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1.
Anal Bioanal Chem ; 411(28): 7595-7602, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31655856

RESUMO

Intestinal diamine oxidase (DAO) acts as a protective barrier against exogenous histamine. A deficit of DAO activity can lead to the appearance of histamine intolerance, a clinical condition that may be treated by a low-histamine diet and oral DAO supplementation to enhance intestinal histamine degradation. As sources of DAO, porcine kidneys and certain legume seedlings are suitable components for the formulation of a DAO supplement. The aim of this work was to develop a rapid and reliable methodology for the in vitro determination of DAO activity in food matrices based on an enzymatic assay coupled to UHPLC-FL. The proposed method showed a satisfactory linearity and sensitivity and provided a relative standard deviation lower than 3%, guaranteeing method precision, and a mean recovery greater than 99% both for lyophilized pea sprouts and porcine kidney protein extracts. A high specificity is a key attribute of this method due to the use of histamine as the reaction substrate and the direct quantification of its degradation. Moreover, the lack of interference of catalase and hydrogen peroxide is another advantage in comparison with previously published methods. Lyophilized pea sprouts showed the greatest histamine-degrading activity (0.40 ± 0.01 mU/mg), followed by porcine kidney protein extracts (0.23 ± 0.01 mU/mg) and commercial DAO supplements (0.09 ± 0.06 mU/mg). This technique could be used as a tool to validate the DAO activity of food matrices of potential interest for the treatment of histamine intolerance.


Assuntos
Amina Oxidase (contendo Cobre)/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Alimentos , Animais , Liofilização , Técnicas In Vitro , Rim/enzimologia , Pisum sativum/enzimologia , Reprodutibilidade dos Testes , Suínos
2.
Eur J Nutr ; 57(1): 339-349, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27730364

RESUMO

PURPOSE: Polyphenol metabolites are key mediators of the biological activities of polyphenols. This study aimed to evaluate the long-term effects of a high-fat high-sucrose (HFHS) diet on the metabolism of proanthocyanidins from grape seed extract (GSE). METHODS: Adult female Wistar-Kyoto rats were fed a standard (STD) or HFHS diet supplemented or not with GSE for 16 weeks. PA metabolites were determined by targeted HPLC-MS/MS analysis. RESULTS: A lower concentration of total microbial-derived PA metabolites was present in urine and the aqueous fraction of faeces in the HFHS + GSE group than in the STD + GSE group. In contrast, a tendency towards the formation of conjugated (epi)catechin metabolites in the HFHS + GSE group was observed. CONCLUSIONS: These results show that a HFHS diet significantly modifies PA metabolism, probably via: (1) a shift in microbial communities not counteracted by the polyphenols themselves; and (2) an up-regulation of hepatic enzymes.


Assuntos
Dieta Hiperlipídica/efeitos adversos , Sacarose Alimentar/administração & dosagem , Extrato de Sementes de Uva/química , Proantocianidinas/metabolismo , Vitis , Animais , Catequina/metabolismo , Dieta , Fezes/química , Feminino , Microbioma Gastrointestinal/fisiologia , Extrato de Sementes de Uva/administração & dosagem , Proantocianidinas/administração & dosagem , Proantocianidinas/urina , Ratos , Ratos Endogâmicos WKY
3.
Biochem J ; 473(11): 1523-36, 2016 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-27013146

RESUMO

Human RNase 6 is a cationic secreted protein that belongs to the RNase A superfamily. Its expression is induced in neutrophils and monocytes upon bacterial infection, suggesting a role in host defence. We present here the crystal structure of RNase 6 obtained at 1.72 Å (1 Å=0.1 nm) resolution, which is the first report for the protein 3D structure and thereby setting the basis for functional studies. The structure shows an overall kidney-shaped globular fold shared with the other known family members. Three sulfate anions bound to RNase 6 were found, interacting with residues at the main active site (His(15), His(122) and Gln(14)) and cationic surface-exposed residues (His(36), His(39), Arg(66) and His(67)). Kinetic characterization, together with prediction of protein-nucleotide complexes by molecular dynamics, was applied to analyse the RNase 6 substrate nitrogenous base and phosphate selectivity. Our results reveal that, although RNase 6 is a moderate catalyst in comparison with the pancreatic RNase type, its structure includes lineage-specific features that facilitate its activity towards polymeric nucleotide substrates. In particular, enzyme interactions at the substrate 5' end can provide an endonuclease-type cleavage pattern. Interestingly, the RNase 6 crystal structure revealed a novel secondary active site conformed by the His(36)-His(39) dyad that facilitates the polynucleotide substrate catalysis.


