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1.
Anal Bioanal Chem ; 411(28): 7595-7602, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31655856

RESUMEN

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.


Asunto(s)
Amina Oxidasa (conteniendo Cobre)/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Alimentos , Animales , Liofilización , Técnicas In Vitro , Riñón/enzimología , Pisum sativum/enzimología , Reproducibilidad de los Resultados , Porcinos
2.
Eur J Nutr ; 57(1): 339-349, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27730364

RESUMEN

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.


Asunto(s)
Dieta Alta en Grasa/efectos adversos , Sacarosa en la Dieta/administración & dosificación , Extracto de Semillas de Uva/química , Proantocianidinas/metabolismo , Vitis , Animales , Catequina/metabolismo , Dieta , Heces/química , Femenino , Microbioma Gastrointestinal/fisiología , Extracto de Semillas de Uva/administración & dosificación , Proantocianidinas/administración & dosificación , Proantocianidinas/orina , Ratas , Ratas Endogámicas WKY
3.
Biochem J ; 473(11): 1523-36, 2016 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-27013146

RESUMEN

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.


Asunto(s)
Ribonucleasa Pancreática/química , Ribonucleasa Pancreática/metabolismo , Secuencia de Aminoácidos , Sitios de Unión , Catálisis , Cristalografía por Rayos X , Humanos , Cinética , Datos de Secuencia Molecular , Unión Proteica , Estructura Secundaria de Proteína , Homología de Secuencia de Aminoácido , Especificidad por Sustrato
4.
J Vet Pharmacol Ther ; 40(1): 28-34, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27302387

RESUMEN

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.


Asunto(s)
Antibacterianos/farmacocinética , Cefalosporinas/farmacocinética , Animales , Antibacterianos/administración & dosificación , Antibacterianos/efectos adversos , Artritis Infecciosa/tratamiento farmacológico , Artritis Infecciosa/veterinaria , Cefalosporinas/administración & dosificación , Cefalosporinas/efectos adversos , Tolerancia a Medicamentos , Femenino , Enfermedades de los Caballos/tratamiento farmacológico , Caballos/metabolismo , Inyecciones Intraarticulares/veterinaria , Articulaciones/metabolismo , Masculino
5.
Aging Clin Exp Res ; 28(6): 1061-1065, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27380506

RESUMEN

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.


Asunto(s)
Accidentes por Caídas/prevención & control , Envejecimiento/fisiología , Balneología/métodos , Enfermedad Crónica , Promoción de la Salud , Enfermedades Musculoesqueléticas , Enfermedad Crónica/epidemiología , Enfermedad Crónica/rehabilitación , Europa (Continente)/epidemiología , Ejercicio Físico/fisiología , Promoción de la Salud/métodos , Promoción de la Salud/estadística & datos numéricos , Humanos , Enfermedades Musculoesqueléticas/epidemiología , Enfermedades Musculoesqueléticas/rehabilitación , Educación del Paciente como Asunto
6.
Radiologia ; 58(2): 88-100, 2016.
Artículo en Inglés, Español | MEDLINE | ID: mdl-26860655

RESUMEN

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.


Asunto(s)
Plexo Braquial/diagnóstico por imagen , Imagen por Resonancia Magnética , Plexo Braquial/anatomía & histología , Humanos
7.
Acta Derm Venereol ; 95(4): 422-6, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25367888

RESUMEN

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.


Asunto(s)
Melanoma/epidemiología , Neoplasias Cutáneas/epidemiología , Adulto , Distribución por Edad , Anciano , Emigración e Inmigración , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Sistema de Registros , España/epidemiología , Adulto Joven
8.
PLoS Pathog ; 8(11): e1003005, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-23133388

RESUMEN

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.


Asunto(s)
Amiloide/química , Antibacterianos/química , Bacterias/química , Proteína Catiónica del Eosinófilo/química , Inmunidad Innata , Viabilidad Microbiana , Sustitución de Aminoácidos , Amiloide/genética , Amiloide/inmunología , Antibacterianos/inmunología , Bacterias/genética , Bacterias/inmunología , Infecciones Bacterianas , Proteína Catiónica del Eosinófilo/genética , Proteína Catiónica del Eosinófilo/inmunología , Humanos , Mutación Missense
9.
Lipids Health Dis ; 13: 31, 2014 Feb 11.
Artículo en Inglés | MEDLINE | ID: mdl-24512213

RESUMEN

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.


