Biol ; 68: 126804, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34111708

RESUMO

BACKGROUND: Knowledge of trace element stability during sample handling and preservation is a prerequisite to produce reliable test results in clinical trace element analysis. METHOD: An alkaline dissolution method has been developed using inductively coupled plasma mass spectrometry to quantify eighteen trace element concentrations: vanadium, chromium, manganese, cobalt, nickel, copper, zinc, arsenic, selenium, bromine, molybdenum, cadmium, antimony, iodine, mercury, thallium, lead, and bismuth in human blood, using a small sample volume of 0.1 mL. The study evaluated the comparative effects of storage conditions on the stability of nutritionally essential and non-essential elements in human blood and plasma samples stored at three different temperatures (4 °C, -20 °C and -80 °C) over a one-year period, and analysed at multiple time points. The distribution of these elements between whole blood and plasma and their distribution relationships are illustrated using blood samples from 66 adult donors in Queensland. RESULTS: The refrigeration and freezing of blood and plasma specimens proved to be suitable storage conditions for many of the trace elements for periods up to six months, with essentially unchanged concentrations. Substantially consistent recoveries were obtained by preserving specimens at -20 °C for up to one year. Ultra-freezing of the specimens at -80 °C did not improve stability; but appeared to result in adsorption and/or precipitation of some elements, accompanied by a longer sample thawing time. A population sample study revealed significant differences between the blood and plasma concentrations of six essential elements and their relationships also varied significantly for different elements. CONCLUSION: Blood and plasma specimens can be reliably stored at 4 °C for six months or kept frozen at -20 °C up to one year to obtain high quality test results of trace elements.


Assuntos
Selênio , Oligoelementos , Adulto , Cádmio , Cromo , Humanos , Zinco
10.
J Agric Food Chem ; 56(9): 3037-42, 2008 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-18393431

RESUMO

Mangos are a source of bioactive compounds with potential health-promoting activity. This study evaluated the abilities of the mango components quercetin and mangiferin and the aglycone derivative of mangiferin, norathyriol, to modulate the transactivation of peroxisome proliferator-activated receptor isoforms (PPARs). PPARs are transcription factors important in many human diseases. Through the use of a gene reporter assay it was shown that quercetin inhibited the activation of all three isoforms of PPARs (PPARgamma IC(50) = 56.3 microM; PPARalpha IC(50) = 59.6 microM; PPARbeta IC(50) = 76.9 microM) as did norathyriol (PPARgamma IC(50) = 153.5 microM; PPARalpha IC(50) = 92.8 microM; PPARbeta IC(50) = 102.4 microM), whereas mangiferin did not inhibit the transactivation of any isoform. These findings suggest that mango components and metabolites may alter transcription and could contribute to positive health benefits via this or similar mechanisms.


Assuntos
Mangifera/química , Receptores Ativados por Proliferador de Peroxissomo/genética , Quercetina/farmacologia , Ativação Transcricional/efeitos dos fármacos , Xantenos/farmacologia , Xantonas/farmacologia , Frutas/química , Humanos , Isoformas de Proteínas/genética , Transfecção
11.
J Chromatogr B Analyt Technol Biomed Life Sci ; 852(1-2): 362-6, 2007 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-17301003

RESUMO

HPLC-MS-MS has been used for the identification and characterisation of rifamycin B and rifamycin SV in various strains of the marine sponge-derived bacterium Salinispora. Gradient elution using acetonitrile/water/ammonium acetate was used to separate the rifamycins from the matrix and negative ion-electrospray mass spectrometry was used for detection and confirmation. The presence of rifamycin in bacterial extracts was confirmed by matching retention times, parent ion spectra and the fragmentated parent ion spectra of the standard compounds and the bacterial extracts. All strains of the marine sponge bacterium Salinispora tested were found to contain rifamycin thus an alternate actinobacterial source of rifamycin was established.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Bactérias Gram-Positivas/metabolismo , Poríferos/microbiologia , Rifamicinas/biossíntese , Espectrometria de Massas em Tandem/métodos , Animais
12.
J Steroid Biochem Mol Biol ; 171: 262-269, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28428023