Assuntos
Ribonuclease Pancreático/química , Ribonuclease Pancreático/metabolismo , Sequência de Aminoácidos , Sítios de Ligação , Catálise , Cristalografia por Raios X , Humanos , Cinética , Dados de Sequência Molecular , Ligação Proteica , Estrutura Secundária de Proteína , Homologia de Sequência de Aminoácidos , Especificidade por Substrato
4.
J Vet Pharmacol Ther ; 40(1): 28-34, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27302387

RESUMO

Searching for new therapeutic options against septic arthritis in horses, this research was focused on the study of the kinetics and local side effects after the intra-articular treatment of horses with cefovecin sodium. A single dose (240 mg) of the drug (Convenia® ) was administered into the radiocarpal joint of adult healthy horses (n = 6), and drug concentrations in plasma and synovial fluid were determined by high-performance liquid chromatography (HPLC). Local tolerance was also studied based on the modification of different joint physiopathological parameters (pH, cellular, and protein concentration in synovial fluid). Although no clinically relevant joint damage was noticed, significant increases in the protein concentrations at 5 min and in the cellular concentration at 30 min after cefovecin administration were observed in the treated radiocarpal joints. The duration of the cefovecin above the minimal inhibitory concentration (MIC) ≤1 µg/mL was 28.80 ± 2.58 h in the radiocarpal joint and 16.00 ± 2.86 h in plasma. The results of this study showed that intra-articular administration of cefovecin sodium in horses could be considered in the future to manage septic arthritis in horses, as it offers a good pharmacokinetic behavior and good local tolerance.


Assuntos
Antibacterianos/farmacocinética , Cefalosporinas/farmacocinética , Animais , Antibacterianos/administração & dosagem , Antibacterianos/efeitos adversos , Artrite Infecciosa/tratamento farmacológico , Artrite Infecciosa/veterinária , Cefalosporinas/administração & dosagem , Cefalosporinas/efeitos adversos , Tolerância a Medicamentos , Feminino , Doenças dos Cavalos/tratamento farmacológico , Cavalos/metabolismo , Injeções Intra-Articulares/veterinária , Articulações/metabolismo , Masculino
5.
Aging Clin Exp Res ; 28(6): 1061-1065, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27380506

RESUMO

Scaling up and replication of successful innovative integrated care models for chronic diseases is one of the targets of the European Innovation Partnership on Active and Healthy Ageing (EIP on AHA). MACVIA-LR® (MAladies Chroniques pour un VIeillissement Actif en Languedoc-Roussillon) is a Reference Site of the EIP on AHA. The main objective of MACVIA-LR® is to develop innovative solutions in order to (1) improve the care of patients affected by chronic diseases, (2) reduce avoidable hospitalization and (3) scale up the innovation to regions of Europe. The MACVIA-LR® project also aims to assess all possible aspects of medicine-including non-pharmacologic approaches-in order to maintain health and prevent chronic diseases. These approaches include hydrotherapy and balneotherapy which can be of great importance if health promotion strategies are considered. Balneotherapy at Balaruc-les-Bains focusses on musculoskeletal diseases and chronic venous insufficiency of the lower limbs. Each year, over 46,000 people attend an 18-day course related to a new falls prevention initiative combining balneotherapy and education. On arrival, each person receives a flyer providing information on the risk of fall and, depending on this risk, a course is proposed combining education and physical activity. A pilot study assesses the impact of the course 6 and 12 months later. This health promotion strategy for active and healthy ageing follows the FEMTEC (World Federation of Hydrotherapy and Climatotherapy) concept.


