RESUMO
BACKGROUND: Eosinophilic asthma and nasal polyposis are hallmarks of aspirin-exacerbated respiratory disease (AERD), and IL-5 inhibition has been shown to provide therapeutic benefit. However, IL-5Rα is expressed on many cells in addition to eosinophils, and the mechanisms by which IL-5 inhibition leads to clinical benefit in eosinophilic asthma and nasal polyposis are unlikely to be due exclusively to antieosinophil effects. OBJECTIVE: We sought to identify the mechanisms by which anti-IL-5 treatment with mepolizumab improves respiratory inflammation in AERD. METHODS: The clinical characteristics, circulating granulocytes, nasal scraping transcripts, eosinophilic cationic protein, tryptase, and antibody levels, and urinary and nasal eicosanoid levels were measured for 18 subjects with AERD who were taking mepolizumab and compared with those of 18 matched subjects with AERD who were not taking mepolizumab. RESULTS: Subjects taking mepolizumab had significantly fewer peripheral blood eosinophils and basophils, and those cells that remained had higher surface CRTH2 expression than did the cells from subjects not taking mepolizumab. Nasal prostaglandin F2α, prostaglandin D2 metabolites, leukotriene B4, and thromboxane levels were lower in subjects taking mepolizumab, as were urinary levels of tetranor-prostaglandin D2 and leukotriene E4. The nasal epithelial cell transcripts that were overexpressed among subjects with AERD who were taking mepolizumab were enriched for genes involved in tight junction formation and cilium organization. Nasal and urinary prostaglandin E2, tryptase, and antibody levels were not different between the 2 groups. CONCLUSION: IL-5 inhibition in AERD decreases production of inflammatory eicosanoids and upregulates tight junction-associated nasal epithelial cell transcripts, likely due to decreased IL-5 signaling on tissue mast cells, eosinophils, and epithelial cells. These direct effects on multiple relevant immune cells contribute to the mechanism of benefit afforded by mepolizumab.
Assuntos
Anticorpos Monoclonais Humanizados/administração & dosagem , Asma Induzida por Aspirina , Basófilos , Eosinófilos , Pólipos Nasais , Adolescente , Adulto , Idoso , Asma Induzida por Aspirina/tratamento farmacológico , Asma Induzida por Aspirina/imunologia , Asma Induzida por Aspirina/urina , Basófilos/imunologia , Basófilos/patologia , Eicosanoides/imunologia , Eicosanoides/urina , Eosinófilos/imunologia , Eosinófilos/patologia , Feminino , Humanos , Interleucina-5/imunologia , Subunidade alfa de Receptor de Interleucina-5/imunologia , Masculino , Pessoa de Meia-Idade , Pólipos Nasais/tratamento farmacológico , Pólipos Nasais/imunologia , Pólipos Nasais/urinaRESUMO
Researchers have long assumed that systematic estrogen fading might contribute to the sustained progression of menopausal degenerate syndromes, although definitive evidence has not been presented. Whether such findings represent a causal contribution or are the result of opportunistic messengers sent from the reproductive system to the brain is also a vital question. We constructed a multiscale network of the ovariectomy (OVX) induced estrogen receptors depletion (ER-depletion) model and integrated targeted proteomic, targeted lipidomic, cytochemical, and histopathological data across three tissues from the ovariectomy rodent model. We found that compared to control rats, OVX rats showed increased renal and uterine prostaglandin D2 synthase (Ptgds) expression and decreased hypothalamic Ptgds expression, abnormal Ptgds metabolites, the degenerate renal function profiles and decreased cognitive ability (learning and memory) in Morris water maze test. Importantly, we observed a regulatory relationship among ER (particularly ERß), the degree of the pathological phenotype, learning behavior test and the 'hypothalamus-uterus-kidney (HUK) axis functions. Collectively, this study elucidates that ER depletion promoted HUK aging is mostly attributed to a renal ERß/Ptgds signalling imbalance.