RESUMO

Cytochrome P450s (CYP450s) are a rapidly evolving family of enzymes, making it difficult to identify bona fide orthologs with notable lineage-specific exceptions. In ecdysozoans, a small number of the most conserved orthologs include enzymes which metabolize ecdysteroids. Ecdysone pathway components were recently shown in a decapod crustacean but with a notable absence of shade, which is important for converting ecdysone to its active form, 20-hydroxyecdysone (20HE), suggesting that another CYP450 performs a similar function in crustaceans. A CYPome temporal expression analysis throughout metamorphosis performed in this research highlights several un-annotated CYP450s displaying differential expression and provides information into expression patterns of annotated CYP450s. Using the expression patterns in the Eastern spiny lobster Sagmariasus verreauxi, followed by 3D modelling and finally activity assays in vitro, we were able to conclude that a group of CYP450s, conserved across decapod crustaceans, function as the insect shade. To emphasize the fact that these genes share the function with shade but are phylogenetically distinct, we name this enzyme system Shed.


Assuntos
Proteínas de Artrópodes/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Metamorfose Biológica , Modelos Moleculares , Palinuridae/enzimologia , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Células COS , Chlorocebus aethiops , Biologia Computacional , Sistema Enzimático do Citocromo P-450/química , Sistema Enzimático do Citocromo P-450/genética , Bases de Dados de Compostos Químicos , Bases de Dados Genéticas , Bases de Dados de Proteínas , Ecdisterona/química , Ecdisterona/metabolismo , Perfilação da Expressão Gênica , Hidroxilação , Anotação de Sequência Molecular , Estrutura Molecular , Peso Molecular , Palinuridae/crescimento & desenvolvimento , Filogenia , Conformação Proteica , Proteínas Recombinantes/metabolismo
13.
FEMS Microbiol Lett ; 364(2)2017 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-28087611

RESUMO

The structures of acyl homoserine lactone (AHL) compounds and their quantification were accomplished using an integrated liquid chromatography-mass spectrometry approach. The precursor and product ions, along with retention times of peaks, were searched against an in-house database of AHLs and structures confirmed by accurate mass and by comparison with authentic AHL standards. The two compounds, N-(3-oxodecanoyl)-L-homoserine lactone and N-(3-oxododecanoyl)-L-homoserine lactone, were characterised and quantified in Salinispora sp. cultures.


Assuntos
Acil-Butirolactonas/análise , Organismos Aquáticos/metabolismo , Micromonosporaceae/metabolismo , Poríferos/microbiologia , Animais , Organismos Aquáticos/química , Organismos Aquáticos/isolamento & purificação , Cromatografia Líquida , Meios de Cultura/química , Espectrometria de Massas , Micromonosporaceae/química , Micromonosporaceae/isolamento & purificação
14.
Artigo em Inglês | MEDLINE | ID: mdl-16931178

RESUMO

A quantitative liquid chromatography positive ion electrospray tandem mass spectrometric method for the simultaneous determination of sulforaphane, iberin and their metabolites in human urine and plasma is described. The stability of the metabolites was determined in aqueous solution and in human plasma. Gradient liquid chromatographic separation was performed on a Zorbax SB-Aq 3.5 microm (100 x 2.1mm) column, using a mobile phase (flow rate 0.25 mL/min) consisting of ammonium acetate buffer at pH 4 and acetonitrile. Butyl thiocarbamoyl l-cysteine was used as internal standard. The assay was linear (r(2)>0.99) over the range of 0.03-300 microM in urine and 0.03-15 microM in plasma with intra- and inter-day assay precision (<10% CV) and accuracy (<20%). The lower limits of quantitation were in the range of 10-150 nmol/L. The method has been used to report, for the first time, individual quantitative measurement of each of the mercapturic acid pathway metabolites of sulforaphane and iberin in both human plasma and urine following a dietary study of broccoli consumption.