Assuntos
Acidentes por Quedas/prevenção & controle , Envelhecimento/fisiologia , Balneologia/métodos , Doença Crônica , Promoção da Saúde , Doenças Musculoesqueléticas , Doença Crônica/epidemiologia , Doença Crônica/reabilitação , Europa (Continente)/epidemiologia , Exercício Físico/fisiologia , Promoção da Saúde/métodos , Promoção da Saúde/estatística & dados numéricos , Humanos , Doenças Musculoesqueléticas/epidemiologia , Doenças Musculoesqueléticas/reabilitação , Educação de Pacientes como Assunto
6.
Radiologia ; 58(2): 88-100, 2016.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-26860655

RESUMO

The study of the structures that make up the brachial plexus has benefited particularly from the high resolution images provided by 3T magnetic resonance scanners. The brachial plexus can have mononeuropathies or polyneuropathies. The mononeuropathies include traumatic injuries and trapping, such as occurs in thoracic outlet syndrome due to cervical ribs, prominent transverse apophyses, or tumors. The polyneuropathies include inflammatory processes, in particular chronic inflammatory demyelinating polyneuropathy, Parsonage-Turner syndrome, granulomatous diseases, and radiation neuropathy. Vascular processes affecting the brachial plexus include diabetic polyneuropathy and the vasculitides. This article reviews the anatomy of the brachial plexus and describes the technique for magnetic resonance neurography and the most common pathologic conditions that can affect the brachial plexus.


Assuntos
Plexo Braquial/diagnóstico por imagem , Imageamento por Ressonância Magnética , Plexo Braquial/anatomia & histologia , Humanos
7.
Acta Derm Venereol ; 95(4): 422-6, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25367888

RESUMO

All cases of MM diagnosed in 23 hospitals in Catalonia, from 2000 to 2007 were recorded and melanoma incidence calculated and adjusted for the European standard population via the direct method. The age standardised rate/100,000 inhabitants varied from 6.74 in 2000 to 8.64 in 2007 for all melanomas and from 4.79 to 5.80 for invasive MMs; the Breslow thickness was stable during the period. The increase in invasive melanoma incidence in the elderly was remarkable, the crude rate/100,000 inhabitants increasing from 11.04 (2000) to 15.49 (2007) in the 60-64 year population, while remaining more stable in the 30-34 year range, from 3.97 in 2000 to 4.55 in 2007, and with a tendency to decrease from 5.1 in 2000 to 2.5 in 2007 for the age range of 25-29 years. These lower age ranges are much more affected by immigration. Despite the large immigrant population (nearly one million immigrants arrived in Catalonia during the study period from countries with a low melanoma incidence), melanoma incidence in our region has risen considerably and this trend is likely to persist in the near future.


Assuntos
Melanoma/epidemiologia , Neoplasias Cutâneas/epidemiologia , Adulto , Distribuição por Idade , Idoso , Emigração e Imigração , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Sistema de Registros , Espanha/epidemiologia , Adulto Jovem
8.
PLoS Pathog ; 8(11): e1003005, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23133388

RESUMO

Antimicrobial proteins and peptides (AMPs) are important effectors of the innate immune system that play a vital role in the prevention of infections. Recent advances have highlighted the similarity between AMPs and amyloid proteins. Using the Eosinophil Cationic Protein as a model, we have rationalized the structure-activity relationships between amyloid aggregation and antimicrobial activity. Our results show how protein aggregation can induce bacteria agglutination and cell death. Using confocal and total internal reflection fluorescence microscopy we have tracked the formation in situ of protein amyloid-like aggregates at the bacteria surface and on membrane models. In both cases, fibrillar aggregates able to bind to amyloid diagnostic dyes were detected. Additionally, a single point mutation (Ile13 to Ala) can suppress the protein amyloid behavior, abolishing the agglutinating activity and impairing the antimicrobial action. The mutant is also defective in triggering both leakage and lipid vesicle aggregation. We conclude that ECP aggregation at the bacterial surface is essential for its cytotoxicity. Hence, we propose here a new prospective biological function for amyloid-like aggregates with potential biological relevance.