Assuntos
Oxirredutases Intramoleculares/metabolismo , Rim/metabolismo , Metabolismo dos Lipídeos/genética , Lipocalinas/metabolismo , Menopausa/metabolismo , Receptores de Estrogênio/deficiência , Animais , Eicosanoides/sangue , Eicosanoides/urina , Feminino , Hipotálamo/enzimologia , Aprendizagem em Labirinto , Menopausa/genética , Ovariectomia , Proteoma , Ratos Sprague-Dawley , Transdução de Sinais , Útero/enzimologiaRESUMO
The urinary metabolites (Z)-7-[1R,2R,3R,5S)-3,5-dihydroxy-2-[(E,3S)-3-hydroxyoct-1-enyl]cyclopentyl]hept-5-enoic acid (8-iso-PGF2α), an F2-isoprostane and biomarker of oxidative damage, and "prostaglandin E2 metabolite" (PGE-M), a biomarker of inflammation, are elevated in cigarette smokers. However, there is little information in the literature on the longitudinal stability of these widely used biomarkers. In a large clinical trial involving 10 institutional sites, smokers were given, free of charge over a period of 20 weeks, Spectrum NRC600/601 research cigarettes containing 15.5 mg nicotine/g tobacco. All participants were instructed to smoke these cigarettes for the duration of the study. At weeks 4, 8, 12, 16, and 20, first morning urine voids were collected and analyzed for 8-iso-PGF2α and PGE-M using validated liquid chromatography-electrospray ionization-tandem mass spectrometry methods. The mean level of 8-iso-PGF2α at Week 4 was 1.34 ± 1.08 (S.D.) pmol/mg creatinine (N = 226) while that of PGE-M was 73.7 ± 113 (S.D.) pmol/mg creatinine (N = 232). The corresponding levels at Week 20 were 1.35 ± 0.93 (S.D.) pmol/mg creatinine (N = 209) for 8-iso-PGF2α and 74.2 ± 142 (S.D.) pmol/mg creatinine (N = 210) for PGE-M. There was variation in these values in the intervening weeks. The intra-class correlation coefficients (ICC) were 0.51 (95% CI, 0.45, 0.57) and 0.36 (0.30, 0.43), for 8-iso-PGF2α and PGE-M, respectively, indicating fair longitudinal stability for 8-iso-PGF2α and poorer longitudinal stability for PGE-M in cigarette smokers. Males had higher ICC values than females for both 8-iso-PGF2α and PGE-M. These results indicate that, in addition to cigarette smoking, endogenous processes of oxidative damage and inflammation influence the levels of these biomarkers over time among current smokers.
Assuntos
Biomarcadores/urina , Fumar Cigarros/urina , Eicosanoides/urina , Inflamação/urina , Estresse Oxidativo , Índice de Massa Corporal , Dinoprosta/análogos & derivados , Dinoprosta/urina , F2-Isoprostanos/urina , Feminino , Humanos , Masculino , Metaboloma , Pessoa de Meia-IdadeRESUMO
The eicosanoids are a family of lipid mediators of pain and inflammation involved in multiple pathologies, including asthma, hypertension, cancer, atherosclerosis, and neurodegenerative diseases. These signaling mediators act locally, but are rapidly metabolized and transported to the systemic circulation as a mixture of primary and secondary metabolites. Accordingly, urine has become a useful readily accessible biofluid for monitoring the endogenous synthesis of these molecules. Herein, we present the validation of a rapid, repeatable, and precise method for the extraction and quantification of 32 eicosanoid urinary metabolites by LC-MS/MS. For 12 out of 17 deconjugated glucuronide eicosanoids, there was no improvement in recovered signal. These metabolites cover the major synthetic pathways, including prostaglandins, leukotrienes, and isoprostanes. The method linearity was >0.99 for all metabolites analyzed, the limit of detection ranged from 0.05-5 ng/ml, and the average extraction recoveries were >90%. All analytes were stable for at least three freeze/thaw cycles. The method was formatted for large-scale analysis of clinical cohorts, and the long-term repeatability was demonstrated over 15 months of acquisition, evidencing high precision (CV <15%, except for tetranorPGEM and 2,3-dinor-11ß-PGF2α, which were <30%). The presented method is suitable for focused mechanistic studies as well as large-scale clinical and epidemiological studies that require repeatable methods capable of producing data that can be concatenated across multiple cohorts.
