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1.
Molecules ; 26(4)2021 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-33668684

RESUMO

The disposal of food waste is a current and pressing issue, urging novel solutions to implement sustainable waste management practices. Fish leftovers and their processing byproducts represent a significant portion of the original fish, and their disposal has a high environmental and economic impact. The utilization of waste as raw materials for the production of different classes of biofuels and high-value chemicals, a concept known as "biorefinery", is gaining interest in a vision of circular economy and zero waste policies. In this context, an interesting route of valorization is the extraction of omega-3 fatty acids (ω-3 FAs) for nutraceutical application. These fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have received attention over the last decades due to their beneficial effects on human health. Their sustainable production is a key process for matching the increased market demand while reducing the pressure on marine ecosystems and lowering the impact of waste production. The high resale value of the products makes this waste a powerful tool that simultaneously protects the environment and benefits the global economy. This review aims to provide a complete overview of the sustainable exploitation of fish waste to recover ω-3 FAs for food supplement applications, covering composition, storage, and processing of the raw material.


Assuntos
Suplementos Nutricionais , Ácidos Graxos Ômega-3/farmacologia , Peixes/metabolismo , Alimentos , Desenvolvimento Sustentável , Resíduos/análise , Animais , Ácidos Graxos Ômega-3/química , Óleos de Peixe/química , Óleos de Peixe/isolamento & purificação , Óleos de Peixe/farmacologia
2.
PLoS One ; 15(4): e0231494, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32298317

RESUMO

There is an increasing interest from the aquafeed industry in functional feeds containing selected additives that improve fish growth performance and health status. Functional feed additives include probiotics, prebiotics, organic acids, and phytogenics (substances derived from plants and their extracts). This study evaluated the effects of dietary inclusion of a mucilage extract rich in galactomannan oligosaccharides (GMOS), a mixture of garlic and labiatae-plants oils (PHYTO), and a combination of them (GMOSPHYTO), on gut microbiota composition of European sea bass (Dicentrarchus labrax) fed with a low fishmeal (FM) and fish oil (FO) diet. Three experimental diets and a control diet (plant-based formulation with 10% FM and 6% FO) were tested in a 63-days feeding trial. To analyze the microbiota associated to feeds and the intestinal autochthonous (mucosa-adhered) and allochthonous (transient) microbial communities, the Illumina MiSeq platform for sequencing of 16S rRNA gene and QIIME2 pipeline were used. Metabarcoding analysis of feed-associated bacteria showed that the microbial communities of control (CTRL) feed deeply differed from those of experimental diets. The number of reads was significantly lower in CTRL feed than in other feeds. The OTU (operational taxonomic unit) number was instead similar between the feeds, ranging from 42 to 50 OTUs. The variation of resident gut microbiota induced by diet was lower than the variation of transient intestinal microbiota, because feedstuffs are a major source of allochthonous bacteria, which can temporarily integrate into the gut transient microbiome. However, the composition of transient bacterial communities was not simply a mirror of feed-borne bacteria. Indeed, the microbial profile of feeds was different from both faecal and mucosa profiles. Our findings suggest that the dietary inclusion of GMOS (0.5%) and PHYTO (0.02%) in a low FM and FO diet induces changes in gut microbiota composition of European sea bass. However, if on allochthonous microbiota the combined inclusion of GMOS and PHYTO showed an antagonistic effect on bactericidal activity against Vibrionales, at mucosa level, only GMOSPHYTO diet increased the relative abundance of Bacteroidales, Lactobacillales, and Clostridiales resident bacterial orders. The main beneficial effects of GMOS and PHYTO on gut microbiota are the reduction of coliforms and Vibrionales bacteria, which include several potentially pathogenic species for fish, and the enrichment of gut microbiota composition with butyrate producer taxa. Therefore, these functional ingredients have a great potential to be used as health-promoting agents in the farming of European sea bass and other marine fish.