Assuntos
Amiloide/química , Antibacterianos/química , Bactérias/química , Proteína Catiônica de Eosinófilo/química , Imunidade Inata , Viabilidade Microbiana , Substituição de Aminoácidos , Amiloide/genética , Amiloide/imunologia , Antibacterianos/imunologia , Bactérias/genética , Bactérias/imunologia , Infecções Bacterianas , Proteína Catiônica de Eosinófilo/genética , Proteína Catiônica de Eosinófilo/imunologia , Humanos , Mutação de Sentido Incorreto
9.
Lipids Health Dis ; 13: 31, 2014 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-24512213

RESUMO

BACKGROUND: Marine polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been associated with improvement in the Metabolic Syndrome (MS). The aim of this study is to evaluate how three fish-oil diets with different eicosapentaenoic acid/docosahexaenoic acid ratios (EPA/DHA ratio) affect the histology of liver, kidney, adipose tissue and aorta in a preliminary morphological study. This work uses an animal model of metabolic syndrome in comparison with healthy animals in order to provide information about the best EPA:DHA ratio to prevent or to improve metabolic syndrome symptoms. METHODS: 35 Wistar rats, as a control, and 35 spontaneously hypertensive obese rats (SHROB) were fed for 13 weeks with 3 different supplementation of fish oil containing EPA and DHA ratios (1:1, 2:1 and 1:2, respectively). All samples were stained with haematoxylin/eosin stain, except aorta samples, which were stained also with Verhoeff and van Gieson's stain. A histological study was carried out to evaluate changes. These changes were statistically analyzed using SPSS IBM 19 software. The quantitative data were expressed by mean ± SD and were compared among groups and treatments using ANOVA with post-hoc tests for parametric data and the U-Mann-Whitney for non-parametric data. Qualitative data were expressed in frequencies, and compared with contingency tables using χ² statistics. RESULTS: EPA:DHA 1:1 treatment tended to improve the density and the wrinkling of elastic layers in SHROB rats. Only Wistar rats fed with EPA:DHA 1:1 treatment did not show mast cells in adipose tissue and has less kidney atrophy. In both strains EPA:DHA 1:1 treatment improved inflammation related parameters in liver and kidney. CONCLUSIONS: EPA:DHA 1:1 treatment was the most beneficial treatment since improved many histological parameters in both groups of rats.


Assuntos
Suplementos Nutricionais , Ácidos Docosa-Hexaenoicos/administração & dosagem , Ácido Eicosapentaenoico/administração & dosagem , Síndrome Metabólica/tratamento farmacológico , Obesidade/tratamento farmacológico , Gordura Abdominal/efeitos dos fármacos , Gordura Abdominal/patologia , Animais , Aorta/efeitos dos fármacos , Aorta/patologia , Ácidos Docosa-Hexaenoicos/farmacologia , Avaliação Pré-Clínica de Medicamentos , Ácido Eicosapentaenoico/farmacologia , Rim/efeitos dos fármacos , Rim/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Síndrome Metabólica/patologia , Obesidade/patologia , Ratos , Ratos Wistar
10.
Biochem J ; 456(1): 99-108, 2013 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-23962023

RESUMO

Vertebrate secreted RNases (ribonucleases) are small proteins that play important roles in RNA metabolism, angiogenesis or host defence. In the present study we describe the antimicrobial properties of the N-terminal domain of the hcRNases (human canonical RNases) and show that their antimicrobial activity is well conserved among their lineage. Furthermore, all domains display a similar antimicrobial mechanism, characterized by bacteria agglutination followed by membrane permeabilization. The results of the present study show that, for all antimicrobial hcRNases, (i) activity is retained at the N-terminus and (ii) the antimicrobial mechanism is conserved. Moreover, using computational analysis we show that antimicrobial propensity may be conserved at the N-terminus for all vertebrate RNases, thereby suggesting that a defence mechanism could be a primary function in vertebrate RNases and that the N-terminus was selected to ensure this property. In a broader context, from the overall comparison of the peptides' physicochemical and biological properties, general correlation rules could be drawn to assist in the structure-based development of antimicrobial agents.


Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Bactérias/efeitos dos fármacos , Ribonucleases/química , Aglutinação , Sequência de Aminoácidos , Animais , Peptídeos Catiônicos Antimicrobianos/síntese química , Peptídeos Catiônicos Antimicrobianos/química , Bactérias/imunologia , Sequência Conservada , Evolução Molecular , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/imunologia , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Positivas/imunologia , Hemólise , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Lipossomos/química , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Fosfatidilcolinas/química , Fosfatidilgliceróis/química , Filogenia , Ribonucleases/imunologia , Ribonucleases/fisiologia , Ovinos
11.
Nutrients ; 16(8)2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38674832

RESUMO

A retrospective pilot study was carried out to investigate the prevalence of four variants of the diamine oxidase (DAO) encoding gene (AOC1) in Caucasian adults with symptoms of histamine intolerance. In a cohort of 100 patients and 100 healthy individuals, DAO-encoding gene non-synonymous Single Nucleotide Variations (SNVs) were genotyped by multiplex single-nucleotide primer extension (SNPE) and capillary electrophoresis, and serum DAO activity was analyzed with a radio-extraction assay. The study found that 79% of individuals with symptoms of histamine intolerance harbored one or more of the four SNVs associated with reduced DAO activity. No significant differences were found in the prevalence of any variant between the group of patients and healthy controls. However, when considering the status of the alleles associated with DAO deficiency, more homozygous alleles were observed in histamine-intolerant patients. Moreover, a slightly but statistically higher percentage of patients had a high genetic risk score, reflecting the cumulative effect of carrying multiple DAO deficiency-associated gene variants and a high load of risk alleles (homozygous). A relationship between serum DAO activity and the genetic load of one specific SNV was observed, with DAO activity being significantly lower in patients homozygous for rs2052129. These results potentially support that carrying multiple DAO deficiency-associated gene variants and a high load of risk alleles (homozygous) is more relevant than the mere presence of one or more SNVs. Further studies are needed to determine the predictive value of these DAO-encoding gene variants.


Assuntos
Amina Oxidase (contendo Cobre) , Histamina , Polimorfismo de Nucleotídeo Único , Humanos , Amina Oxidase (contendo Cobre)/genética , Amina Oxidase (contendo Cobre)/sangue , Projetos Piloto , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , Histamina/sangue , Estudos Retrospectivos , Alelos , Prevalência , Idoso , Predisposição Genética para Doença , Genótipo , População Branca/genética
12.
Antimicrob Agents Chemother ; 57(8): 3797-805, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23716047

RESUMO

There is an urgent need to develop new agents against mycobacterial infections, such as tuberculosis and other respiratory tract or skin affections. In this study, we have tested two human antimicrobial RNases against mycobacteria. RNase 3, also called the eosinophil cationic protein, and RNase 7 are two small cationic proteins secreted by innate cells during host defense. Both proteins are induced upon infection displaying a wide range of antipathogen activities. In particular, they are released by leukocytes and epithelial cells, contributing to tissue protection. Here, the two RNases have been proven effective against Mycobacterium vaccae at a low micromolar level. High bactericidal activity correlated with their bacterial membrane depolarization and permeabilization activities. Further analysis on both protein-derived peptides identified for RNase 3 an N-terminus fragment that is even more active than the parental protein. Also, a potent bacterial agglutinating activity was unique to RNase 3 and its derived peptide. The particular biophysical properties of the RNase 3 active peptide are envisaged as a suitable reference for the development of novel antimycobacterial drugs. The results support the contribution of secreted RNases to the host immune response against mycobacteria.


Assuntos
Antibacterianos/farmacologia , Proteína Catiônica de Eosinófilo/farmacologia , Mycobacterium/efeitos dos fármacos , Ribonucleases/farmacologia , Sequência de Aminoácidos , Antibacterianos/síntese química , Membrana Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular , Ativação Enzimática , Proteína Catiônica de Eosinófilo/síntese química , Genes Sintéticos , Humanos , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Dados de Sequência Molecular , Proteínas Recombinantes/farmacologia , Ribonucleases/síntese química
13.
Bioinformatics ; 28(1): 130-1, 2012 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-22053077

RESUMO

SUMMARY: AMPA is a web application for assessing the antimicrobial domains of proteins, with a focus on the design on new antimicrobial drugs. The application provides fast discovery of antimicrobial patterns in proteins that can be used to develop new peptide-based drugs against pathogens. Results are shown in a user-friendly graphical interface and can be downloaded as raw data for later examination. AVAILABILITY: AMPA is freely available on the web at http://tcoffee.crg.cat/apps/ampa. The source code is also available in the web. CONTACT: marc.torrent@upf.edu; david.andreu@upf.edu SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.