Assuntos
Eicosanoides/urina , Metabolômica/métodos , Asma/urina , Cromatografia Líquida de Alta Pressão , Humanos , Inflamação/urina , Isoprostanos/urina , Prostaglandinas/urina , Extração em Fase Sólida , Espectrometria de Massas em Tandem , Tromboxanos/urinaRESUMO
BACKGROUND: Nasal polyps influence the burden of aspirin-exacerbated respiratory disease (AERD) by contributing to eicosanoid production. AERD is diagnosed through graded aspirin challenges. It is not known how sinus surgery affects aspirin challenge outcomes. OBJECTIVE: To investigate the effects of endoscopic sinus surgery (ESS) on aspirin-induced reaction severity and on the levels of eicosanoids associated with these reactions. METHODS: Twenty-eight patients with AERD were challenged with aspirin before and 3 to 4 weeks after ESS. Respiratory parameters and plasma and urine levels of eicosanoids were compared before and after challenges. RESULTS: Before ESS, AERD diagnosis was confirmed in all study patients by aspirin challenges that resulted in hypersensitivity reactions. After ESS, reactions to aspirin were less severe in all patients and 12 of 28 patients (43%, P < .001) had no detectable reaction. A lack of clinical reaction to aspirin was associated with lower peripheral blood eosinophilia (0.1 K/µL [interquartile range (IQR) 0.1-0.3] vs 0.4 K/µL [IQR 0.2-0.8]; P = .006), lower urinary leukotriene E4 levels after aspirin challenge (98 pg/mg creatinine [IQR 61-239] vs 459 pg/mg creatinine [IQR 141-1344]; P = .02), and lower plasma prostaglandin D2 to prostaglandin E2 ratio (0 [±0] vs 0.43 [±0.2]; P = .03), compared with those who reacted. CONCLUSIONS: Sinus surgery results in decreased aspirin sensitivity and a decrease in several plasma and urine eicosanoid levels in patients with AERD. Diagnostic aspirin challenges should be offered to patients with suspected AERD before ESS to increase diagnostic accuracy. Patients with established AERD could undergo aspirin desensitizations after ESS as the severity of their aspirin-induced hypersensitivity reactions lessens.
Assuntos
Asma Induzida por Aspirina , Endoscopia , Procedimentos Cirúrgicos Nasais , Adulto , Aspirina/efeitos adversos , Asma Induzida por Aspirina/sangue , Asma Induzida por Aspirina/metabolismo , Asma Induzida por Aspirina/fisiopatologia , Asma Induzida por Aspirina/urina , Eicosanoides/sangue , Eicosanoides/urina , Feminino , Volume Expiratório Forçado , Humanos , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/metabolismo , Seios Paranasais , Índice de Gravidade de DoençaRESUMO
BACKGROUND: Leukotriene (LT) E4 is the final active metabolite among the cysteinyl leukotrienes (CysLTs). Animal studies have identified a distinct LTE4 receptor, suggesting that current cysteinyl leukotriene type 1 (CysLT1) receptor antagonists can provide incomplete inhibition of CysLT responses. OBJECTIVE: We tested this hypothesis by assessing the influence of the CysLT1 antagonist montelukast on responses induced by means of inhalation of LTE4 in asthmatic patients. METHODS: Fourteen patients with mild intermittent asthma and 2 patients with aspirin-exacerbated respiratory disease received 20 mg of montelukast twice daily and placebo for 5 to 7 days in a randomized, double-blind, crossover study (NCT01841164). The PD20 value was determined at the end of each treatment period based on an increasing dose challenge. Measurements included lipid mediators in urine and sputum cells 4 hours after LTE4 challenge. RESULTS: Montelukast completely blocked LTE4-induced bronchoconstriction. Despite tolerating an at least 10 times higher dose of LTE4 after montelukast, there was no difference in the percentage of eosinophils in sputum. Urinary excretion of all major lipid mediators increased after LTE4 inhalation. Montelukast blocked release of the mast cell product prostaglandin (PG) D2, as well as release of PGF2α and thromboxane (Tx) A2, but not increased excretion of PGE2 and its metabolites or isoprostanes. CONCLUSION: LTE4 induces airflow obstruction and mast cell activation through the CysLT1 receptor.
