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1.
J Allergy Clin Immunol ; 153(3): 742-758, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38042501

RESUMO

BACKGROUND: Food allergy (FA) is one of the most common chronic conditions in children with an increasing prevalence facilitated by the exposure to environmental factors in predisposed individuals. It has been hypothesized that the increased consumption of ultra-processed foods, containing high levels of dietary advanced glycation end products (AGEs), could facilitate the occurrence of FA. OBJECTIVE: We sought to provide preclinical and clinical evidence on the potential role of AGEs in facilitating the occurrence of FA. METHODS: Human enterocytes, human small intestine organ culture, and PBMCs from children at risk for allergy were used to investigate the direct effect of AGEs on gut barrier, inflammation, TH2 cytokine response, and mitochondrial function. Intake of the 3 most common glycation products in Western diet foods, Nε-(carboxymethyl) lysine, Nε-(1-carboxyethyl) lysin, and Nδ-(5-hydro-5- methyl-4-imidazolone-2-yl)-ornithine (MG-H1), and the accumulation of AGEs in the skin were comparatively investigated in children with FA and in age-matched healthy controls. RESULTS: Human enterocytes exposed to AGEs showed alteration in gut barrier, AGE receptor expression, reactive oxygen species production, and autophagy, with increased transepithelial passage of food antigens. Small intestine organ cultures exposed to AGEs showed an increase of CD25+ cells and proliferating crypt enterocytes. PBMCs exposed to AGEs showed alteration in proliferation rate, AGE receptor activation, release of inflammatory and TH2 cytokines, and mitochondrial metabolism. Significant higher dietary AGE intake and skin accumulation were observed children with FA (n = 42) compared with age-matched healthy controls (n = 66). CONCLUSIONS: These data, supporting a potential role for dietary AGEs in facilitating the occurrence of FA, suggest the importance of limiting exposure to AGEs children as a potential preventive strategy against this common condition.


Assuntos
Produtos Finais da Glicação Avançada em Alimentos , Hipersensibilidade Alimentar , Criança , Humanos , Receptor para Produtos Finais de Glicação Avançada , Produtos Finais de Glicação Avançada/metabolismo , Dieta Ocidental , Dieta
2.
Molecules ; 28(7)2023 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-37049871

RESUMO

The lack of a practical "fit for the purpose" analytical protocol is the main limitation that has hampered the exploitation of the EFSA analytical health claim on the extra virgin olive oil (EVOO) biophenols, more than ten years since its introduction. In this work, two analytical methods recently developed in our laboratories for categorizing EVOO have been evaluated on a set of 16 samples from Cilento (Campania Region, southern Italy) and compared to other commonly used quality indexes. The Coulometrically Determined Antioxidant Capacity (CDAC) is associated with the component responsible for the health-promoting properties and oxidative stability of EVOO. The Fast Blue BB (FBBB) assay consists of the spectrophotometric (420 nm) determination of biophenols-FBBB diazonium coupling products generated in unfractionated EVOO. The FBBB assay and HPLC-UV reference method provide values highly correlated to each other. Fourteen of sixteen EVOO samples with CDAC > 10 mmol kg-1 and FBBB absorbance > 0.5 had HPLC-determined biophenols > 250 mg kg-1, and therefore eligible for the EFSA health claim. Consistently, two EVOO samples with HPLC-determined biophenols < 250 mg kg-1 had CDAC values and FBBB absorbance below the respective thresholds. CDAC and FBBB assays are proposed individually or in combination as methods to categorize EVOO samples in alternative to HPLC-UV.


