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1.
J Food Sci ; 84(11): 3083-3090, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31599975

RESUMO

Studies on the discovery and function of antioxidants are consistently being performed because oxidative stress can cause various diseases. Many compounds and natural products have antioxidant activity in vitro; however, it is often difficult to reproduce their effects in vivo. Additionally, methods to measure antioxidant activities in cells are also scarce. Here, we investigated the antioxidant activity of milk proteins by observing the formation of arsenite-induced stress granules as a tool to evaluate antioxidant activity in cells. Milk proteins not only decreased the formation of stress granules in several cell types but also scavenged 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cations in vitro. In addition, milk proteins inhibited cellular senescence based on an SA-ß-galactosidase assay, and increased differentiation to myotubes from myoblasts isolated from the skeletal muscles of mouse pups. Taken together, our results demonstrate that milk proteins have an antiaging effect, especially prevention of skeletal muscle loss, through their antioxidant activities. PRACTICAL APPLICATION: Our results provide that antioxidant effects of milk proteins containing α-caseins, ß-caseins, and ß-lactoglobulin can mitigate aging-related damage induced by oxidative stress through showing inhibition of cellular senescence and increase of differentiation and maturation of myoblast. Therefore, we suggest that milk proteins could be potent health supplements to prevent aging-associated diseases, especially sarcopenia.


Assuntos
Antioxidantes/farmacologia , Caseínas/farmacologia , Bovinos , Lactoglobulinas/farmacologia , Leite/química , Envelhecimento/efeitos dos fármacos , Animais , Antioxidantes/análise , Arsenitos/farmacologia , Senescência Celular/efeitos dos fármacos , Feminino , Camundongos , Proteínas do Leite/farmacologia , Estresse Oxidativo/efeitos dos fármacos
2.
J Dairy Sci ; 102(11): 9598-9604, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31521365

RESUMO

Gastrointestinal conditions in which the transit of contents is altered may benefit from nutritional approaches to influencing health outcomes. Milk proteins modulate the transit of contents along different regions, suggesting that they have varying effects on neuromuscular function to alter gastrointestinal motility. We tested the hypothesis that bovine whey and casein milk protein hydrolysates could have direct modulatory effects on colonic motility patterns in isolated rat large intestine. Casein protein hydrolysate (CPH), whey protein concentrate (WPC), whey protein hydrolysate (WPH), and a milk protein hydrolysate (MPH; a hydrolyzed blend of 60% whey to 40% casein) were compared for their effects on spontaneous contractile waves. These contractions propagate along the length of the isolated intact large intestine (22 cm) between the proximal colon and rectum and were detected by measuring activity at 4 locations. Milk proteins were perfused through the tissue bath, and differences in contraction amplitude and frequency were quantified relative to pretreatment controls. Propagation frequency was decreased by CPH, increased by MPH, and unaffected by intact whey proteins. The reduced motility with CPH and increased motility with MPH indicate a direct action of these milk proteins on colon tissue and provide evidence for differential modulation by hydrolysate type. These findings mirror actions on lower gastrointestinal transit reported in vivo, with the exception of WPH, suggesting that other factors are required.


Assuntos
Caseínas/farmacologia , Colo/efeitos dos fármacos , Trânsito Gastrointestinal , Contração Muscular/efeitos dos fármacos , Proteínas do Soro do Leite/farmacologia , Animais , Bovinos , Intestino Grosso , Masculino , Hidrolisados de Proteína/farmacologia , Ratos , Ratos Sprague-Dawley , Reprodução
3.
J Dairy Sci ; 102(12): 10711-10723, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31548055