Assuntos
Acetatos/uso terapêutico , Asma/tratamento farmacológico , Asma/fisiopatologia , Broncoconstritores/administração & dosagem , Eicosanoides/administração & dosagem , Antagonistas de Leucotrienos/uso terapêutico , Mastócitos/efeitos dos fármacos , Quinolinas/uso terapêutico , Receptores de Leucotrienos/fisiologia , Adulto , Aspirina/efeitos adversos , Asma/urina , Broncoconstrição/efeitos dos fármacos , Broncoconstritores/urina , Estudos Cross-Over , Ciclopropanos , Método Duplo-Cego , Eicosanoides/urina , Feminino , Humanos , Masculino , Mastócitos/fisiologia , Pessoa de Meia-Idade , Sulfetos , Adulto JovemRESUMO
A preceding untargeted metabolic fingerprinting approach in our lab followed by targeted fatty acid analysis revealed alterations in arachidonic acid metabolism in samples derived from a diet-controlled smoking cessation study in which compliant subjects ( N = 39) quit smoking at baseline and were followed over the course of 3 months. Consequently, urinary eicosanoids were evaluated by means of a validated LC-MS/MS method. A significant decrease was obtained for the prostaglandins PGF2α, 8-iso-PGF2α, thromboxane 2,3-d-TXB2, and leukotriene E4 upon quitting smoking. These findings indicate a partial recovery of smoking-induced alterations in the eicosanoid profile due to a reduction in oxidative stress and the inflammatory response.
Assuntos
Fumar Cigarros/urina , Eicosanoides/urina , Metaboloma , Abandono do Uso de Tabaco , Cromatografia Líquida/métodos , Fumar Cigarros/metabolismo , Eicosanoides/metabolismo , Humanos , Masculino , Metabolômica/métodos , Estresse Oxidativo , Espectrometria de Massas em Tandem/métodos , Abandono do Uso de Tabaco/métodosRESUMO
BACKGROUND: Treatment with acetylsalicylic acid (ASA) after desensitization may be a therapeutic option in patients with nonsteroidal anti-inflammatory drug exacerbated respiratory disease (NERD). The mechanisms that lead to improvement in rhinosinusitis and asthma symptoms remain unknown. AIM: To attribute the documented clinical effects of ASA treatment of chronic rhinosinusitis and/or asthma to the release of eicosanoid metabolites in urine. METHODS: Fourteen patients with NERD were successfully desensitized, and, eventually, eight patients were treated with 650 mg of ASA daily for 3 months. In addition to clinical assessments, nuclear magnetic resonance imaging and smell test were performed before and after treatment with ASA. Venous blood and urine were collected before desensitization and after 1 and 3 months of treatment. The levels of urinary leukotrienes (LT) (cysteinyl LT and LTE4) and tetranor PGDM (metabolite of prostaglandin D2) were measured by enzyme-linked immunosorbent assay. RESULTS: Treatment with ASA after desensitization alleviated symptoms of rhinosinusitis, improved nasal patency (mean, 50% decrease in peak nasal inspiratory flow) and sense of smell (fourfold increase in smell test score) in as early as 4 weeks. Clinical improvements were not accompanied by any change in sinonasal mucosa thickness as assessed with nuclear magnetic resonance. Urinary cysteinyl LTs, LTE4, and prostaglandin D2 metabolite remained relatively stable during ASA treatment and did not correlate with clinical improvements. Desensitization was associated with a progressive decrease of urinary creatinine. CONCLUSION: Clinical improvement in rhinosinusitis and/or asthma after ASA desensitization was not related to concentrations of urinary eicosanoid metabolites. A decrease of urinary creatinine requires further study to determine the renal safety of long-term treatment with ASA after desensitization.