Assuntos
Bass/metabolismo , Dieta/veterinária , Suplementos Nutricionais , Óleos de Peixe/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacos , Mananas/farmacologia , Extratos Vegetais/farmacologia , Compostos Alílicos/farmacologia , Ração Animal , Animais , Aquicultura/métodos , Bass/crescimento & desenvolvimento , Bass/microbiologia , Galactose/análogos & derivados , Microbioma Gastrointestinal/genética , Óleos de Plantas/farmacologia , RNA Ribossômico 16S/genética , Sulfetos/farmacologia
3.
Mol Nutr Food Res ; 63(7): e1800813, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30632684

RESUMO

SCOPE: The mechanisms and involvement of uncoupling protein 1 (UCP1) in the protection from obesity and insulin resistance induced by intake of a high-fat diet rich in omega-3 (n-3) fatty acids are investigated. METHODS AND RESULTS: C57BL/6J mice are fed either a low-fat (control group) or one of two isocaloric high-fat diets containing either lard (HFD) or fish oil (HFN3) as fat source and evaluated for body weight, adiposity, energy expenditure, glucose homeostasis, and inguinal white and interscapular brown adipose tissue (iWAT and iBAT, respectively) gene expression, lipidome, and mitochondrial bioenergetics. HFN3 intake protected from obesity, glucose and insulin intolerances, and hyperinsulinemia. This is associated with increased energy expenditure, iWAT UCP1 expression, and incorporation of n-3 eicosapentaenoic and docosahexaenoic fatty acids in iWAT and iBAT triacylglycerol. Importantly, HFN3 is equally effective in reducing body weight gain, adiposity, and glucose intolerance and increasing energy expenditure in wild-type and UCP1-deficient mice without recruiting other thermogenic processes in iWAT and iBAT, such as mitochondrial uncoupling and SERCA-mediated calcium and creatine-driven substrate cyclings. CONCLUSION: Intake of a high-fat diet rich in omega-3 fatty acids protects both wild-type and UCP1-deficient mice from obesity and insulin resistance by increasing energy expenditure through unknown mechanisms.


Assuntos
Metabolismo Energético/efeitos dos fármacos , Óleos de Peixe/farmacologia , Intolerância à Glucose/dietoterapia , Obesidade/prevenção & controle , Proteína Desacopladora 1/genética , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Tecido Adiposo Branco/efeitos dos fármacos , Tecido Adiposo Branco/metabolismo , Animais , Dieta Hiperlipídica/efeitos adversos , Metabolismo Energético/genética , Ácidos Graxos Ômega-3/análise , Ácidos Graxos Ômega-3/farmacologia , Óleos de Peixe/química , Intolerância à Glucose/genética , Camundongos Endogâmicos C57BL , Camundongos Knockout , Obesidade/etiologia , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/genética , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Termogênese/efeitos dos fármacos , Termogênese/genética , Proteína Desacopladora 1/metabolismo
4.
Reumatismo ; 60(3): 174-9, 2008.
Artigo em Italiano | MEDLINE | ID: mdl-18854877

RESUMO

The beneficial properties of fish oil are well known and are related to its fatty acid composition rich in omega-3 polyunsaturated fatty acids. In the last years a variety of epidemiological and clinical studies have demonstrated the efficacy of fish oil supplementation in the rheumatic diseases, in particular in rheumatoid arthritis. The anti-inflammatory effects of fish oil are linked to the production of alternative eicosanoids, to the reduction of proinflammatory cytokines, to the inhibition of the activation of T lymphocytes and of catabolic enzymes. Fish oil supplementation could represent a valuable support to the traditional pharmacological treatment of rheumatoid arthritis.


Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Suplementos Nutricionais , Óleos de Peixe/uso terapêutico , Anti-Inflamatórios não Esteroides/economia , Anti-Inflamatórios não Esteroides/farmacocinética , Anti-Inflamatórios não Esteroides/farmacologia , Ensaios Clínicos como Assunto , Ciclosporina/farmacocinética , Custos de Medicamentos , Interações Medicamentosas , Ácido Eicosapentaenoico/farmacologia , Ácido Eicosapentaenoico/uso terapêutico , Ácidos Graxos Insaturados/farmacologia , Ácidos Graxos Insaturados/uso terapêutico , Óleos de Peixe/economia , Óleos de Peixe/farmacocinética , Óleos de Peixe/farmacologia , Humanos , Metotrexato/farmacocinética , Sulfassalazina/farmacocinética
5.
Am J Clin Nutr ; 88(3): 706-13, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18779287

RESUMO

BACKGROUND: It is suggested that a low intake of fish and/or n-3 PUFA is associated with depressed mood. However, results from epidemiologic studies are mixed, and randomized trials have mainly been performed in depressed patients, yielding conflicting results. OBJECTIVE: We investigated the effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on mental well-being in a double-blind, placebo-controlled trial. DESIGN: Independently living individuals (n = 302) aged > or =65 y were randomly assigned to consume 1800 mg/d EPA+DHA, 400 mg/d EPA+DHA, or placebo capsules for 26 wk. Changes in mental well-being were assessed as the primary outcome with the Center for Epidemiologic Studies Depression Scale (CES-D), Montgomery-Asberg Rating Scale (MADRS), Geriatric Depression Scale (GDS-15), and Hospital Anxiety and Depression Scale (HADS-A). RESULTS: Plasma concentrations of EPA+DHA increased by 238% in the high-dose and 51% in the low-dose fish-oil group compared with the placebo group, reflecting excellent compliance. Baseline CES-D scores ranged from 5.9 to 6.8 in the 3 groups and were not significantly different between groups. Mean changes in CES-D scores after 26 wk were -0.2, 0.2, and -0.4 (P = 0.87) in the high-dose fish oil, low-dose fish oil, and placebo groups, respectively. Treatment with neither 1800 mg nor 400 mg EPA+DHA differentially affected any of the measures of mental well-being after 13 or 26 wk of intervention compared with placebo. CONCLUSIONS: In this randomized, double-blind, placebo-controlled trial we observed no effect of EPA+DHA supplementation for 26 wk on mental well-being in the general older population studied. This trial was registered at clinicaltrials.gov as NCT00124852.


Assuntos
Óleos de Peixe/farmacologia , Saúde Mental , Entrevista Psiquiátrica Padronizada , Escalas de Graduação Psiquiátrica , Idoso , Ansiedade , Depressão , Suplementos Nutricionais , Ácidos Docosa-Hexaenoicos/administração & dosagem , Ácidos Docosa-Hexaenoicos/sangue , Ácidos Docosa-Hexaenoicos/farmacologia , Método Duplo-Cego , Ácido Eicosapentaenoico/administração & dosagem , Ácido Eicosapentaenoico/sangue , Ácido Eicosapentaenoico/farmacologia , Ácidos Graxos Ômega-3/administração & dosagem , Ácidos Graxos Ômega-3/sangue , Feminino , Óleos de Peixe/administração & dosagem , Humanos , Masculino , Placebos , Fatores Socioeconômicos
6.
Mol Carcinog ; 45(10): 741-51, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16683249