RESUMO

The objective of this work was to obtain casein hydrolysates with aspartic proteinases present in extracts from the artichoke flower (Cynara scolymus L.) and evaluate their antioxidant, antimicrobial, and angiotensin-I converting enzyme (ACE) inhibitory activity in vitro. The casein hydrolysates produced by the action of C. scolymus had elevated antihypertensive and antioxidant activity due to their high hydrophobic peptide content (93.84, 96.58, and 90.54% at 2, 4, and 16 h of hydrolysis, respectively). Hydrolysis time and molecular weight (<3 kDa) had a significant influence on the hypertensive and antioxidant activity of the hydrolysates, which were greater at hydrolysis times of 4 and 16 h and corresponding to the <3 kDa fractions. The <3 kDa fraction of the 16 h hydrolysate had an ACE inhibitory activity with a half-maximal inhibitory concentration (IC50) of 71.77 µg peptides per mL; DPPH and ABTS•+ radical scavenging activities of 6.27 µM and 6.21 mM Trolox equivalents per mg of peptides, respectively; and iron (II) chelation activity with an IC50 of 221.49 µg of peptides per mL. Antimicrobial activity against Enterococcus faecalis was also observed in the hydrolysates. From the peptide sequences identified in the hydrolysates, we detected 22 peptides (from the BIOPEP database) that were already in their bioactive form (AMKPWIQPK, AMKPWIQPKTKVIPYVRYL, ARHPHPHLSFM, DAQSAPLRVY, FFVAPFPEVFGK, GPVRGPFPII, KVLPVPQK, LLYQEPVLGPVRGPFPIIV, MAIPPKKNQDK, NLHLPLPLL, PAAVRSPAQILQ, RELEELNVPGEIVESLSSSEESITR, RPKHPIKHQ, RPKHPIKHQGLPQEVLNENLLRF, SDIPNPIGSENSEK, TPVVVPPFLQP, VENLHLPLPLL, VKEAMAPK, VLNENLLR, VYPFPGPIH, VYQHQKAMKPWIQPKTKVIPYVRY, VYQHQKAMKPWIQPKTKVIPYVRYL) and are reported to display antioxidant, antimicrobial, and ACE inhibitory activity. We also identified 12,116, 14,513, and 25,169 peptide sequences in the hydrolysates at 2, 4, and 16 h, respectively, that were contained in the primary sequence, and these are reported to display ACE inhibitory, antioxidant, dipeptidyl peptidase IV inhibition, antithrombotic, opioid, immunomodulation, antiamnesic, anticancer, chelating, and hemolytic bioactivity.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Anti-Infecciosos/farmacologia , Antioxidantes/farmacologia , Caseínas/farmacologia , Cynara scolymus/enzimologia , Peptídeo Hidrolases/metabolismo , Hidrolisados de Proteína/farmacologia , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Animais , Anti-Infecciosos/isolamento & purificação , Anti-Hipertensivos/isolamento & purificação , Anti-Hipertensivos/farmacologia , Antioxidantes/química , Caseínas/isolamento & purificação , Bovinos , Peso Molecular , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação
4.
Molecules ; 24(16)2019 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-31408975

RESUMO

Intrinsically disordered proteins play a central role in dynamic regulatory and assembly processes in the cell. Recently, a human κ-casein proteolytic fragment called lactaptin (8.6 kDa) was found to induce apoptosis of human breast adenocarcinoma MCF-7 and MDA-MB-231 cells with no cytotoxic activity toward normal cells. Earlier, we had designed some recombinant analogs of lactaptin and compared their biological activity. Among these analogs, RL2 has the highest antitumor activity, but the amino acid residues and secondary structures that are responsible for RL2's activity remain unclear. To elucidate the structure-activity relations of RL2, we studied the structural and aggregation features of this fairly large intrinsically disordered fragment of human milk κ-casein by a combination of physicochemical methods: NMR, paramagnetic relaxation enhancement (PRE), Electron Paramagnetic Resonance (EPR), circular dichroism, dynamic light scattering, atomic force microscopy, and a cytotoxic activity assay. It was found that in solution, RL2 exists as stand-alone monomeric particles and large aggregates. Whereas the disulfide-bonded homodimer turned out to be more prone to assembly into large aggregates, the monomer predominantly forms single particles. NMR relaxation analysis of spin-labeled RL2 showed that the RL2 N-terminal region, which is essential not only for multimerization of the peptide but also for its proapoptotic action on cancer cells, is more ordered than its C-terminal counterpart and contains a site with a propensity for α-helical secondary structure.


Assuntos
Antineoplásicos/química , Caseínas/química , Peptídeos Penetradores de Células/química , Proteínas Intrinsicamente Desordenadas/química , Sequência de Aminoácidos , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Caseínas/biossíntese , Caseínas/genética , Caseínas/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Peptídeos Penetradores de Células/biossíntese , Peptídeos Penetradores de Células/genética , Peptídeos Penetradores de Células/farmacologia , Clonagem Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Humanos , Proteínas Intrinsicamente Desordenadas/biossíntese , Proteínas Intrinsicamente Desordenadas/genética , Proteínas Intrinsicamente Desordenadas/farmacologia , Células MCF-7 , Agregados Proteicos/genética , Multimerização Proteica , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Relação Estrutura-Atividade
5.
J Dairy Sci ; 102(10): 8604-8613, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31378502

RESUMO

The objective of this work was to better understand the effect of differences in milk protein composition, and specifically, a change in ß-casein to total casein in a milk-based matrix, on growth performance and metabolic and inflammatory responses using a piglet model. Three formulas were optimized for piglets, with similar metabolizable energy, total protein content, and other essential nutrients. Only the protein type and ratio varied between the treatments: the protein fraction of the control diet contained only whey proteins, whereas 2 other matrices contained a whey protein to casein ratio of 60:40, and differed in the amount of ß-casein (12.5 and 17.1% of total protein). Piglets fed formula containing whey proteins and caseins, regardless of the concentration of ß-casein, showed a significantly higher average daily gain, average daily feed intake, and feed efficiency compared with piglets consuming the formula with only whey protein. Consumption of the formula containing only whey protein showed higher levels of plasma glucagon-like peptide-1 and ghrelin compared with the consumption of formula containing casein and whey protein. A positive correlation was observed between postprandial time and glucagon-like peptide-1 response. The intestinal pro-inflammatory cytokine tumor necrosis factor α increased significantly in piglets fed the whey protein/casein diet compared with those fed whey protein formula. All formula-fed piglets showed a lower level of IL-6 cytokine compared with the ad libitum sow-fed piglets, regardless of composition. No significant differences in the anti-inflammatory IL-10 concentration were observed between treatment groups. Milk protein composition contributed to the regulation of piglets' metabolic and physiological responses, with whey protein/casein formula promoting growth performance and a different immune regulatory balance compared with a formula containing only whey protein. Results indicated no differences between treatments containing different levels of ß-casein.