Assuntos
Aspirina/uso terapêutico , Creatina/urina , Dessensibilização Imunológica/métodos , Hipersensibilidade a Drogas/terapia , Eicosanoides/urina , Adulto , Anti-Inflamatórios não Esteroides/efeitos adversos , Aspirina/imunologia , Aspirina/farmacologia , Asma/urina , Humanos , Leucotrienos/urina , Prostaglandina D2/análogos & derivados , Prostaglandina D2/urina , Doenças Respiratórias/induzido quimicamente , Doenças Respiratórias/imunologia , Doenças Respiratórias/urina , Sinusite/urinaRESUMO
BACKGROUND: Aspirin desensitization followed by high-dose aspirin therapy is routinely performed for patients with aspirin-exacerbated respiratory disease (AERD). Little is known about the contributions of mediators other than cysteinyl leukotrienes to aspirin reactions and to the therapeutic benefit of high-dose aspirin therapy. OBJECTIVE: We investigated differences in urinary eicosanoid metabolite levels and blood eosinophil counts in patients with AERD who tolerate and those who fail aspirin desensitization and also in patients with AERD who were successfully treated with high-dose aspirin therapy. METHODS: Twenty-nine patients with AERD were stratified into those who tolerated aspirin desensitization (group I) and those who did not (group II). Urine was analyzed for eicosanoid metabolites at baseline, during aspirin reactions, and during high-dose aspirin therapy. Blood was analyzed for cell differentials at baseline and during aspirin therapy. RESULTS: Basal prostaglandin D2 metabolite (PGD-M; 13.6 ± 2.7 vs 7.0 ± 0.8 pmol/mg creatinine [Cr], P < .05) and thromboxane metabolite (TX-M; 1.4 ± 0.3 vs 0.9 ± 0.1 pmol/mg Cr, P < .01) levels were higher in group II than in group I. During aspirin reactions, PGD-M levels remained unchanged, whereas TX-M levels (0.7 ± 0.1 pmol/mg Cr, P = .07) tended to decrease in group I. In contrast, PGD-M levels increased dramatically in group II (61.3 ± 19.9 pmol/mg Cr, P < .05), whereas TX-M levels did not change. The decrease in FEV1 inversely correlated with basal urinary levels of both leukotriene E4 and PGD-M. Blood eosinophil and basophil levels increased and urinary PGD-M levels (2.2 ± 0.8 pmol/mg Cr, P < .001) decreased on 2 months of high-dose aspirin therapy in group I. CONCLUSION: Failure to tolerate aspirin desensitization in a subset of patients with AERD is associated with prostaglandin D2 overproduction. The increase in blood eosinophil and basophil counts during high-dose aspirin therapy might reflect the functional consequences of decreased prostaglandin D2 release and the therapeutic benefit of aspirin.
Assuntos
Aspirina/efeitos adversos , Eicosanoides/imunologia , Doenças Respiratórias/induzido quimicamente , Doenças Respiratórias/imunologia , Adulto , Aspirina/imunologia , Basófilos , Dessensibilização Imunológica , Hipersensibilidade a Drogas/etiologia , Hipersensibilidade a Drogas/imunologia , Hipersensibilidade a Drogas/terapia , Eicosanoides/urina , Eosinófilos , Feminino , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Doenças Respiratórias/urinaRESUMO
Eicosanoids are key mediators and regulators of inflammation and oxidative stress often used as biomarkers for diseases and pathological conditions such as cardiovascular and pulmonary diseases and cancer. Analytically, comprehensive and robust quantification of different eicosanoid species in a multi-method approach is problematic because most of these compounds are relatively unstable and may differ in their chemical properties. Here we describe a novel ultra-performance liquid chromatography-selected reaction monitoring mass spectroscopy (UPLC-SRM/MS) method for simultaneous quantification of key urinary eicosanoids, including the prostaglandins (PG) tetranor PGE-M, 8-iso-, and 2,3-dinor-8-iso-PGF(2α); the thromboxanes (TXs) 11-dehydro- and 2,3-dinor-TXB2; leukotriene E4; and 12-hydroxyeicosatetraenoic acid. In contrast to previous methods, which used time-consuming and complex solid phase extraction, we prepared samples with a simple liquid/liquid extraction procedure. Because collision-induced dissociation produced characteristic product ions for all analytes, no derivatization step for SRM/MS analysis was necessary. Analytes were separated with a short UPLC reversed-phase column (1.7 µm particles), allowing shorter run times than conventional HPLC columns. The method was validated and applied to human urine samples showing excellent precision, accuracy, detection limits, and robustness. In summary, the developed method allows robust and sensitive profiling of urinary eicosanoid species, making it a useful and valuable tool for biomarker profiling in clinical/toxicological studies.