Assuntos
Antioxidantes , Azeite de Oliva , Fenóis , Espectrofotometria , Azeite de Oliva/química , Azeite de Oliva/metabolismo , Fenóis/análise , Fenóis/química , Análise de Componente Principal , Ácidos Graxos/análise , Ácidos Graxos/química , Antioxidantes/análise , Antioxidantes/metabolismo , Fatores de Tempo
3.
J Proteome Res ; 21(5): 1330-1339, 2022 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-35347988

RESUMO

Multiple analytical techniques were combined to achieve a detailed characterization of organic residues in different typologies of funerary pottery, which were found at two separate archeological sites in the Campania Region (Italy) and both dated back to the first millennium BC. Gas chromatography-mass spectrometry (GC-MS) analysis of lipids provided inconclusive results. The attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectra of encrustation on two glazed bowls of the 3rd to 4th century BC were comparable to those of fresh bone, revealing the presence of hydroxyapatite and proteins, which were identified as bovine collagen chains by liquid chromatography coupled to high-resolution tandem mass spectrometry (LC-MS/MS)-based proteomics. This finding confirmed that Italic populations used to inhume the dead along with votive meat offerings. Proteomics was decisive for identifying bovine milk in an unusually shaped amphora unearthed from a grave that belonged to a woman at the necropolis of the Greek colony in Cuma (7th century BC). Peptidomic analysis demonstrated that the genetic variant A1 of ß-casein was already present in the southern Mediterranean area at least 2500 years ago. Overall, these results depict an agropastoral system of Italic populations at the age of Magna Graecia based on a significant role of domesticated cattle.


Assuntos
Proteômica , Espectrometria de Massas em Tandem , Animais , Caseínas/análise , Bovinos , Cromatografia Líquida , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos
4.
Crit Rev Food Sci Nutr ; 62(31): 8686-8702, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34060381

RESUMO

Current approaches based on electrophoretic, chromatographic or immunochemical principles have allowed characterizing multiple allergens, mapping their epitopes, studying their mechanisms of action, developing detection and diagnostic methods and therapeutic strategies for the food and pharmaceutical industry. However, some of the common structural features related to the allergenic potential of food proteins remain unknown, or the pathological mechanism of food allergy is not yet fully understood. In addition, it is also necessary to evaluate new allergens from novel protein sources that may pose a new risk for consumers. Technological development has allowed the expansion of advanced technologies for which their whole potential has not been entirely exploited and could provide novel contributions to still unexplored molecular traits underlying both the structure of food allergens and the mechanisms through which they sensitize or elicit adverse responses in human subjects, as well as improving analytical techniques for their detection. This review presents cutting-edge instrumental techniques recently applied when studying structural and functional aspects of proteins, mechanism of action and interaction between biomolecules. We also exemplify their role in the food allergy research and discuss their new possible applications in several areas of the food allergy field.


Assuntos
Alérgenos , Hipersensibilidade Alimentar , Humanos , Alérgenos/química , Hipersensibilidade Alimentar/terapia , Epitopos
5.
Int J Mol Sci ; 23(3)2022 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-35162980

RESUMO

Protein expression from the berry skin of four red grape biotypes with varying hybrid character was compared at a proteome-wide level to identify the metabolic pathways underlying divergent patterns of secondary metabolites. A bottom-up shotgun proteomics approach with label-free quantification and MaxQuant-assisted computational analysis was applied. Red grapes were from (i) purebred Vitis vinifera (Aglianico cv.); (ii) V. vinifera (local Sciascinoso cv.) grafted onto an American rootstock; (iii) interspecific hybrid (V. vinifera × V. labrusca, Isabel), and (iv) uncharacterized grape genotype with hybrid lineage, producing relatively abundant anthocyanidin 3,5-O-diglucosides. Proteomics supported the differences between hybrids and purebred V. vinifera grapes, consistently with distinct phenotypic metabolite assets. Methanol O-anthraniloyltransferase, which catalyses the synthesis of methyl anthranilate, primarily responsible for the "foxy" odour, was exclusive of the Isabel hybrid grape. Most of the proteins with different expression profiles converged into coordinated biosynthetic networks of primary metabolism, while many possible enzymes of secondary metabolism pathways, including 5-glucosyltransferases expected for hybrid grapes, remained unassigned due to incomplete protein annotation for the Vitis genus. Minor differences of protein expression distinguished V. vinifera scion grafted onto American rootstocks from purebred V. vinifera skin grapes, supporting a slight influence of the rootstock on the grape metabolism.