Assuntos
Ração Animal , Caseínas/farmacologia , Citocinas/metabolismo , Proteínas do Leite/farmacologia , Animais , Caseínas/metabolismo , Dieta/veterinária , Metabolismo Energético , Feminino , Interleucina-10/metabolismo , Masculino , Leite/metabolismo , Proteínas do Leite/química , Proteínas do Leite/metabolismo , Distribuição Aleatória , Suínos , Proteínas do Soro do Leite/análise , Proteínas do Soro do Leite/metabolismo , Proteínas do Soro do Leite/farmacologia
6.
Biomed Res Int ; 2019: 4087160, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31317028

RESUMO

Autophagy is a degradative process in which cellular organelles and proteins are recycled to restore homeostasis and cellular metabolism. Autophagy can be either a prosurvival or a prodeath process and remains one of the most fundamental processes for cell vitality. Thus autophagy modulation is an important approach for reinforcement anticancer therapeutics. Earlier we have demonstrated that recombinant analog of human milk protein lactaptin (RL2) induced apoptosis of various cultured cancer cells and activated lipidation of microtubule-associated protein 1 light chain 3 (LC3). In this study we investigated whether autophagy inhibitors-chloroquine (CQ), Ku55933 (Ku), and 3-methyladenine (3MA)-or inducer-rapamycin (Rap)-can enhance cytotoxic activity of lactaptin analog in cancer cells and its anticancer activity in the mice model. Western Blot analysis revealed that RL2 induced short-term autophagy in MDA-MB-231 and MCF-7 cells at early stages of incubation and that these data were confirmed by the transmission electron microscopy of autophagosome/autophagolysosome formation. RL2 stimulates reactive oxygen species (ROS) production, autophagosomes accumulation, upregulation of ATG5 with processing of LC3I to LC3II, and downregulation of p62/sequestosome 1 (p62). We have shown that autophagy modulators, CQ, Ku, and Rap, synergistically increased cytotoxicity of RL2, and RL2 with CQ induced autophagic cell death. In addition, CQ, Ku, and Rap in combination with RL2 decreased activity of lysosomal protease Cathepsin D. More importantly, combining RL2 with CQ, we improved antitumor effect in mice. Detected synergistic cytotoxic effects of both types of autophagy regulators, inhibitors, and inducers with RL2 against cancer cells allow us to believe that these combinations can be a basis for the new anticancer approach. Finally, we suppose that CQ and Rap promoting of short-term RL2-induced autophagy interlinks with final autophagic cell death.


Assuntos
Antineoplásicos/farmacologia , Autofagia/efeitos dos fármacos , Caseínas/farmacologia , Neoplasias/tratamento farmacológico , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Apoptose/efeitos dos fármacos , Autofagia/genética , Caseínas/genética , Catepsina D/genética , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cloroquina/farmacologia , Humanos , Lisossomos/efeitos dos fármacos , Lisossomos/genética , Células MCF-7 , Camundongos , Proteínas Associadas aos Microtúbulos/genética , Morfolinas/farmacologia , Neoplasias/genética , Pironas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Proteína Sequestossoma-1/genética
7.
Amino Acids ; 51(8): 1209-1220, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31321559

RESUMO

Up to now, numerous peptides/hydrolysates derived from casein and whey protein have shown angiotensin-I-converting enzyme (ACE) inhibitory. In this research, quantum topological molecular similarity (QTMS) indices of amino acids were utilized in quantitative sequence-activity modeling (QSAM) to predict the activity of a set of milk-driven peptides with ACE inhibition. Since the derived peptides have not the same number of residues, we overcame this issue by auto cross covariance (ACC) methodology. Then, some QSAMs were built to predict the pIC50 value of ACE peptides derived from Bovine Casein and Whey. The model established an acceptable relationship between the selected variables and the pIC50 of the peptides. To estimate the performance of the developed models, casein and whey proteins from human, goat, bovine and sheep were virtually broken by trypsin and chymotrypsin enzymes and the ACE activity of the resultant virtual peptides were predicted and some new ACE peptides were proposed.


Assuntos
Aminoácidos/análise , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Caseínas/farmacologia , Leite/química , Fragmentos de Peptídeos/farmacologia , Peptidil Dipeptidase A/química , Proteínas do Soro do Leite/farmacologia , Inibidores da Enzima Conversora de Angiotensina/química , Animais , Caseínas/química , Bovinos , Cabras , Humanos , Hidrólise , Modelos Moleculares , Fragmentos de Peptídeos/química , Ovinos , Proteínas do Soro do Leite/química
8.
Nutrients ; 11(7)2019 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-31340476