Assuntos
Algoritmos , Anti-Infecciosos/química , Peptídeos/química , Peptídeos Catiônicos Antimicrobianos/química , Internet , Linguagens de Programação , Estrutura Terciária de Proteína , Software
14.
Lipids Health Dis ; 12: 140, 2013 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-24083393

RESUMO

BACKGROUND: High consumption of fish carries a lower risk of cardiovascular disease as a consequence of dietary omega-3 long chain polyunsaturated fatty acid (n-3 PUFA; especially EPA and DHA) content. A controversy exists about the component/s responsible of these beneficial effects and, in consequence, which is the best proportion between both fatty acids. We sought to determine, in healthy Wistar rats, the proportions of EPA and DHA that would induce beneficial effects on biomarkers of oxidative stress, and cardiovascular disease risk. METHODS: Female Wistar rats were fed for 13 weeks with 5 different dietary supplements of oils; 3 derived from fish (EPA/DHA ratios of 1:1, 2:1, 1:2) plus soybean and linseed as controls. The activities of major antioxidant enzymes (SOD, CAT, GPX, and GR) were determined in erythrocytes and liver, and the ORAC test was used to determine the antioxidant capacity in plasma. Also measured were: C reactive protein (CRP), endothelial dysfunction (sVCAM and sICAM), prothrombotic activity (PAI-1), lipid profile (triglycerides, cholesterol, HDLc, LDLc, Apo-A1, and Apo-B100), glycated haemoglobin and lipid peroxidation (LDL-ox and MDA values). RESULTS: After three months of nutritional intervention, we observed statistically significant differences in the ApoB100/ApoA1 ratio, glycated haemoglobin, VCAM-1, SOD and GPx in erythrocytes, ORAC values and LDL-ox. Supplementation with fish oil derived omega-3 PUFA increased VCAM-1, LDL-ox and plasma antioxidant capacity (ORAC). Conversely, the ApoB100/ApoA1 ratio and percentage glycated haemoglobin decreased. CONCLUSIONS: Our results showed that a diet of a 1:1 ratio of EPA/DHA improved many of the oxidative stress parameters (SOD and GPx in erythrocytes), plasma antioxidant capacity (ORAC) and cardiovascular risk factors (glycated haemoglobin) relative to the other diets.


Assuntos
Doenças Cardiovasculares/prevenção & controle , Ácidos Docosa-Hexaenoicos/administração & dosagem , Ácido Eicosapentaenoico/administração & dosagem , Eritrócitos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Animais , Apolipoproteínas/sangue , Proteína C-Reativa/metabolismo , Catalase/metabolismo , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Dieta , Eritrócitos/metabolismo , Feminino , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Hemoglobinas Glicadas/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Lipoproteínas LDL/sangue , Fígado/metabolismo , Malondialdeído/metabolismo , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismo , Triglicerídeos/sangue
15.
Nutrients ; 15(12)2023 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-37375711

RESUMO

Vegetarian and vegan diets are increasingly being adopted in Spain, a trend mainly driven by ethical concerns for animal welfare and the environment. This has resulted in a growing market for plant-based substitutes of meat products. However, available data on the nutritional value of such meat analogues in Mediterranean countries are still limited. In this study, the labelling information of four categories of plant-based meat analogues (n = 100) and the corresponding conventional meat products (n = 48) available on the Spanish market was surveyed and compared. The nutrient content of plant-based meat analogues varied significantly, due to the wide range of ingredients used in their formulation. Some of these products were found to have a low protein content, which in others was enhanced by the addition of cereals and legumes. Compared to the meat products, the plant-based analogues contained lower levels of total fat as well as saturated fat, which ranged from 30% of total fat in burgers to less than 15% in meatballs, sausages, and nuggets; in contrast, they contained higher amounts of fiber and complex carbohydrates. Overall, the meat analogues cannot be considered as nutritionally equivalent substitutes to conventional meat products due to a high variability of protein content and other nutrients.