RESUMO

Our prior studies have shown that single topical treatment of repeated fish fried oil extract (RFFE), containing various polycyclic aromatic hydrocarbons (PAHs), to the dorsal epidermis of mice caused enhancement of DNA damage along with higher expression of p53 and p21WAF1 proteins and cell-cycle arrest. In the present study carcinogenic potential of repeated fish fried oil (RFFO) and RFFE was assessed. Single topical application of RFFO (100 microL/animal) and RFFE (100-500 microg/animal) to Swiss albino female mice resulted in significant induction (1.8- to 7.4-fold) of ornithine decarboxylase activity. Twice weekly topical application of methylcholanthrene (MCA) for 24 wk or single topical application of 7,12-dimethylbenzanthracene (DMBA) or RFFO or RFFE, as initiator followed by twice weekly application of 12-O-tetradecanoyl phorbol myristate acetate (TPA) as promoter for 24 wk, resulted in development of skin papillomas after 6, 7, 18, and 9 wk, respectively. The cumulative number of tumors in MCA, DMBA/TPA, RFFE (200 microg)/TPA, and RFFE (500 microg)/TPA groups were 276, 168, 34, and 58 after 24 wk while negligible or minimal initiating activity was noticed in RFFO/TPA group. No tumors were found in animals either given twice weekly topical application of RFFO or a single initiating dose of DMBA followed by twice weekly application of RFFO. Histopathology of skin of animals treated with RFFE/TPA showed marked proliferation of epidermal layers along with abnormal mitosis and multinucleated tumor appearance. Skin of animals in groups RFFO/TPA and DMBA/RFFO showed sloughing and regeneration of epidermal layers, oedema along with proliferation of fibroblasts. Histochemical localization of gamma-glutamyl transpeptidase was found to be substantially higher in skin of mice treated with RFFO/TPA and RFFE/TPA. Animals treated with RFFO/TPA, DMBA/RFFO, and RFFE/TPA resulted in significant induction of cutaneous aryl hydrocarbon hydroxylase (AHH) (421-432%), ethoxyresorufin-O-deethylase (252-316%), and glutathione S-transferase (133-245%) activities. Animals treated with RFFO/TPA, DMBA/RFFO, and RFFE/TPA led to significant reduction in glutathione content (39-44%) with a concomitant increase in lipid peroxidation (254-492%). Animals treated with RFFO/TPA and RFFE/TPA led a significant decrease in catalase (43-69%) and superoxide dismutase (20-31%) activities while glutathione reductase activity was found to be diminished (23-51%) in RFFO, RFFO/TPA, DMBA/RFFO, and RFFE/TPA treated groups. These results suggest that RFFE possess skin tumor initiating activity and that it may have weak promoting activity as well, which may involve free radicals.


Assuntos
Carcinógenos/toxicidade , Óleos de Peixe/toxicidade , Neoplasias Cutâneas/induzido quimicamente , Neoplasias Cutâneas/patologia , Administração Tópica , Albinismo , Animais , Biomarcadores , Carcinógenos/administração & dosagem , Carcinógenos/farmacologia , Indução Enzimática/efeitos dos fármacos , Feminino , Óleos de Peixe/administração & dosagem , Óleos de Peixe/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Metilcolantreno/farmacologia , Metilcolantreno/toxicidade , Camundongos , Ornitina Descarboxilase/metabolismo , Estresse Oxidativo , Neoplasias Cutâneas/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Fatores de Tempo
7.
Clin Diagn Lab Immunol ; 12(9): 1098-103, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16148177

RESUMO

Recent experimental observations have determined that long-chain n-3 polyunsaturated fatty acids suppress immune functions and are involved in the reduction of infectious disease resistance. BALB/c mice were fed for 4 weeks with one of four diets containing either olive oil (OO), fish oil (FO), hydrogenated coconut oil, or a low fat level. Interleukin-12p70 (IL-12p70), gamma interferon (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha) production in the sera of mice fed these diets and challenged with Listeria monocytogenes were determined by enzyme-linked immunosorbent assay. In addition, bacterial counts from spleens of mice were carried out at 24, 72, or 96 h of infection. Here, we quantified an initial diminution of production of both IL-12p70 and IFN-gamma, which appear to play an important role in the reduction of host resistance to L. monocytogenes infection. In addition, an efficient elimination of L. monocytogenes was observed in spleens of mice fed a diet containing OO at 96 h of infection, despite reductions in IL-12p70 and TNF-alpha production, suggesting an improvement of immune resistance. Overall, our results indicate that the initial reduction of both IL-12 and IFN-gamma production before L. monocytogenes infection represents the most relevant event that corroborates the impairment of immune resistance by n-3 polyunsaturated fatty acids during the different stages of infection. However, we speculate that the modulation of other cytokines must be also involved in this response, because the alteration of cytokine production in mice fed an FO diet in a late phase of L. monocytogenes infection was similar to that in mice fed OO, whereas the ability to eliminate this bacterium from the spleen was improved in the latter group.