Assuntos
Acetilcisteína/metabolismo , Cromatografia Líquida/métodos , Isotiocianatos/metabolismo , Espectrometria de Massas por Ionização por Electrospray/métodos , Tiocianatos/metabolismo , Acetilcisteína/sangue , Acetilcisteína/urina , Humanos , Isotiocianatos/sangue , Isotiocianatos/urina , Estrutura Molecular , Reprodutibilidade dos Testes , Sulfóxidos , Tiocianatos/sangue , Tiocianatos/urina
15.
Curr Drug Deliv ; 3(4): 389-97, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17076641

RESUMO

ISCOMs have received much attention as vaccine adjuvants due to their immunostimulatory effects. They are colloidal particles typically comprised of phospholipids, cholesterol and Quil A, a crude mixture of saponins extracted from the bark of Quillaja saponaria Molina. We have previously shown that ISCOMs can be prepared by ether injection wherein an ether solution of phospholipids and cholesterol in a mass ratio of 5:2 is injected into a solution of Quil A at a mass ratio of 7 lipids: 3 Quil A. The aim of this study was firstly to isolate and characterise discrete fractions of Quil A and secondly to investigate which of these fractions were able to form ISCOMs by the method of ether injection. Six fractions of Quil A were isolated by semi-preparative reverse phase high performance liquid chromatography (RP-HPLC) and characterised by analytical HPLC, liquid chromatography tandem mass spectrometry (LC-MS) and the qualitative Liebermann-Burchard and Molisch tests for triterpenoids and carbohydrates respectively. ISCOMs were subsequently prepared from the isolated fractions by the method of ether injection and the resulting preparations characterized by photon correlation spectroscopy (PCS) and negative stain transmission electron microscopy (TEM). The molecular weights of the major compounds in the fractions ranged from approximately 1200 to approximately 2300 Da; all fractions tested positive for triterpenoids and saccharides and four of the fractions were identified as QS-7, QS-17, QS-18 and QS-21 by analysis (LC-MS and analytical HPLC). Injection of ether solutions of lipids into aqueous solutions of QS-17, QS-18 or QS-21 all resulted in homogeneous ISCOM dispersions. The combination of lipids and QS-7 by ether injection produced lamellae and liposomes as the prominent structures and a minor amount of ISCOMs. The remaining two hydrophilic, low molecular weight fractions of Quil A did not produce ISCOMs, instead liposomes and helical structures predominated in the samples.


Assuntos
ISCOMs/química , Lipídeos/química , Quillaja/química , Saponinas/química , Lipossomos , Estrutura Molecular , Peso Molecular , Tamanho da Partícula , Saponinas de Quilaia , Saponinas/isolamento & purificação
16.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1009-1010: 80-6, 2016 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-26708628

RESUMO

The determination of both the water-soluble and lipid-soluble vitamin D compounds in human biological fluids is necessary to illuminate potentially significant biochemical mechanisms. The lack of analytical methods to quantify the water-soluble forms precludes studies on their role and biological functions; currently available liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods are able to determine only a single sulfated form of Vitamin D. We describe here a highly sensitive and specific LC-MS/MS method for the quantification of four sulfated forms of vitamin D: vitamins D2- and D3-sulfate (D2-S and D3-S) and 25-hydroxyvitamin D2- and D3-sulfate (25(OH)D2-S and 25(OH)D3-S). A comparative evaluation showed that the ionization efficiencies of underivatized forms in negative ion mode electrospray ionisation (ESI) are superior to those of the derivatized (using 4-phenyl-l,2,4-triazoline-3,5-dione (PTAD)) forms in positive ion mode ESI. Separation was optimised to minimise co-elution with endogenous matrix compounds, thereby reducing ion suppression/enhancement effects. Isotopically labelled analogues of each compound were used as internal standards to correct for ion suppression/enhancement effects. The method was validated and then applied for the analysis of breastmilk and human serum. The detection limits, repeatability standard deviations, and recoveries ranged from 0.20 to 0.28fmol, 2.8 to 10.2%, and 81.1 to 102%, respectively.