RESUMO

Immunomodulatory nutraceuticals have garnered special attention due to their therapeutic potential for the amelioration of many chronic inflammatory conditions. Macrophages are key players in the induction, propagation and resolution of inflammation, actively contributing to the pathogenesis and resolution of inflammatory disorders. As such, this study aimed to investigate the possible therapeutic effects bovine casein derived nutraceuticals exert on macrophage immunological function. Initial studies demonstrated that sodium caseinate induced a M2-like macrophage phenotype that was attributed to the kappa-casein subunit. Kappa-casein primed macrophages acquired a M2-like phenotype that expressed CD206, CD54, OX40L, CD40 on the cell surface and gene expression of Arg-1, RELM-α and YM1, archetypical M2 markers. Macrophages stimulated with kappa-casein secreted significantly reduced TNF-α and IL-10 in response to TLR stimulation through a mechanism that targeted the nuclear factor-κB signal transduction pathway. Macrophage proteolytic processing of kappa-casein was required to elicit these suppressive effects, indicating that a fragment other than C-terminal fragment, glycomacropeptide, induced these modulatory effects. Kappa-casein treated macrophages also impaired T-cell responses. Given the powerful immuno-modulatory effects exhibited by kappa-casein and our understanding of immunopathology associated with inflammatory diseases, this fragment has the potential as an oral nutraceutical and therefore warrants further investigation.


Assuntos
Caseínas/farmacologia , Imunidade Inata/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Caseínas/metabolismo , Células Cultivadas , Técnicas de Cocultura , Humanos , Fatores Imunológicos/metabolismo , Mediadores da Inflamação/imunologia , Mediadores da Inflamação/metabolismo , Ativação Linfocitária , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos Endogâmicos C57BL , Fragmentos de Peptídeos/metabolismo , Fenótipo , Transdução de Sinais
9.
BMC Oral Health ; 19(1): 121, 2019 06 19.
Artigo em Inglês | MEDLINE | ID: mdl-31217005

RESUMO

BACKGROUND: This study aimed to assess the efficacy of a 980-nm diode and 10.6-µm CO2 laser accompanied by tricalcium phosphate-5% sodium fluoride (fTCP) and casein phosphopeptide amorphous calcium phosphate (CPP-ACP) for the remineralization of primary teeth. METHODS: In total, 117 extracted primary anterior teeth were randomly divided into eight experimental and one control group: (I) control (polished enamel), (II) fTCP varnish, (III) fTCP + diode laser, (IV) fTCP + CO2 laser, (V) CPP-ACP, (VI) CPP-ACP + diode laser, (VII) CPP-ACP + CO2 laser, (VIII) diode laser, and (IX) CO2 laser. The microhardness of 12 samples in each group and the enamel porosity of one sample in each group were assessed before and after demineralization and 28 days after remineralization. Data were analysed using two-way ANOVA. RESULTS: Significant differences existed in microhardness (P = 0.004) and percentage of remineralization (P < 0.001) after remineralization among the material groups such that the highest mean was noted in the CPP-ACP group. No significant difference was noted in microhardness (P = 0.052) or percentage of remineralization (P = 0.981) after remineralization among the laser groups. In all groups, porosities increased after demineralization and slightly decreased after remineralization; the greatest reduction in porosity of the material groups was noted in the fTCP group, and the CO2 group among the laser groups. The interaction effect of materials and lasers was not significant (P > 0.05). CONCLUSION: The highest microhardness was achieved after remineralization with CPP-ACP. The efficacy of the diode and CO2 lasers was the same. No synergistic effect was found between materials and lasers. TRIAL REGISTRATION: This is not a human subject research.


Assuntos
Cariostáticos/farmacologia , Caseínas/farmacologia , Esmalte Dentário/efeitos dos fármacos , Fluoretos/farmacologia , Lasers de Gás , Lasers Semicondutores , Remineralização Dentária/métodos , Dente Decíduo/efeitos dos fármacos , Criança , Pré-Escolar , Feminino , Humanos , Técnicas In Vitro , Masculino , Propriedades de Superfície/efeitos dos fármacos
10.
Inflamm Res ; 68(8): 715-722, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31168680

RESUMO

OBJECTIVE: Nutritional factors such as extensively hydrolyzed casein (eHC) have been proposed to exert anti-inflammatory activity and affect clinical outcomes such as tolerance development in cow's milk allergy. Granzyme B (GrB) induces apoptosis in target cells and also controls the inflammatory response. Whether eHC could affect the activity of granzyme B and play a role in GrB-mediated inflammatory responses in vitro was unknown. METHODS: The activity of GrB was measured using the substrate Ac-IEPD-pNA. Inflammatory responses were induced with GrB in HCT-8 and THP-1 cells, and pro-inflammatory cytokines were determined at the transcriptional and protein level. RESULTS: GrB could induce the expression of IL-1ß in HCT-8 cells, and IL-8 and MCP-1 in THP-1 cells, respectively. Interestingly, GrB acted synergistically on LPS-induced inflammation in HCT-8 cells and eHC reduced pro-inflammatory responses in both GrB and LPS-mediated inflammation. Further analyses revealed that eHC could inhibit the biological activities and cytotoxic activities of GrB and then could reduce GrB-mediated inflammatory response. CONCLUSION: The results from the current study suggest that anti-inflammatory activity of extensively hydrolyzed casein is, to a certain extent, mediated through modulation of granzyme B activity and responses.