Assuntos
Vitis , Antocianinas/metabolismo , Frutas/genética , Frutas/metabolismo , Odorantes/análise , Proteômica , Vitis/metabolismo
6.
Molecules ; 27(4)2022 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-35209097

RESUMO

Tritordeum results from the crossbreeding of a wild barley (Hordeum chilense) species with durum wheat (Triticum turgidum spp. turgidum). This hexaploid crop exhibits agronomic and rheological characteristics like soft wheat, resulting in an innovative raw material to produce baked goods. We applied a gel-based proteomic approach on refined flours to evaluate protein expression differences among two widespread tritordeum cultivars (Aucan and Bulel) taking as the reference semolina and flour derived from a durum and a soft wheat cvs, respectively. The products of in vitro digestion of model breads were analyzed to compare bio-accessibility of nutrients and mapping tritordeum bread resistant peptides. Significant differences among the protein profiles of the four flours were highlighted by electrophoresis. The amino acid bio-accessibility and the reducing sugars of tritordeum and wheat breads were comparable. Tritordeum cvs had about 15% higher alpha-amino nitrogen released at the end of the duodenal simulated digestion than soft wheat (p < 0.05). Bulel tritordeum flour, bread and digested bread had about 55% less R5-epitopes compared to the soft wheat. Differences in protein expression found between the two tritordeum cvs reflected in diverse digestion products and allergenic and celiacogenic potential of the duodenal peptides. Proteomic studies of a larger number of tritordeum cvs may be successful in selecting those with good agronomical performances and nutritional advantages.


Assuntos
Pão/análise , Grão Comestível/química , Análise de Alimentos , Triticum/química , Cromatografia Líquida , Digestão , Peptídeos/análise , Proteínas de Vegetais Comestíveis/análise , Proteômica/métodos , Espectrometria de Massas em Tandem
7.
J Food Sci Technol ; 58(3): 921-930, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33678875

RESUMO

Pistachio flour obtained from oil industry was nutritionally characterized for use as food ingredient in functional foods. Proximal composition, jointly with mineral content, amino acids and fatty acid profile were studied. In addition, different components present in this food ingredient have been analyzed by attenuated total reflectance Fourier transform infrared spectroscopy and thermal properties of proteins were evaluated by differential scanning calorimetry. This flour presented high mineral content such as potassium, phosphorus, magnesium and calcium. Moreover, high amount of unsaturated fatty acids, mainly oleic and linoleic were found. Secondary structure of proteins mainly was formed by parallel ß-sheet and α-helix. In the by-product, pistachio protein is in a native state and is able to be denatured at temperatures higher than 100 °C. Therefore, food processing of this ingredient can affect the structure of components.

8.
Int J Food Sci Nutr ; 71(4): 410-423, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31544542

RESUMO

Total phenolic content and antioxidant activity of polar extracts of edible resources from Fedora hemp cultivar (Cannabis sativa L.), namely seed, flour and oil, were evaluated. The main components in the polar extracts were identified using HPLC-DAD and HPLC-ESI-MS/MS. As expected, the molecular profile of components from seeds and flour was strictly similar, dominated by N-trans-caffeoyltyramine. The profile of oil polar extracts contained hydroxycinnamic acid derivatives and cannabinoids at lower extent. While the extracts from hemp seed and flour did not interfere with growth of Caco-2 and HT-29 cell, the one from oil (150 µg/mL) significantly reduced cell viability after 24 h of treatment. This effect was associated with the activation of apoptotic cell death and was independent from the antioxidant capacity of the oil polar extract. Notably, HT-29 cells differentiated with sodium butyrate were not sensitive to the cytotoxic effect of the oil extract.