Asunto(s)
Suplementos Dietéticos , Ácidos Docosahexaenoicos/administración & dosificación , Ácido Eicosapentaenoico/administración & dosificación , Síndrome Metabólico/tratamiento farmacológico , Obesidad/tratamiento farmacológico , Grasa Abdominal/efectos de los fármacos , Grasa Abdominal/patología , Animales , Aorta/efectos de los fármacos , Aorta/patología , Ácidos Docosahexaenoicos/farmacología , Evaluación Preclínica de Medicamentos , Ácido Eicosapentaenoico/farmacología , Riñón/efectos de los fármacos , Riñón/patología , Hígado/efectos de los fármacos , Hígado/patología , Síndrome Metabólico/patología , Obesidad/patología , Ratas , Ratas Wistar
10.
Biochem J ; 456(1): 99-108, 2013 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-23962023

RESUMEN

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.


Asunto(s)
Péptidos Catiónicos Antimicrobianos/farmacología , Bacterias/efectos de los fármacos , Ribonucleasas/química , Aglutinación , Secuencia de Aminoácidos , Animales , Péptidos Catiónicos Antimicrobianos/síntesis química , Péptidos Catiónicos Antimicrobianos/química , Bacterias/inmunología , Secuencia Conservada , Evolución Molecular , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/efectos de los fármacos , Bacterias Grampositivas/inmunología , Hemólisis , Interacciones Huésped-Patógeno , Humanos , Inmunidad Innata , Liposomas/química , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Fosfatidilcolinas/química , Fosfatidilgliceroles/química , Filogenia , Ribonucleasas/inmunología , Ribonucleasas/fisiología , Ovinos
11.
Nutrients ; 16(8)2024 Apr 12.
Artículo en Inglés | MEDLINE | ID: mdl-38674832

RESUMEN

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.


Asunto(s)
Amina Oxidasa (conteniendo Cobre) , Histamina , Polimorfismo de Nucleótido Simple , Humanos , Amina Oxidasa (conteniendo Cobre)/genética , Amina Oxidasa (conteniendo Cobre)/sangre , Proyectos Piloto , Masculino , Femenino , Persona de Mediana Edad , Adulto , Histamina/sangre , Estudios Retrospectivos , Alelos , Prevalencia , Anciano , Predisposición Genética a la Enfermedad , Genotipo , Población Blanca/genética
12.
Antimicrob Agents Chemother ; 57(8): 3797-805, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23716047

RESUMEN

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.


Asunto(s)
Antibacterianos/farmacología , Proteína Catiónica del Eosinófilo/farmacología , Mycobacterium/efectos de los fármacos , Ribonucleasas/farmacología , Secuencia de Aminoácidos , Antibacterianos/síntesis química , Membrana Celular/efectos de los fármacos , Permeabilidad de la Membrana Celular , Activación Enzimática , Proteína Catiónica del Eosinófilo/síntesis química , Genes Sintéticos , Humanos , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana/efectos de los fármacos , Datos de Secuencia Molecular , Proteínas Recombinantes/farmacología , Ribonucleasas/síntesis química
13.
Bioinformatics ; 28(1): 130-1, 2012 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-22053077

RESUMEN

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.


Asunto(s)
Algoritmos , Antiinfecciosos/química , Péptidos/química , Péptidos Catiónicos Antimicrobianos/química , Internet , Lenguajes de Programación , Estructura Terciaria de Proteína , Programas Informáticos
14.
Lipids Health Dis ; 12: 140, 2013 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-24083393

RESUMEN

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.


Asunto(s)
Enfermedades Cardiovasculares/prevención & control , Ácidos Docosahexaenoicos/administración & dosificación , Ácido Eicosapentaenoico/administración & dosificación , Eritrocitos/efectos de los fármacos , Hígado/efectos de los fármacos , Animales , Apolipoproteínas/sangre , Proteína C-Reactiva/metabolismo , Catalasa/metabolismo , HDL-Colesterol/sangre , LDL-Colesterol/sangre , Dieta , Eritrocitos/metabolismo , Femenino , Glutatión Peroxidasa/metabolismo , Glutatión Reductasa/metabolismo , Hemoglobina Glucada/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Lipoproteínas LDL/sangre , Hígado/metabolismo , Malondialdehído/metabolismo , Ratas , Ratas Wistar , Superóxido Dismutasa/metabolismo , Triglicéridos/sangre
15.
Nutrients ; 15(12)2023 Jun 19.
Artículo en Inglés | MEDLINE | ID: mdl-37375711

RESUMEN

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.