Assuntos
Anti-Inflamatórios/farmacologia , Caseínas/farmacologia , Granzimas/metabolismo , Linhagem Celular , Citocinas/genética , Citocinas/metabolismo , Granzimas/genética , Humanos , Lipopolissacarídeos/farmacologia
11.
Acta Biochim Biophys Sin (Shanghai) ; 51(6): 638-644, 2019 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-31081016

RESUMO

The purpose of this study was to obtain an engineered Aspergillus niger strain with high glucoamylase activity by overexpressing the glucoamylase gene glaA and α-amylase gene amyA in A. niger CICC2462. Three recombinant strains containing a single copy of amyA (1A), containing two copies of amyA (2A), and coexpressing amyA and glaA (AG), respectively, were constructed. The transcript levels of amyA in 1A and 2A were increased by 2.95 folds and 3.09 folds, respectively. The levels of amyA and glaA in AG were increased by 1.21 folds and 2.86 folds, but the maximum extracellular glucoamylase activities did not differ significantly. In addition, after 1% casein phosphopeptides (CPPs) was added to the fermentation medium, the maximum extracellular glucoamylase activities for strains 1A, 2A, and AG were 35,200, 37,300, and 40,710 U/ml, respectively, which were significantly higher than that of the parental strain CICC2462 (28,250 U/ml), while CPPs alone had no effect on the parental strain CICC2462. We demonstrate that overexpression of amyA and glaA substantially increases the expression and secretion of glucoamylase in A. niger, and CPPs effectively improves the yield of glucoamylase in recombinant A. niger strains overexpressing amyA and glaA. The newly developed strains and culture methods may have extensive industrial applications.


Assuntos
Aspergillus niger/genética , Proteínas Fúngicas/genética , Glucana 1,4-alfa-Glucosidase/genética , alfa-Amilases/genética , Aspergillus niger/enzimologia , Aspergillus niger/metabolismo , Caseínas/metabolismo , Caseínas/farmacologia , Fermentação/efeitos dos fármacos , Proteínas Fúngicas/metabolismo , Regulação Enzimológica da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Engenharia Genética/métodos , Glucana 1,4-alfa-Glucosidase/metabolismo , Fosfopeptídeos/metabolismo , Fosfopeptídeos/farmacologia , alfa-Amilases/metabolismo
12.
Prep Biochem Biotechnol ; 49(7): 639-648, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31131710

RESUMO

There are many diseases linked to oxidative stress, including cancer. Importantly, endogenous antioxidants are insufficient to protect against this process. Peptides derived from food proteins produced by hydrolysis have been investigated as exogenous antioxidants. The present study aimed to identify novel peptides with antioxidant potential produced from egg and milk proteins hydrolysis with two new fungal proteases isolated from Eupenicillium javanicum and Myceliophthora thermophila. The degree of hydrolysis at several time points was calculated and correlated to DPPH scavenging and metal chelating assays, all hydrolysates presented antioxidant activity. Casein hydrolyzed by the M. thermophila protease showed the best antioxidant activity. The identified sequences showed that the proportions of amino acids that influence antioxidant activity support the antioxidant assay. Our data reveal the conditions necessary for the successful generation of antioxidant peptides using two novel fungal proteases. This opens a potential new avenue for the design and manufacture of antioxidant molecules.


Assuntos
Albuminas/química , Antioxidantes/química , Caseínas/química , Proteínas do Ovo/química , Peptídeos/química , Proteínas do Soro do Leite/química , Albuminas/farmacologia , Animais , Antioxidantes/farmacologia , Caseínas/farmacologia , Proteínas do Ovo/farmacologia , Eupenicillium/enzimologia , Peptídeo Hidrolases/química , Peptídeos/farmacologia , Proteólise , Sordariales/enzimologia , Proteínas do Soro do Leite/farmacologia
13.
Colloids Surf B Biointerfaces ; 181: 149-157, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31128515

RESUMO

Whey-derived alpha-casozepine bioactive peptide (YLGYLEQLLR) was associated with previously optimized guar-gum film-PLGA nanoparticles, aiming to increase both stability across gastrointestinal tract and permeability across absorptive epithelia. Oral films associated with nanoparticles (FNp) enhance buccal absorption along with protection of carried bioactive molecules that are swallowed, with inherent increase of bioavailability. None of developed formulations induced significant loss of cell viability. Permeability across both buccal and intestinal cell barriers was enhanced when alpha-casozepine was carried by FNp system, when compared with film and nanoparticles alone, in a simulated gastrointestinal tract environment. Moreover, differences in permeability profile across buccal and intestinal epithelia were in accordance with the slower erosion of PLGA nanoparticles in a media of neutral pH, resembling oral cavity conditions, and a faster erosion in acidic conditions, as occurs in stomach, as observed by a continuous analysis of nanoparticle morphology over 980 min by atomic force microscopy. Additionally, apparent permeability of alpha-casozepine across TR146 human buccal carcinoma cells and Caco-2/HT29-MTX co-culture, carried by FNp was indeed superior when compared with peptide loaded in PLGA nanoparticles and in films alone or with free peptide control solution. Both FNp and PLGA nanoparticles alone enhanced the permeability of relaxing peptide compared with guar-gum films alone. An increased tongue adhesion when PLGA nanoparticles were added to the guar-gum films was also observed. Developed formulations improved both buccal an intestinal absorption of carried bioactive molecules without compromising cell viability.