Assuntos
Produtos da Carne , Animais , Humanos , Carne , Valor Nutritivo , Dieta Vegana , Vegetarianos
16.
J Struct Biol ; 179(1): 1-9, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22579681

RESUMO

The human eosinophil cationic protein (ECP), also known as RNase 3, is an eosinophil secretion protein that is involved in innate immunity and displays antipathogen and proinflammatory activities. ECP has a high binding affinity for heterosaccharides, such as bacterial lipopolysaccharides and heparan sulfate found in the glycocalix of eukaryotic cells. We have crystallized ECP in complex with sulfate anions in a new monoclinic crystal form. In this form, the active site groove is exposed, providing an alternative model for ligand binding studies. By exploring the protein-sulfate complex, we have defined the sulfate binding site architecture. Three main sites (S1-S3) are located in the protein active site; S1 and S2 overlap with the phosphate binding sites involved in RNase nucleotide recognition. A new site (S3) that is unique to ECP is one of the key anchoring points for sulfated ligands. Arg 1 and Arg 7 in S3, together with Arg 34 and Arg 36 in S1, form the main basic clusters that assist in the recognition of ligand anionic groups. The location of additional sulfate bound molecules, some of which contribute to the crystal packing, may mimic the binding to extended anionic polymers. In conclusion, the structural data define a binding pattern for the recognition of sulfated molecules that can modulate the role of ECP in innate immunity. The results reveal the structural basis for the high affinity of ECP for glycosaminoglycans and can assist in structure-based drug design of inhibitors of the protein cytotoxicity to host tissues during inflammation.


Assuntos
Proteína Catiônica de Eosinófilo/química , Sulfatos/química , Sequência de Aminoácidos , Domínio Catalítico , Cristalização , Proteína Catiônica de Eosinófilo/metabolismo , Glicosaminoglicanos/química , Glicosaminoglicanos/metabolismo , Humanos , Ligantes , Simulação de Dinâmica Molecular , Dados de Sequência Molecular , Ligação Proteica , Conformação Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo
17.
Antimicrob Agents Chemother ; 56(5): 2378-85, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22330910

RESUMO

Antimicrobial proteins and peptides (AMPs) are essential effectors of innate immunity, acting as a first line of defense against bacterial infections. Many AMPs exhibit high affinity for cell wall structures such as lipopolysaccharide (LPS), a potent endotoxin able to induce sepsis. Hence, understanding how AMPs can interact with and neutralize LPS endotoxin is of special relevance for human health. Eosinophil cationic protein (ECP) is an eosinophil secreted protein with high activity against both Gram-negative and Gram-positive bacteria. ECP has a remarkable affinity for LPS and a distinctive agglutinating activity. By using a battery of LPS-truncated E. coli mutant strains, we demonstrate that the polysaccharide moiety of LPS is essential for ECP-mediated bacterial agglutination, thereby modulating its antimicrobial action. The mechanism of action of ECP at the bacterial surface is drastically affected by the LPS structure and in particular by its polysaccharide moiety. We have also analyzed an N-terminal fragment that retains the whole protein activity and displays similar cell agglutination behavior. Conversely, a fragment with further minimization of the antimicrobial domain, though retaining the antimicrobial capacity, significantly loses its agglutinating activity, exhibiting a different mechanism of action which is not dependent on the LPS composition. The results highlight the correlation between the protein's antimicrobial activity and its ability to interact with the LPS outer layer and promote bacterial agglutination.


Assuntos
Parede Celular/química , Proteína Catiônica de Eosinófilo/farmacologia , Escherichia coli/efeitos dos fármacos , Lipopolissacarídeos/química , Oligopeptídeos/farmacologia , Aglutinação/efeitos dos fármacos , Aderência Bacteriana/efeitos dos fármacos , Sítios de Ligação , Configuração de Carboidratos , Proteína Catiônica de Eosinófilo/química , Proteína Catiônica de Eosinófilo/metabolismo , Eosinófilos/citologia , Eosinófilos/imunologia , Eosinófilos/metabolismo , Escherichia coli/genética , Escherichia coli/fisiologia , Humanos , Lipopolissacarídeos/metabolismo , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Modelos Moleculares , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Relação Estrutura-Atividade
18.
Biol Chem ; 393(8): 801-15, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22944682