Assuntos
Colecalciferol/análogos & derivados , Cromatografia Líquida/métodos , Ergocalciferóis/análise , Ergocalciferóis/sangue , Leite Humano/química , Espectrometria de Massas em Tandem/métodos , 25-Hidroxivitamina D 2/análise , 25-Hidroxivitamina D 2/sangue , Animais , Colecalciferol/análise , Colecalciferol/sangue , Feminino , Humanos , Limite de Detecção , Espectrometria de Massas por Ionização por Electrospray/métodos
17.
J Immunotoxicol ; 13(4): 590-602, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27416522

RESUMO

Chronic inflammation is linked with the generation and progression of various diseases such as cancer, diabetes and atherosclerosis, and anti-inflammatory drugs therefore have the potential to assist in the treatment of these conditions. Carica papaya is a tropical plant that is traditionally used in the treatment of various ailments including inflammatory conditions. A literature search was conducted by using the keywords "papaya", "anti-inflammatory and inflammation" and "immunomodulation and immune" along with cross-referencing. Both in vitro and in vivo investigation studies were included. This is a review of all studies published since 2000 on the anti-inflammatory activity of papaya extracts and their effects on various immune-inflammatory mediators. Studies on the anti-inflammatory activities of recognized phytochemicals present in papaya are also included. Although in vitro and in vivo studies have shown that papaya extracts and papaya-associated phytochemicals possess anti-inflammatory and immunomodulatory properties, clinical studies are lacking.


Assuntos
Anti-Inflamatórios/uso terapêutico , Carica/imunologia , Fatores Imunológicos/uso terapêutico , Inflamação/terapia , Fitoterapia , Extratos Vegetais/uso terapêutico , Animais , Humanos
18.
Food Chem ; 199: 185-94, 2016 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-26775960

RESUMO

Various food constituents have been proposed as disease-modifying agents for Alzheimer's disease (AD), due to epidemiological evidence of their beneficial effects, and for their ability to ameliorate factors linked to AD pathogenesis, namely by: chelating iron, copper and zinc; scavenging reactive oxygen species; and suppressing the fibrillation of amyloid-beta peptide (Aß). In this study, nine different food constituents (l-ascorbic acid, caffeic acid, caffeine, curcumin, (-)-epigallocatechin gallate (EGCG), gallic acid, propyl gallate, resveratrol, and α-tocopherol) were investigated for their effects on the above factors, using metal chelation assays, antioxidant assays, and assays of Aß42 fibrillation. An assay method was developed using 5-Br-PAPS to examine the complexation of Zn(II) and Cu(II). EGCG, gallic acid, and curcumin were identified as a multifunctional compounds, however their poor brain uptake might limit their therapeutic effects. The antioxidants l-ascorbic acid and α-tocopherol, with better brain uptake, deserve further investigation for specifically addressing oxidative stress within the AD brain.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Peptídeos beta-Amiloides/antagonistas & inibidores , Quelantes/farmacologia , Sequestradores de Radicais Livres/farmacologia , Metais/metabolismo , Fragmentos de Peptídeos/antagonistas & inibidores , Encéfalo/metabolismo , Catequina/análogos & derivados , Catequina/farmacologia , Curcumina/farmacologia , Ácido Gálico/farmacologia , Humanos
19.
Anal Chim Acta ; 891: 211-20, 2015 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-26388380