Assuntos
Caseínas/química , Sistemas de Liberação de Medicamentos , Nanopartículas/química , Fragmentos de Peptídeos/química , Ácido Poliglicólico/química , Administração Oral , Caseínas/administração & dosagem , Caseínas/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Relação Dose-Resposta a Droga , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/química , Humanos , Mitocôndrias/efeitos dos fármacos , Nanopartículas/administração & dosagem , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/farmacologia , Ácido Poliglicólico/administração & dosagem , Relação Estrutura-Atividade , Células Tumorais Cultivadas
14.
Nutrients ; 11(3)2019 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-30870995

RESUMO

Food-derived bioactive peptides are reported as beneficial and safe for human health. Glycomacropeptide (GMP) is a milk-protein-derived peptide that, in addition to its nutritional value, retains many biological properties and has therapeutic effects in several inflammatory disorders. GMP was shown under in vitro and in vivo conditions to exert a number of activities that regulate the physiology of important body systems, namely the gastrointestinal, endocrine, and immune systems. This review represents a comprehensive compilation summarizing the current knowledge and updated information on the major biological properties associated with GMP. GMP bioactivity is addressed with special attention on mechanisms of action, signaling pathways involved, and structural characteristics implicated. In addition, the results of various studies dealing with the effects of GMP on models of inflammatory diseases are reviewed and discussed.


Assuntos
Caseínas/administração & dosagem , Caseínas/farmacologia , Redes e Vias Metabólicas/efeitos dos fármacos , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/farmacologia , Caseínas/química , Análise de Alimentos , Humanos , Fragmentos de Peptídeos/química
15.
Microsc Res Tech ; 82(7): 1065-1072, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30884045

RESUMO

The objective of this in vitro study was to analyze and compare the biomimetic remineralizing efficacy of the self-assembling peptide (P11-4) with agents containing casein phoshopeptide-amorhous calcium phosphate fluoride (CPP-ACFP) and sodium fluoride (NaF) on artificial caries lesions using DIAGNOdent and micro-computed tomography (µCT). Artificial enamel lesions were prepared on extracted impacted sound mandibular third molars. The samples were randomly allocated to four groups (n = 8): Group 1, P11-4 (Curodont Repair, Credentis AG, Switzerland); Group 2, CPP-ACFP (MI Varnish, GCCo., Japan); Group3, NaF (Duraphat Varnish, Colgate, Colgate-Palmolive, NY, USA); Group 4, artificial saliva (control). The agents were applied to demineralized surfaces according to manufacturers' instructions; all specimens were stored in artificial saliva for 1 month. Demineralization and remineralization on enamel surfaces were analyzed and quantified by DIAGNOdent (KaVo, Germany) and µCT (SkyScan1174, Belgium) for lesion depth/area/volume/mineral density (MD). The remineralization efficacy of the agents was evaluated by DIAGNOdent on 1st, 7th, 30th days and by µCT on 30th day. Data were statistically analyzed by ANOVA, Kruskal-Wallis, T test, and Wilxocon tests. The highest remineralization efficacy findings in all periods were determined in Group 1, followed by Groups 2, 3, and 4. The remineralization findings for fluorescence, MD, lesion depth in Group 1 were found significantly higher (p < 0.01) than Group 3; and no significant differences (p > 0.05) were found between Groups 1-2 and Groups 2-3. The area and volume change values in Groups 1, 2, and 3 have shown no significancy (p > 0.05). A significant correlation (p < 0.01) was found between µCT and DIAGNOdent methods. The data of this study have demonstrated that P11-4 has showed the best remineralization efficacy, followed by CPP-ACFP and NaF. It is concluded that self-assembling peptide-based remineralization agent can be used successfully for biomimetic remineralization of enamel subsurface lesions.


Assuntos
Materiais Biomiméticos/farmacologia , Esmalte Dentário/efeitos dos fármacos , Esmalte Dentário/patologia , Peptídeos/farmacologia , Remineralização Dentária/métodos , Materiais Biomiméticos/química , Caseínas/farmacologia , Cárie Dentária , Fluoretos Tópicos , Humanos , Dente Molar/efeitos dos fármacos , Fluoreto de Sódio/farmacologia , Microtomografia por Raio-X
16.
Nutrients ; 11(3)2019 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-30823411