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Eicosanoides/urina , Espectrometria de Massas em Tandem/métodos , Urinálise/métodos , Métodos Analíticos de Preparação de Amostras , Calibragem , Eicosanoides/química , Eicosanoides/isolamento & purificação , Humanos , Mediadores da Inflamação/química , Mediadores da Inflamação/isolamento & purificação , Mediadores da Inflamação/urina , Reprodutibilidade dos Testes , Fumar/urinaRESUMO
BACKGROUND: Cardiovascular disease (CVD) risk can be underestimated in HIV-infected patients receiving antiretroviral therapy (ART). Novel CVD risk markers in this population are needed. We hypothesized that eicosanoid metabolite production is increased with metabolic complications of ART. Our objective was to determine relationships between urine eicosanoids and traditional CVD risk factors in a cohort of HIV-infected persons receiving ART. METHODS: Cross-sectional analysis of 107 individuals from a prospective cohort study with urine eicosanoids (isoprostane [15-F(2t)-IsoP], prostaglandin-E metabolite [PGE-M], thromboxane metabolite [11dTxB(2)], prostacyclin metabolite [PGI-M]) determined by gas or liquid chromatography-mass spectrometry. RESULTS: 15-F(2t)-IsoP was higher (P=0.003), 11dTxB(2) tended to be higher (P=0.07) and PGE-M was lower (P=0.003) in females than in males. The overall median Framingham score was 4 (IQR 1-7). In multivariable analyses adjusting for age, CD4(+) T-cells, smoking status, non-steroidal anti-inflammatory drug use, aspirin use and body mass index (BMI), associations included: higher 15-F(2t)-IsoP with female sex (P=0.004) and current smoking (P=0.04), lower PGE-M with female sex (P=0.005) and higher BMI (P=0.03), higher 11dTxB(2) with increasing age (P=0.02) and current smoking (P=0.04), lower 11dTxB(2) with higher BMI (P=0.02), and higher PGI-M with current smoking (P=0.04). CONCLUSIONS: In this pilot study of predominantly virologically suppressed HIV-infected individuals on ART, there were sex-specific differences in urinary eicosanoids, with females having more risk-associated parameters despite a low Framingham score. Eicosanoids might be useful CVD biomarkers in ART-treated, HIV-infected patients. Future studies should examine eicosanoids while assessing effects of specific ART regimens and targeted interventions on CVD outcomes.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Biomarcadores/urina , Doenças Cardiovasculares/sangue , Eicosanoides/urina , Infecções por HIV/complicações , Inibidores da Transcriptase Reversa/uso terapêutico , Caracteres Sexuais , Adulto , Fármacos Anti-HIV/efeitos adversos , Fármacos Anti-HIV/farmacologia , Doenças Cardiovasculares/etiologia , Feminino , Infecções por HIV/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Estudos Prospectivos , Inibidores da Transcriptase Reversa/efeitos adversos , Inibidores da Transcriptase Reversa/farmacologia , Fatores de RiscoRESUMO
Substantial evidence relates increased sex hormone concentrations with increased breast cancer risk. Varying omega-3 (n-3) fatty acid (FA) intake may lead to alterations in eicosanoid balance and changes in circulating sex hormones that reduce risk. To clarify effects of dietary fat and n-3 FA intake on breast cancer risk markers, circulating sex hormones and urinary eicosanoids were measured in response to controlled feeding of diets designed to increase plasma concentrations of n-3 FA. A controlled cross-over feeding trial in postmenopausal women was conducted using 3 diets: high fat (HF; 40% energy from fat), low fat (LF; 20% energy from fat), and low fat plus n-3 FA (LFn3; 20% of energy from fat plus 3% of energy from n-3 FA) in 8-wk feeding periods. Plasma phospholipid fatty acid n-3 increased with the LFn3 relative to HF and LF (P < 0.0001). Plasma estradiol increased by 51% with HF (P = 0.03). Urinary prostaglandin E metabolite increased with HF relative to LF (P = 0.02) and urinary 11-dehydro-thromboxane B(2) increased with HF (P = 0.01). These results do not support a role of n-3 FA in the reduction of sex hormone levels.