Assuntos
Antioxidantes/farmacologia , Cannabis/química , Neoplasias Colorretais/tratamento farmacológico , Extratos Vegetais/farmacologia , Apoptose/efeitos dos fármacos , Células CACO-2 , Canabinoides/análise , Proliferação de Células/efeitos dos fármacos , Farinha/análise , Células HT29 , Humanos , Fenóis , Extratos Vegetais/química , Óleos de Plantas/química , Sementes/química , Espectrometria de Massas em Tandem
9.
Plant Foods Hum Nutr ; 75(4): 487-494, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32671681

RESUMO

The ethno-pharmaceutical use of the edible fixed oil produced from lentisk (Pistacia lentiscus) berries covers a long tradition in several Mediterranean regions. Many of the health-promoting properties of lentisk berry oil (LBO) have been associated with the content of polar (poly)phenolic compounds. However, the polar fraction (methanol 80%, v/v) of LBO (LBO-pf) remains poorly and inadequately characterized. We assessed the phytochemical composition (fatty acids, phytosterols and polyphenols) of cold-pressed LBO produced in Cilento (Campania region, Italy) over four years of production (2015-2018). Main phenolic compounds present in LBO-pf were identified and semi-quantified combining ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) and HPLC with diode array detection. Phenolic compounds, also responsible for oil stability and antioxidant properties, are relatively abundant in LBO, compared to other edible oils. LBO-pf induced clear dose-dependent effects on the growth of HT-29 cell line derived from human colorectal adenocarcinoma, as evidenced by the cell cycle arrest. Our data support the health-promoting properties of cold-pressed LBO, which is obtained with good yield from spontaneous plants growing in semiarid regions.


Assuntos
Pistacia , Proliferação de Células , Cromatografia Líquida de Alta Pressão , Frutas , Humanos , Itália , Compostos Fitoquímicos , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem
10.
J Proteome Res ; 17(7): 2412-2420, 2018 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-29851351

RESUMO

A long-term psychiatric 40 years-old male patient was found dead at 9:00 a.m. in the clinic where he lived. Death was caused by traumatic injuries, which the sanitary staff imputed to a fall. Nurses declared that the patient refused having breakfast, whereas at autopsy the stomach contained 350 g of whitish semifluid material. Using both shotgun and gel-based proteomics, we demonstrated that the chyme contained partly digested milk- and bread-derived proteins, eaten during a recent breakfast. The conflict between evidence and assertions of the attending sanitary staff prompted the Legal Authority to undertake detailed investigations to ascertain facts and possible responsibilities. The herein characterization provides insights in the in vivo mechanisms of gastric breakdown of food proteins in a real meal. ß-lactoglobulin was partially resistant to gastric digestion as confirmed by Western blot analysis, in contrast to caseins and wheat gluten proteins, which had been degraded by gastric fluids. In addition to a complex pattern of gastric proteins (e.g., mucin-5AC, pepsin A-3, pepsinogen C, gastric lipase, gastrokine-2, trefoil factors), chyme contained intact proteins and variably sized food-derived polypeptides arising from peptic and nonpeptic proteolytic cleavage as well as heterodimeric disulfide-cross-linked peptides. These findings suggest that the current analytical workflows offer only a partial picture of the real complexity of the human "digestome".


Assuntos
Autopsia/métodos , Ciências Forenses/métodos , Conteúdo Gastrointestinal/química , Proteômica/métodos , Adulto , Caseínas/metabolismo , Digestão , Glutens/metabolismo , Humanos , Masculino , Proteínas do Leite/metabolismo , Proteólise
11.
Molecules ; 24(1)2018 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-30591638

RESUMO

A series of chemical and biochemical parameters of edible hemp resources (seeds, oil, and flour) from the monoecious EU registered hemp genotype Fedora, was determined, including fatty acid profile, phytosterol composition, total phenolics, antioxidant activity, macro- and micro-elements. The fatty acid ω-3/ω-6 approached the nutritionally optimal 3/1 ratio. ß-sitosterol and other phytosterols sterols dominated the unsaponifiable fraction. Hemp seeds, flour, and oil contained 767 ± 41, 744 ± 29, and 21 ± 5 mg GAE kg-1 total polyphenols, respectively. The antioxidant potential of Fedora flour and seeds, evaluated through the DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay, was higher than that of oil. K and Mg were the most abundant macro-elements, particularly in flour, while the concentration of trace elements was Fe > Cu > Ni > Mn. The presence of an array of bioactive compound candidate Fedora products as health-promoting food matrices. The ATR-FTIR spectra of hemp-derived products indicated the proximate composition of macro-nutrients.