RESUMO

Introduction: In phenylketonuria (PKU), evidence suggests that casein glycomacropeptide supplemented with rate-limiting amino acids (CGMP-AA) is associated with better protein utilisation and less blood phenylalanine (Phe) variability. Aim: To study the impact of CGMP-AA on blood Phe variability using 3 different dietary regimens in children with PKU. Methods: This was a 6-week randomised controlled cross-over study comparing CGMP-AA vs. Phe-free l-amino acids (l-AA) assessing blood Phe and tyrosine (Tyr) variability over 24 h in 19 children (7 boys) with PKU, with a median age of 10 years (6⁻16). Subjects were randomised to 3 dietary regimens: (1) R1, CGMP-AA and usual dietary Phe (CGMP + Phe); (2) R2, CGMP-AA - Phe content of CGMP-AA from usual diet (CGMP - Phe); and (3) R3, l-AA and usual dietary Phe. Each regimen was administered for 14 days. Over the last 48 h on days 13 and 14, blood spots were collected every 4 h at 08 h, 12 h, 16 h, 20 h, 24 h, and 04 h. Isocaloric intake and the same meal plan and protein substitute dosage at standardised times were maintained when blood spots were collected. Results: Eighteen children completed the study. Median Phe concentrations over 24 h for each group were (range) R1, 290 (30⁻580), R2, 220 (10⁻670), R3, 165 (10⁻640) µmol/L. R1 vs. R2 and R1 vs. R3 p < 0.0001; R2 vs. R3 p = 0.0009. There was a significant difference in median Phe at each time point between R1 vs. R2, p = 0.0027 and R1 vs. R3, p < 0.0001, but not between any time points for R2 vs. R3. Tyr was significantly higher in both R1 and R2 [70 (20⁻240 µmol/L] compared to R3 [60 (10⁻200) µmol/L]. In children < 12 years, blood Phe remained in the target range (120⁻360 µmol/L), over 24 h, for 75% of the time in R1, 72% in R2 and 64% in R3; for children aged ≥ 12 years, blood Phe was in target range (120⁻600 µmol/L) in R1 and R2 for 100% of the time, but 64% in R3. Conclusions: The residual Phe in CGMP-AA increased blood Phe concentration in children. CGMP-AA appears to give less blood Phe variability compared to l-AA, but this effect may be masked by the increased blood Phe concentrations associated with its Phe contribution. Reducing dietary Phe intake to compensate for CGMP-AA Phe content may help.


Assuntos
Caseínas/administração & dosagem , Caseínas/farmacologia , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/farmacologia , Fenilalanina/sangue , Tirosina/sangue , Criança , Suplementos Nutricionais , Feminino , Humanos , Masculino , Fenilalanina/metabolismo , Fenilcetonúrias , Tirosina/metabolismo
17.
J Trace Elem Med Biol ; 52: 239-246, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30732889

RESUMO

Cadmium (Cd) exposure has been associated with an increased risk of cardiovascular diseases. The diet is a modifiable source of protecting or damaging factors that may affect this risk. Herein we tested the hypothesis that a soybean-based diet (SBD) protects the vascular wall of the aorta against Cd-induced pro-inflammatory and pro-apoptotic effects. To test this hypothesis, we fed male Wistar rats for 60 days with a casein-based diet (CBD) or an SBD. These animals were also exposed to tap-water without (CBD-Co/SBD-Co) or with 15(CBD-15Cd/SBD-15Cd) or 100 (CBD-100Cd/SBD-100Cd) ppm of Cd. Inflammatory parameters (mRNAs and/or proteins) were measured in thoracic aorta tissue. These included inducible and endothelial nitric oxide synthases, cyclooxygenase-2, intracellular-adhesion molecule-1, and vascular cell-adhesion molecule-1. As pro-apoptotic parameters, we measured Bax and Bcl-2 mRNA/protein, as well as TUNEL positive cells in the aorta tissue. Compared to CBD-Co, inflammatory and apoptosis markers increased in the aorta with the concentration of Cd in the drinking water. These effects were not observed in either SBD-15Cd or SBD-100Cd, which were similar to CBD-Co. Cd content in serum and in aortas from animals fed CBD-Co/SBD-15Cd or CBD-Co/SBD-100Cd were similar suggesting that, if any, the effect of SBD is not due to changes in Cd bioaccumulation, but due to secondary effects linked to the composition of the dietary soybean flour. Our findings are consistent with a protective effect of an SBD against Cd-induced inflammation and apoptosis in the thoracic aorta in a rat model.


Assuntos
Aorta Torácica/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Cádmio/toxicidade , Dieta , Inflamação/induzido quimicamente , Soja/química , Animais , Aorta Torácica/metabolismo , Aorta Torácica/patologia , Cádmio/administração & dosagem , Cádmio/análise , Caseínas/administração & dosagem , Caseínas/farmacologia , Inflamação/metabolismo , Inflamação/patologia , Masculino , Ratos , Ratos Wistar
18.
Lipids Health Dis ; 18(1): 41, 2019 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-30717735

RESUMO

Milk and dairy products are integral part of human nutrition and they are considered as the carriers of higher biological value proteins, calcium, essential fatty acids, amino acids, fat, water soluble vitamins and several bioactive compounds that are highly significant for several biochemical and physiological functions. In recent years, foods containing natural antioxidants are becoming popular all over the world as antioxidants can neutralize and scavenge the free radicals and their harmful effects, which are continuously produced in the biological body. Uncontrolled free radicals activity can lead to oxidative stresses, which have been implicated in breakdown of vital biochemical compounds such as lipids, protein, DNA which may lead to diabetes, accelerated ageing, carcinogenesis and cardiovascular diseases. Antioxidant capacity of milk and milk products is mainly due to sulfur containing amino acids, such as cysteine, phosphate, vitamins A, E, carotenoids, zinc, selenium, enzyme systems, superoxide dismutase, catalase, glutathione peroxidase, milk oligosaccharides and peptides that are produced during fermentation and cheese ripening. Antioxidant activity of milk and dairy products can be enhanced by phytochemicals supplementation while fermented dairy products have been reported contained higher antioxidant capacity as compared to the non-fermented dairy products. Literature review has shown that milk and dairy products have antioxidant capacity, however, information regarding the antioxidant capacity of milk and dairy products has not been previously compiled. This review briefly describes the nutritional and antioxidant capacity of milk and dairy products.