Assuntos
Gorduras na Dieta/farmacologia , Interferon gama/sangue , Interleucina-12/sangue , Listeria monocytogenes/imunologia , Listeriose/imunologia , Subunidades Proteicas/sangue , Fator de Necrose Tumoral alfa/metabolismo , Animais , Biomarcadores/sangue , Óleo de Coco , Gorduras na Dieta/imunologia , Óleos de Peixe/farmacologia , Terapia de Imunossupressão/métodos , Interferon gama/metabolismo , Interleucina-12/metabolismo , Listeriose/sangue , Camundongos , Camundongos Endogâmicos BALB C , Azeite de Oliva , Óleos de Plantas/farmacologia , Subunidades Proteicas/metabolismo , Baço/imunologia , Baço/microbiologia
8.
Eur J Paediatr Neurol ; 5 Suppl A: 151-6, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11588988

RESUMO

The neuronal ceroid lipofuscinoses (NCLs) are inherited neurodegenerative diseases that occur in a number of animal species, including dogs. A study was conducted to determine whether the resupply of nutrients lost in NCL English Setter dogs would modify the course of the disease. Carnitine and polyunsaturated fatty acids have been reported to be reduced in NCL English Setters. Therefore, the normal laboratory diets of NCL dogs were supplemented with carnitine, fish oil and corn oil and the disease progression was compared with that of an untreated litter mate. The following specific prognostic indicators of NCL were monitored: cognitive function, brain atrophy, brain glucose metabolism and lifespan. Carnitine, with or without lipid supplements, dramatically delayed the progression of cognitive decline in NCL dogs. When fish oil and corn oil only were supplied, brain atrophy was reduced. A combination of all three supplements preserved cognitive function and increased lifespan by 10%. However, brain glucose hypometabolism and cerebral atrophy were not reduced. The results in this study indicated that the effectiveness of therapeutic interventions can be assessed by non-invasive methods at a relatively early stage of the disease process. Our study suggests that dietary supplementation with carnitine is a promising new approach for delaying or preventing the cognitive decline in dogs, and perhaps, with human NCL patients.


Assuntos
Carnitina/farmacologia , Óleo de Milho/farmacologia , Óleos de Peixe/farmacologia , Lipofuscinoses Ceroides Neuronais/dietoterapia , Animais , Ventrículos Cerebrais/patologia , Modelos Animais de Doenças , Cães , Glucose/metabolismo , Imageamento por Ressonância Magnética , Lipofuscinoses Ceroides Neuronais/patologia , Tomografia Computadorizada de Emissão
9.
Pol Merkur Lekarski ; 11(63): 233-8, 2001 Sep.
Artigo em Polonês | MEDLINE | ID: mdl-11761818

RESUMO

There is no specific and effective treatment for recurrent aphthous stomatitis (RAS), which is connected with its unexplained etiology. The purpose of this study was to determine the clinical efficiency and immunomodulation of the shark liver oil in treatment of recurrent aphthous stomatitis. Twenty-five patients with severe disease received treatment with shark liver oil during a three-month period. The frequency of occurrence of RAS decreased from 1.56 before treatment to 0.95 after treatment and the number of lesions per month was significantly reduced during the third month of treatment and two months after treatment. During two months after treatment 4 patients had no ulcers and an improvement was exhibited in all except 3 of the remaining patients. The treatment had an influence on some immunological parameters. A better response of neutrophils to OZ and PMA were seen. The B cell and T CD3/HLA DR+ cell percentage returned to normal values. A significantly increased percentage of T cells was observed as compared to the before treatment value. The level of C4 and the hemolytic activity of the complement system decreased after treatment and neared the normal values. The shark liver oil contains compounds that have a positive immunomodulation action and alleviate the course of disease.