Assuntos
Cannabis/química , Farinha/análise , Óleos de Plantas/química , Plantas Comestíveis/química , Sementes/química , Antioxidantes/análise , Minerais/análise , Polifenóis/análise , Saponinas/análise , Espectroscopia de Infravermelho com Transformada de Fourier
12.
Biochim Biophys Acta ; 1864(10): 1402-10, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27451951

RESUMO

Nuclear aggregates of polyamines (NAPs) are supramolecular compounds generated by the self-assembly of protonated nuclear polyamines (spermine, spermidine and putrescine) and phosphate ions. In the presence of genomic DNA, the hierarchical process of self-structuring ultimately produces nanotube-like polymers that envelop the double helix. Because of their modular nature and their aggregation-disaggregation dynamics, NAPs confer plasticity and flexibility to DNA. Through the disposition of charges, NAPs also enable a bidirectional stream of information between the genome and interacting moieties. High mobility group (HMG) B1 is a non-histone chromosomal protein that binds to DNA and that influences multiple nuclear processes. Because genomic DNA binds to either NAPs or HMGB1 protein, we explored the ability of in vitro self-assembled NAPs (ivNAPs) to mediate the DNA-HMGB1 interaction. To this end, we structured DNA-NAPs-HMGB1 and DNA-HMGB1-NAPs ternary complexes in vitro through opportune sequential incubations. Mobility shift electrophoresis and atomic force microscopy showed that the DNA-ivNAPs-HGMB1 complex had conformational assets supposedly more suitable those of the DNA-HGMB1-ivNAPs to comply with the physiological and functional requirements of DNA. Our findings indicated that ivNAPs act as mediators of the DNA-HMGB1 interaction.


Assuntos
Núcleo Celular/metabolismo , DNA/metabolismo , Proteína HMGB1/metabolismo , Poliaminas/metabolismo , Agregados Proteicos/fisiologia , Genoma/genética , Humanos , Microscopia de Força Atômica/métodos , Conformação de Ácido Nucleico , Espermidina/metabolismo , Espermina/metabolismo
13.
Pediatr Allergy Immunol ; 28(3): 230-237, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-27992668

RESUMO

BACKGROUND: Extensively hydrolyzed casein formula (EHCF) has been proposed for the prevention and is commonly used for the treatment of cow's milk allergy (CMA). The addition of the probiotic Lactobacillus rhamnosus GG (LGG) to EHCF may induce faster acquisition of tolerance to cow's milk. The mechanisms underlying this effect are largely unexplored. We investigated the effects of EHCF alone or in combination with LGG on ß-lactoglobulin (BLG) sensitization in mice. METHODS: Three-week-old C3H/HeOuJ mice were sensitized by oral administration of BLG using cholera toxin as adjuvant at weekly intervals for 5 weeks (sensitization period). Two experimental phases were conducted: (i) EHCF or EHCF+LGG given daily, starting 2 weeks before the sensitization period and then given daily for 5 weeks and (ii) EHCF or EHCF+LGG given daily for 4 weeks, starting 1 week after the sensitization period. Diet free of cow's milk protein was used as control. Acute allergic skin response, anaphylactic symptom score, body temperature, intestinal permeability, anti-BLG serum IgE, and interleukin (IL)-4, IL-5, IL-10, IL-13, IFN-γ mRNA expression were analyzed. Peptide fractions of EHCF were characterized by reversed-phase (RP)-HPLC, MALDI-TOF mass spectrometry, and nano-HPLC/ESI-MS/MS. RESULTS: Extensively hydrolyzed casein formula administration before or after BLG-induced sensitization significantly reduced acute allergic skin reaction, anaphylactic symptom score, body temperature decrease, intestinal permeability increase, IL-4, IL-5, IL-13, and anti-BLG IgE production. EHCF increased expression of IFN-γ and IL-10. Many of these effects were significantly enhanced by LGG supplementation. The peptide panels were similar between the two study formulas and contained sequences that could have immunoregulatory activities. CONCLUSIONS: The data support dietary intervention with EHCF for CMA prevention and treatment through a favorable immunomodulatory action. The observed effects are significantly enhanced by LGG supplementation.