RESUMO

Milk is an important source of nutrients for various risk populations, including infants. The accurate measurement of vitamin D in milk is necessary to provide adequate supplementation advice for risk groups and to monitor regulatory compliance. Currently used liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods are capable of measuring only four analogues of vitamin D in unfortified milk. We report here an accurate quantitative analytical method for eight analogues of vitamin D: Vitamin D2 and D3 (D2 and D3), 25-hydroxy D2 and D3, 24,25-dihydroxy D2 and D3, and 1,25-dihydroxyD2 and D3. In this study, we compared saponification and protein precipitation for the extraction of vitamin D from milk and found the latter to be more effective. We also optimised the pre-column derivatisation using 4-phenyl-l,2,4-triazoline-3,5-dione (PTAD), to achieve the highest sensitivity and accuracy for all major vitamin D forms in milk. Chromatography was optimised to reduce matrix effects such as ion-suppression, and the matrix effects were eliminated using co-eluting stable isotope labelled internal standards for the calibration of each analogue. The analogues, 25-hydroxyD3 (25(OH)D3) and its epimer (3-epi-25(OH)D3) were chromatographically resolved, to prevent over-estimation of 25(OH)D3. The method was validated and subsequently applied for the measurement of total vitamin D levels in human, cow, mare, goat and sheep milk samples. The detection limits, repeatability standard deviations, and recovery ranges were from 0.2 to 0.4 femtomols, 6.30-13.5%, and 88.2-105%, respectively.


Assuntos
Leite/química , Espectrometria de Massas em Tandem/métodos , Vitamina D/análogos & derivados , Vitamina D/análise , Animais , Bovinos , Cromatografia Líquida/métodos , Cabras , Humanos , Limite de Detecção , Ovinos
20.
Food Funct ; 6(6): 1847-54, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25940566

RESUMO

Mango fruit contain many bioactive compounds, some of which are transcription factor regulators. Estrogen receptor alpha (ERα) and beta (ERß) are two regulators of gene transcription that are important in a variety of physiological processes and also in diseases including breast cancer. We examined the ability of the mango constituents quercetin, mangiferin, and the aglycone form of mangiferin, norathyriol, to activate both isoforms of the estrogen receptor. Quercetin and norathyriol decreased the viability of MCF-7 breast cancer cells whereas mangiferin had no effect on MCF-7 cells. We also determined that quercetin and mangiferin selectively activated ERα whereas norathyriol activated both ERα and ERß. Despite quercetin, mangiferin and norathyriol having similar polyphenolic structural motifs, only norathyriol activated ERß, showing that bioactive agents in mangoes have very specific biological effects. Such specificity may be important given the often-opposing roles of ERα and ERß in breast cancer proliferation and other cellular processes.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Receptor alfa de Estrogênio/agonistas , Fitoestrógenos/farmacologia , Quercetina/farmacologia , Xantenos/farmacologia , Xantonas/farmacologia , Animais , Antineoplásicos Fitogênicos/antagonistas & inibidores , Antineoplásicos Fitogênicos/metabolismo , Neoplasias da Mama/metabolismo , Células COS , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , Antagonistas do Receptor de Estrogênio/farmacologia , Receptor alfa de Estrogênio/antagonistas & inibidores , Receptor alfa de Estrogênio/genética , Receptor alfa de Estrogênio/metabolismo , Receptor beta de Estrogênio/agonistas , Receptor beta de Estrogênio/antagonistas & inibidores , Receptor beta de Estrogênio/genética , Receptor beta de Estrogênio/metabolismo , Feminino , Frutas/química , Genes Reporter/efeitos dos fármacos , Humanos , Células MCF-7 , Mangifera/química , Proteínas de Neoplasias/agonistas , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Fitoestrógenos/antagonistas & inibidores , Fitoestrógenos/metabolismo , Quercetina/antagonistas & inibidores , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Elementos de Resposta/efeitos dos fármacos , Ativação Transcricional/efeitos dos fármacos , Xantenos/antagonistas & inibidores , Xantenos/metabolismo , Xantonas/antagonistas & inibidores , Xantonas/metabolismo
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