Assuntos
Adjuvantes Imunológicos/uso terapêutico , Óleos de Peixe/uso terapêutico , Estomatite Aftosa/terapia , Adulto , Animais , Linfócitos B/metabolismo , Complexo CD3/metabolismo , Feminino , Óleos de Peixe/farmacologia , Antígenos HLA-DR/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Neutrófilos/metabolismo , Recidiva , Tubarões , Estomatite Aftosa/imunologia
10.
J Chromatogr B Biomed Sci Appl ; 748(1): 137-49, 2000 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-11092593

RESUMO

The effect of soybean oil (SO) and fish oil (FO) on the relative molecular species distribution of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylserine (PS) in Atlantic salmon head kidney was studied using normal-phase liquid chromatography coupled with negative ion electrospray tandem mass spectrometry. The conformation of identity of the phospholipid species was based on retention time, the mass of the [M-H]- ([M-15]- for PC) molecular ions and the carboxylate anion fragments in the product ion spectrum. The intensity ratio of sn-1/sn-2 fragment ions increased with increasing number of double bonds in the sn-2 acyl chain but was not affected by increasing number of double bonds in the sn-1 acyl chain of the species examined. The relative distribution of the molecular species was determined by multiple reaction monitoring of the carboxylate anion fragment from the sn-1 position. A total of 68 different phospholipid species were determined in the head kidney and the largest amount was found in PE (22 species). Depending on the diet, the main species identified in the different phospholipid classes were; PC 16:0/18:1, PE 16:0/22:6, PI 18:0/20:4 and PS 16:0/22:6. The SO diet significantly increased the 18:2, 20:3 and most 20:4 containing species and significantly reduced the 14:0 and most 20:5 and 22:6 fatty acid containing species. The increase of the 20:4 and the decrease of the 20:5 and 22:6 containing species were dependent on the fatty acid combination of the species.


Assuntos
Cromatografia Líquida/métodos , Óleos de Peixe/farmacologia , Rim/efeitos dos fármacos , Fosfolipídeos/metabolismo , Óleo de Soja/farmacologia , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Rim/química , Salmão
11.
Thromb Res ; 90(4): 163-9, 1998 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-9692615

RESUMO

The propensity to thrombosis in an individual or population represents a significant risk factor in coronary heart disease, that ultimately may result in acute myocardial infarction or unstable angina. A variety of currently available tests assess the relative potential for platelets to be activated and then aggregate, including agonist-dependent platelet aggregation or flow cytometric analysis of platelet activation. However, all of these methods have certain limitations, ranging from being poorly quantifiable with limited sensitivity, to the necessity for specialized equipment. In the present study, we describe the development of a simple whole blood, radiolabel assay that measures the surface expression of the alpha-granule protein, P-selectin, by activated platelets. This assay is performed in the presence of GP IIb-IIIa blockade to allow quantitation without interference by platelet aggregate formation, and thus directly measures agonist dose-response without complications arising from secondary activation mediated by GP IIb-IIIa. The sensitivity of this assay method to dietary manipulation was investigated by administration of fish oil capsules at a dose known to decrease platelet aggregation.


Assuntos
Plaquetas/fisiologia , Testes de Função Plaquetária/métodos , Adolescente , Adulto , Animais , Anticorpos Monoclonais/análise , Anticorpos Monoclonais/química , Plaquetas/ultraestrutura , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Colágeno/farmacologia , Gorduras na Dieta/farmacologia , Gorduras na Dieta/uso terapêutico , Relação Dose-Resposta Imunológica , Óleos de Peixe/farmacologia , Óleos de Peixe/uso terapêutico , Humanos , Radioisótopos do Iodo , Masculino , Camundongos , Selectina-P/biossíntese , Selectina-P/efeitos dos fármacos , Selectina-P/imunologia , Peptídeos/farmacologia , Ativação Plaquetária/efeitos dos fármacos , Agregação Plaquetária/efeitos dos fármacos , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/antagonistas & inibidores , Receptores de Trombina/agonistas , Receptores de Trombina/química , Reprodutibilidade dos Testes , Trombose/terapia
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