Assuntos
Caseínas/administração & dosagem , Lacticaseibacillus rhamnosus/imunologia , Lactoglobulinas/imunologia , Hipersensibilidade a Leite/terapia , Probióticos/uso terapêutico , Animais , Caseínas/imunologia , Bovinos , Cromatografia Líquida de Alta Pressão , Citocinas/metabolismo , Imunoglobulina E/sangue , Camundongos , Camundongos Endogâmicos C3H , Leite , Hipersensibilidade a Leite/imunologia , Reação em Cadeia da Polimerase em Tempo Real , Espectrometria de Massas em Tandem
14.
J Enzyme Inhib Med Chem ; 32(1): 672-682, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28274171

RESUMO

The proteome of liver biopsies from human obese (O) subjects has been compared to those of nonobese (NO) subjects using two-dimensional gel electrophoresis (2-DE). Differentially represented proteins were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS)-based peptide mass fingerprinting (PMF) and nanoflow-liquid chromatography coupled to electrospray-tandem mass spectrometry (nLC-ESI-MS/MS). Overall, 61 gene products common to all of the liver biopsies were identified within 65 spots, among which 25 ones were differently represented between O and NO subjects. In particular, over-representation of short-chain acyl-CoA dehydrogenase, Δ(3,5)-Δ(2,4)dienoyl-CoA isomerase, acetyl-CoA acetyltransferase, glyoxylate reductase/hydroxypyruvate reductase, fructose-biphosphate aldolase B, peroxiredoxin I, protein DJ-1, catalase, α- and ß-hemoglobin subunits, 3-mercaptopyruvate S-transferase, calreticulin, aminoacylase 1, phenazine biosynthesis-like domain-containing protein and a form of fatty acid-binding protein, together with downrepresentation of glutamate dehydrogenase, glutathione S-transferase A1, S-adenosylmethionine synthase 1A and a form of apolipoprotein A-I, was associated with the obesity condition. Some of these metabolic enzymes and antioxidant proteins have already been identified as putative diagnostic markers of liver dysfunction in animal models of steatosis or obesity, suggesting additional investigations on their role in these syndromes. Their differential representation in human liver was suggestive of their consideration as obesity human biomarkers and for the development of novel antiobesity drugs.


Assuntos
Fármacos Antiobesidade/farmacologia , Descoberta de Drogas , Fígado/metabolismo , Terapia de Alvo Molecular , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Proteínas/metabolismo , Adulto , Fármacos Antiobesidade/química , Biomarcadores/análise , Eletroforese em Gel Bidimensional , Feminino , Humanos , Fígado/efeitos dos fármacos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Proteínas/antagonistas & inibidores , Proteínas/química , Proteoma/efeitos dos fármacos
15.
World J Microbiol Biotechnol ; 33(10): 186, 2017 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-28956240

RESUMO

Lactococcus lactis subsp. lactis CRL 1584 isolated from a bullfrog hatchery produces a bacteriocin that inhibits both indigenous Citrobacter freundii (a Red-Leg Syndrome related pathogen) and Lactobacillus plantarum, and Listeria monocytogenes as well. Considering that probiotics requires high cell densities and/or bacteriocin concentrations, the effect of the temperature on L. lactis growth and bacteriocin production was evaluated to find the optimal conditions. Thus, the growth rate was maximal at 36 °C, whereas the highest biomass and bacteriocin activity was achieved between 20 and 30 °C and 20-25 °C, respectively. The bacteriocin synthesis was closely growth associated reaching the maximal values at the end of the exponential phase. Since bacteriocins co-production has been evidenced in bacterial genera, a purification of the bacteriocin/s from L. lactis culture supernatants was carried out. The active fraction was purified by cationic-exchange chromatography and then, a RP-HPLC was carried out. The purified sample was a peptide with a 3353.05 Da, a molecular mass that matches nisin Z, which turned out to be the only bacteriocin produced by L. lactis CRL 1584. Nisin Z showed bactericidal effect on C. freundii and L. monocytogenes, which increased in the presence L-lactic acid + H2O2. This is the first report on nisin Z production by L. lactis from a bullfrog hatchery that resulted active on a Gram-negative pathogen. This peptide has potential probiotic for raniculture and as food biopreservative for bullfrog meat.