Asunto(s)
Productos de la Carne , Animales , Humanos , Carne , Valor Nutritivo , Dieta Vegana , Vegetarianos
16.
J Struct Biol ; 179(1): 1-9, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22579681

RESUMEN

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.


Asunto(s)
Proteína Catiónica del Eosinófilo/química , Sulfatos/química , Secuencia de Aminoácidos , Dominio Catalítico , Cristalización , Proteína Catiónica del Eosinófilo/metabolismo , Glicosaminoglicanos/química , Glicosaminoglicanos/metabolismo , Humanos , Ligandos , Simulación de Dinámica Molecular , Datos de Secuencia Molecular , Unión Proteica , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo
17.
Antimicrob Agents Chemother ; 56(5): 2378-85, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22330910

RESUMEN

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.


Asunto(s)
Pared Celular/química , Proteína Catiónica del Eosinófilo/farmacología , Escherichia coli/efectos de los fármacos , Lipopolisacáridos/química , Oligopéptidos/farmacología , Aglutinación/efectos de los fármacos , Adhesión Bacteriana/efectos de los fármacos , Sitios de Unión , Conformación de Carbohidratos , Proteína Catiónica del Eosinófilo/química , Proteína Catiónica del Eosinófilo/metabolismo , Eosinófilos/citología , Eosinófilos/inmunología , Eosinófilos/metabolismo , Escherichia coli/genética , Escherichia coli/fisiología , Humanos , Lipopolisacáridos/metabolismo , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana/efectos de los fármacos , Modelos Moleculares , Oligopéptidos/química , Oligopéptidos/metabolismo , Unión Proteica , Estructura Terciaria de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , Relación Estructura-Actividad
18.
Biol Chem ; 393(8): 801-15, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22944682

RESUMEN

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.


Asunto(s)
Antibacterianos/química , Proteína Catiónica del Eosinófilo/química , Factores Inmunológicos/química , Secuencia de Aminoácidos , Animales , Antibacterianos/inmunología , Fenómenos Fisiológicos Bacterianos , Proteína Catiónica del Eosinófilo/inmunología , Interacciones Huésped-Patógeno , Humanos , Inmunidad Innata , Factores Inmunológicos/inmunología , Modelos Moleculares , Datos de Secuencia Molecular , Alineación de Secuencia
19.
Nutrients ; 14(9)2022 Apr 23.
Artículo en Inglés | MEDLINE | ID: mdl-35565742

RESUMEN

An underlying cause of histamine intolerance is diamine oxidase (DAO) deficiency, which leads to defective homeostasis and a higher systemic absorption of histamine. Impaired DAO activity may have a genetic, pharmacological or pathological origin. A recent proposal also suggests it can arise from an alteration in the gut microbiota, although only one study has explored this hypothesis to date. A greater abundance of histamine-secreting bacteria in the gut could lead to the development of histamine intolerance. Thus, the aim of this study was to characterize the composition of the intestinal microbiota of patients with histamine intolerance symptoms and compare it with that of healthy individuals. The study was performed by sequencing bacterial 16S rRNA genes (V3-V4 region) and analyzing the data using the EzBioCloud Database. Dysbiosis of the gut microbiota was observed in the histamine intolerance group who, in comparison with the healthy individuals, had a significantly lower proportion of Prevotellaceae, Ruminococcus, Faecalibacterium and Faecablibacterium prausnitzii, which are bacteria related to gut health. They also had a significantly higher abundance of histamine-secreting bacteria, including the genera Staphylococcus and Proteus, several unidentified genera belonging to the family Enterobacteriaceae and the species Clostridium perfringens and Enterococcus faecalis. A greater abundance of histaminogenic bacteria would favor the accumulation of high levels of histamine in the gut, its subsequent absorption in plasma and the appearance of adverse effects, even in individuals without DAO deficiency.


Asunto(s)
Amina Oxidasa (conteniendo Cobre) , Microbioma Gastrointestinal , Bacterias , Disbiosis/microbiología , Faecalibacterium , Heces/microbiología , Microbioma Gastrointestinal/genética , Histamina/efectos adversos , Humanos , ARN Ribosómico 16S/genética
20.
Front Nutr ; 9: 1018463, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36337620

RESUMEN

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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