Assuntos
Antioxidantes/metabolismo , Laticínios , Leite , Aminoácidos/análise , Aminoácidos/metabolismo , Animais , Búfalos , Caseínas/metabolismo , Caseínas/farmacologia , Bovinos , Cabras , Humanos , Leite/química , Leite/metabolismo , Valor Nutritivo , Ovinos , Proteínas do Soro do Leite/metabolismo , Proteínas do Soro do Leite/farmacologia
19.
Eur J Appl Physiol ; 119(3): 697-711, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30610443

RESUMO

PURPOSE: Multiday racing causes mild left ventricular (LV) dysfunction from day 1 that persists on successive days. We evaluated ingesting casein protein hydrolysate-carbohydrate (PRO) compared with carbohydrate-only (CHO) during a 3-day mountain bike race. METHODS: Eighteen male cyclists were randomly assigned to ingest 6.7% carbohydrate without (CHO) or with 1.3% casein hydrolysate (PRO) during racing (~ 4-5 h/day; 68/71/71 km). Conventional LV echocardiography, plasma albumin content, plasma volume (PV) and blood biomarkers were measured before day 1 and post race on day 3. RESULTS: Fourteen cyclists (n = 7 per group) completed the race. PV increased in CHO (mean increase (95% CI), 10.2% (0.1 to 20.2)%, p = 0.045) but not in PRO (0.4% (- 6.1 to 6.9)%). Early diastolic transmitral blood flow (E) was unchanged but deceleration time from peak E increased post race (CHO: 46.7 (11.8 to 81.6) ms, p = 0.019; PRO: 24.2 (- 0.5 to 48.9) ms, p = 0.054), suggesting impaired LV relaxation. Tissue Doppler mitral annular velocity was unchanged in CHO, but in PRO septal early-to-late diastolic ratio decreased (p = 0.016) and was compensated by increased lateral early (p = 0.034) and late (p = 0.012) velocities. Systolic function was preserved in both groups; with increased systolic lateral wall velocity in PRO (p = 0.002). Effect size increase in serum creatine kinase (CK) activity, CK-MB and C-reactive protein concentrations was less in PRO than CHO (Cohen's d mean ± SD, PRO: 2.91 ± 2.07; CHO: 7.56 ± 4.81, p = 0.046). CONCLUSION: Ingesting casein hydrolysate with carbohydrate during a 3-day race prevented secondary hypervolemia and failed to curb impaired LV relaxation despite reducing tissue damage and inflammatory biomarkers. Without PV expansion, systolic function was preserved by lateral wall compensating for septal wall dysfunction.


Assuntos
Velocidade do Fluxo Sanguíneo/efeitos dos fármacos , Carboidratos/farmacologia , Caseínas/farmacologia , Função Ventricular Esquerda/efeitos dos fármacos , Biomarcadores/sangue , Ecocardiografia Doppler/métodos , Feminino , Humanos , Masculino , Hidrolisados de Proteína/efeitos dos fármacos , Disfunção Ventricular Esquerda/fisiopatologia
20.
Ecol Food Nutr ; 58(2): 80-92, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30650991

RESUMO

This study evaluated an alternative ocean-based fish protein, Advanced Protein Powder (APP) as a feasible, environmentally sustainable protein source to reduce childhood malnutrition. We completed a rodent feeding study to evaluate growth and development in young growing mice on a purified diet containing APP as compared to mice-fed diets using other common protein sources - casein, whey, and soy. Results suggested APP to be an effective and safe protein source and ensured normal body growth, bone development, and brain function in APP diet-fed mice. Evidence provided in this study supports considering the use of APP to reduce malnutrition among children worldwide.


Assuntos
Densidade Óssea/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Cognição/efeitos dos fármacos , Dieta , Proteínas na Dieta/farmacologia , Proteínas de Peixes/farmacologia , Ganho de Peso/efeitos dos fármacos , Animais , Encéfalo/efeitos dos fármacos , Caseínas/farmacologia , Caseínas/uso terapêutico , Criança , Proteínas na Dieta/uso terapêutico , Proteínas de Peixes/uso terapêutico , Peixes , Humanos , Masculino , Memória/efeitos dos fármacos , Camundongos , Oceanos e Mares , Desnutrição Proteico-Calórica/prevenção & controle , Proteínas de Soja/farmacologia , Proteínas de Soja/uso terapêutico , Proteínas do Soro do Leite/farmacologia , Proteínas do Soro do Leite/uso terapêutico
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