Assuntos
Antibacterianos/biossíntese , Citrobacter freundii/efeitos dos fármacos , Lactococcus lactis/crescimento & desenvolvimento , Nisina/análogos & derivados , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/farmacologia , Cromatografia por Troca Iônica , Lactobacillus plantarum/efeitos dos fármacos , Lactococcus lactis/isolamento & purificação , Listeria monocytogenes/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Nisina/biossíntese , Nisina/farmacologia , Rana catesbeiana/microbiologia , Temperatura
16.
J Food Sci Technol ; 54(8): 2316-2323, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28740288

RESUMO

The aim of this work was to assess whether the characteristic polyphenol traits of cherry biotypes persisted in thermally processed cherry products, such as jam. Thus, the RP-HPLC-diode array detector profiles of both colorless polyphenols and anthocyanins from three cherry varieties (two sweet and one tart cherry) were compared with those of low-sugar jam sourced from the same cultivars. Individual components were characterized by mass spectrometry. The total phenolic and total anthocyanin content as well as the radical scavenging potential (residual 75-91, 88-91 and 73-75%, respectively) were only slightly reduced by deep thermal treatments. Apart from the interconversion among the isomers of chlorogenic acid, the profile of both colorless polyphenols and anthocyanins substantially survived the jam manufacturing under conventional temperature-time regimen (80 °C, 1 h). The species- and cultivar-specific polyphenol molecular asset, especially the anthocyanin pattern, has potential to be monitored for traceability purpose, aimed to the varietal assessment of cherry biotypes used for producing jam.

17.
Molecules ; 21(10)2016 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-27775667

RESUMO

In this study, a hydroalcoholic chestnut shell extract was characterized and tested on six different human cell lines. Gallic, ellagic, and syringic acids were the most abundant non-condensed compounds in the chestnut extract, as determined by high performance liquid chromatography (HPLC). Tannins were mainly represented by condensed monomeric units of epigallocatechin and catechin/epicatechin. After 48 h of treatment, only the human hepatoblastoma HepG2 cells reached an inhibition corresponding to IC50 with an increase of apoptosis and mitochondrial depolarization. The cytokinome evaluation before and after treatment revealed that the vascular endothelial growth factor (VEGF) and the tumor necrosis factor (TNF)-α decreased after the treatment, suggesting a potential anti-angiogenic and anti-inflammatory effect of this extract. Moreover, the metabolome evaluation by ¹H-NMR evidenced that the polyphenols extracted from chestnut shell (PECS) treatment affected the levels of some amino acids and other metabolites. Overall, these data highlight the effects of biomolecules on cell proliferation, apoptosis, cell cycle and mitochondrial depolarization, and on cytokinomics and metabolomics profiles.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Citocinas/metabolismo , Fagaceae/química , Metaboloma/efeitos dos fármacos , Polifenóis/farmacologia , Antineoplásicos Fitogênicos/química , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Ácido Elágico/química , Ácido Elágico/farmacologia , Ácido Gálico/análogos & derivados , Ácido Gálico/química , Ácido Gálico/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células HT29 , Células Hep G2 , Humanos , Células MCF-7 , Mitocôndrias/efeitos dos fármacos , Exsudatos de Plantas/química , Exsudatos de Plantas/farmacologia , Polifenóis/química , Taninos/química , Taninos/farmacologia
18.
Rapid Commun Mass Spectrom ; 28(5): 499-504, 2014 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-24497288