RESUMO

Antimicrobial RNases are small cationic proteins belonging to the vertebrate RNase A superfamily and endowed with a wide range of antipathogen activities. Vertebrate RNases, while sharing the active site architecture, are found to display a variety of noncatalytical biological properties, providing an excellent example of multitask proteins. The antibacterial activity of distant related RNases suggested that the family evolved from an ancestral host-defence function. The review provides a structural insight into antimicrobial RNases, taking as a reference the human RNase 3, also named eosinophil cationic protein (ECP). A particular high binding affinity against bacterial wall structures mediates the protein action. In particular, the interaction with the lipopolysaccharides at the Gram-negative outer membrane correlates with the protein antimicrobial and specific cell agglutinating activity. Although a direct mechanical action at the bacteria wall seems to be sufficient to trigger bacterial death, a potential intracellular target cannot be discarded. Indeed, the cationic clusters at the protein surface may serve both to interact with nucleic acids and cell surface heterosaccharides. Sequence determinants for ECP activity were screened by prediction tools, proteolysis and peptide synthesis. Docking results are complementing the structural analysis to delineate the protein anchoring sites for anionic targets of biological significance.


Assuntos
Antibacterianos/química , Proteína Catiônica de Eosinófilo/química , Fatores Imunológicos/química , Sequência de Aminoácidos , Animais , Antibacterianos/imunologia , Fenômenos Fisiológicos Bacterianos , Proteína Catiônica de Eosinófilo/imunologia , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Fatores Imunológicos/imunologia , Modelos Moleculares , Dados de Sequência Molecular , Alinhamento de Sequência
19.
Front Nutr ; 9: 973682, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36313101

RESUMO

Efforts are currently being directed to identify a non-invasive marker that can serve as a solid and clinically irrefutable diagnostic criterion for histamine intolerance associated with diamine oxidase (DAO) deficiency. Accordingly, the identification of biomarkers of histamine (HA) metabolism in urine is proposed as a possible new diagnostic strategy. It is hypothesized that individuals with histamine intolerance could have a different urinary profile of HA and its metabolites in comparison with the healthy population. Thus, the aim of this study was to assess the urinary excretion of HA and 1-methylhistamine (MHA) in individuals diagnosed with histamine intolerance and in a control group. Levels of HA and MHA were compared between 24 h and first morning spot urine in a subgroup of 14 control individuals. Then, HA and MHA concentrations in spot urine of 32 histamine intolerant and 55 control individuals were determined by ultra-high performance liquid chromatography and fluorometric detection (UHPLC-FL) and normalized by creatinine. No differences were found between HA and MHA levels in 24 h and first morning samples. Overall, histamine intolerant patients presented a distinct urinary excretion profile compared to the control group due to lower levels of MHA. No differences in urinary MHA were observed related to serum DAO activity. Spot urine samples were thus validated as a reliable tool to determine the urinary excretion of HA and MHA. These results constitute a starting point for the study of HA metabolomics as a suitable and non-invasive approach to histamine intolerance diagnosis.

20.
Front Nutr ; 9: 1018463, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36337620

RESUMO

Restrictive diets for the treatment of different gastrointestinal disorders are reported to change the composition of intestinal microbiota. Recently, it has been proposed that individuals with histamine intolerance suffer from intestinal dysbiosis, having an overabundance of histamine-secreting bacteria, but how it is still unknown this state is affected by the usual dietary treatment of histamine intolerance [i.e., low-histamine diet and the supplementation with diamine oxidase (DAO) enzyme]. Thus, a preliminary study was carried out aiming to evaluate the potential changes on the composition of the intestinal microbiota in a group of five women diagnosed with histamine intolerance undergoing 9 months of the dietary treatment of histamine intolerance. After sequencing bacterial 16S rRNA genes (V3-V4 region) and analyzing the data using the EzBioCloud Database, we observed a reduction in certain histamine-secreting bacteria, including the genera Proteus and Raoultella and the specie Proteus mirabilis. Moreover, it was also observed an increase in Roseburia spp., a bacterial group frequently related to gut health. These changes could help to explain the clinical improvement experienced by histamine intolerant women underwent a dietary treatment.

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