RESUMO

RATIONALE: In the nuclei of eukaryotic cells, polyamines and phosphate ions self-assemble via ionic interactions and hydrogen bonding, generating three families of supramolecular compounds that have been named large (l-), medium (m-) and small (s-) nuclear aggregates of polyamines (NAPs). In a simulated nuclear environment, polyamines and phosphate ions generate the in vitro NAPs (ivNAPs) that share strict structural and functional analogies with their cellular cognates. Mass spectrometric data are expected to provide important structural details of NAPs/ivNAPs. METHODS: We used both electrospray ionization (ESI) and nitrocellulose (NC) matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) to support a variety of analytical techniques previously addressed to structurally characterize NAPs/ivNAPs. RESULTS: The dominant m/z values of s-ivNAP (m/z 735, 749, 761) are compatible with a defined set of cyclic or linear aggregates. On the basis of the experimental molecular mass (a cluster centred at m/z 2980), the m-ivNAP corresponds to the supramolecular assembly of four modules of s-ivNAPs. No informative mass spectra were obtained for the l-ivNAP. CONCLUSIONS: MS data support the models of NAPs that have been inferred by using an array of analytical techniques. NC MALDI-MS contributed much more effectively than ESI-MS to the structural characterization of ivNAPs.


Assuntos
Poliaminas/análise , Poliaminas/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Colódio , Modelos Biológicos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
19.
Molecules ; 19(6): 8644-60, 2014 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-24962397

RESUMO

In this work, we characterized conjugated linolenic acids (e.g., punicic acid) as the major components of the hydrophilic fraction (80% aqueous methanol extract) from pomegranate (Punica granatum L.) seed oil (PSO) and evaluated their anti-inflammatory potential on some human colon (HT29 and HCT116), liver (HepG2 and Huh7), breast (MCF-7 and MDA-MB-231) and prostate (DU145) cancer lines. Our results demonstrated that punicic acid and its congeners induce a significant decrease of cell viability for two breast cell lines with a related increase of the cell cycle G0/G1 phase respect to untreated cells. Moreover, the evaluation of a great panel of cytokines expressed by MCF-7 and MDA-MB-231 cells showed that the levels of VEGF and nine pro-inflammatory cytokines (IL-2, IL-6, IL-12, IL-17, IP-10, MIP-1α, MIP-1ß, MCP-1 and TNF-α) decreased in a dose dependent way with increasing amounts of the hydrophilic extracts of PSO, supporting the evidence of an anti-inflammatory effect. Taken together, the data herein suggest a potential synergistic cytotoxic, anti-inflammatory and anti-oxidant role of the polar compounds from PSO.


Assuntos
Anti-Inflamatórios/farmacologia , Neoplasias da Mama/tratamento farmacológico , Inflamação/tratamento farmacológico , Ácidos Linolênicos/farmacologia , Lythraceae/metabolismo , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocinas/biossíntese , Citocinas/metabolismo , Sinergismo Farmacológico , Feminino , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Células HCT116 , Células HT29 , Células Hep G2 , Humanos , Interações Hidrofóbicas e Hidrofílicas , Células MCF-7 , Extratos Vegetais/farmacologia , Óleos de Plantas/metabolismo , Sementes/metabolismo
20.
Food Res Int ; 181: 114119, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38448102

RESUMO

Tara (Caesalpinia spinosa, Leguminosae) seed germ (TSG), a by-product of tara gum (E417) extraction, has been used as a protein- and polyphenol-rich food ingredient for human and animal nutrition. Nevertheless, TSG is the alleged culprit for a recent foodborne outbreak of even severe acute illnesses that have affected hundreds of individuals in the USA, perhaps triggered by nonprotein amino acids such as baikiain. Herein, the composition of TSG has been characterized at molecular level, with a focus on proteins, phenolics, lipids, and mineral composition. TSG contains 43.4 % (w/w) proteins, tentatively identified for the first time by proteomics, and 14 % lipids, consisting of 83.6 % unsaturated fatty acids, especially linoleic acid. Ash is surprising high (6.5 %) because of an elevated concentration of P, K, Ca, and Mg. The detection of a rare earth element such as gadolinium (Gd, 1.6 mg kg-1), likely sourced from anthropogenic pollution, suggests alternative hypotheses for the origin of TSG hazards.


Assuntos
Caesalpinia , Farinha , Animais , Humanos , Polifenóis , Sementes , Ácido Linoleico
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