Assuntos
Gorduras na Dieta/administração & dosagem , Eicosanoides/urina , Ácidos Graxos Ômega-3/administração & dosagem , Hormônios Esteroides Gonadais/sangue , Pós-Menopausa , Idoso , Peso Corporal , Neoplasias da Mama , Estudos Cross-Over , Dieta , Ingestão de Energia , Feminino , Humanos , Modelos Lineares , Pessoa de Meia-Idade , Fosfolipídeos/sangue , Fatores de Risco , Inquéritos e QuestionáriosRESUMO
BACKGROUND: It has recently demonstrated that a free radical-mediated pathway generates prostaglandins (PGs) and the corresponding prostaglandin enantiomers (ent-PGs). Aspirin-intolerant asthma and anaphylaxis accompany PGD(2) overproduction, possibly associated with mast cell activation via the COX pathway. However, free radical-mediated PG generation in the pathophysiology of these diseases, which can be demonstrated by measuring urinary ent-PGF(2)alpha, has not been reported. OBJECTIVES: To evaluate the characteristic profile of eicosanoid generation via the COX and/or free radical-mediated pathway underlying aspirin-intolerant asthma and anaphylaxis. METHODS: A comparative group analysis consisted of asthma (n = 17) and anaphylaxis (n = 8, none with aspirin-induced anaphylaxis) cases. Urinary eicosanoid concentrations were quantified as follows: 2,3-dinor-9alpha,11beta-PGF(2) by gas chromatography-mass spectrometry; leukotriene E(4), 9alpha,11beta-PGF(2), and PGs by enzyme immunoassay. RESULTS: 2,3-Dinor-9alpha,11beta-PGF(2) is a more predominant PGD(2) metabolite in urine than 9alpha,11beta-PGF(2). At baseline, the aspirin-intolerant asthma group (n = 10) had significantly higher leukotriene E(4) and lower PGE(2) concentrations in urine than the aspirin-tolerant asthma group. During the reaction, the urinary concentrations of leukotriene E(4) and PGD(2) metabolites correlatively increased, but with markedly different patterns of the mediator release, in the aspirin-intolerant asthma group and the anaphylaxis group, respectively. The urinary PGD(2) metabolites and primary PGs were significantly decreased in the aspirin-tolerant asthma group. Urinary ent-PGF(2)alpha concentrations were significantly increased in the anaphylaxis group but not the aspirin-intolerant asthma group. CONCLUSIONS: When assessed by urinary 2,3-dinor-9alpha,11beta-PGF(2), PGD(2) overproduction during aspirin-intolerant bronchoconstriction was clearly identified, regardless of COX inhibition. It is evident that free radical-mediated PG generation is involved in the pathophysiology of anaphylaxis.
Assuntos
Anafilaxia/fisiopatologia , Aspirina/efeitos adversos , Asma/fisiopatologia , Biomarcadores/urina , Cisteína/urina , Dinoprosta/urina , Leucotrienos/urina , Adulto , Idoso , Anafilaxia/imunologia , Asma/induzido quimicamente , Asma/imunologia , Tempo de Sangramento , Eicosanoides/urina , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Immunoaffinity chromatography (IAC), mass spectrometry and especially tandem mass spectrometry (MS/MS) represent the most efficient and reliable analytical techniques for specific isolation, unequivocal identification and accurate quantification of numerous natural and synthetic substances in biological samples. This review article focuses on the combined use of these outstanding methodologies in basic and clinical research and in life sciences for the quantitative analysis of low- and high-molecular mass biomarkers, drugs and toxins in urine, plasma or serum samples, in tissue and other biologicals systems published in the last decade. The analytes discussed in some detail include the biomarkers of oxidative stress 15(S)-8-iso-prostaglandin F(2alpha) {15(S)-8-iso-PGF(2alpha)} and 3-nitrotyrosine, the major urinary metabolite of the lipid mediators cysteinyl leukotrienes, i.e., the leukotriene E(4) (LTE(4)), melatonin, and the major collagen type II neoepitope peptide in human urine.
Assuntos
Biomarcadores/análise , Cromatografia de Afinidade/métodos , Técnicas de Imunoadsorção , Espectrometria de Massas/métodos , Preparações Farmacêuticas/análise , Toxinas Biológicas/análise , Animais , Eicosanoides/sangue , Eicosanoides/urina , Humanos , Preparações Farmacêuticas/sangue , Preparações Farmacêuticas/urina , Tirosina/análogos & derivados , Tirosina/sangueRESUMO
BACKGROUND: The precise pathway by which alcohol causes the characteristic features of fetal alcohol spectrum disorders is unknown. Proposed mechanisms for fetal injury from maternal alcohol use include cellular damage from oxidative stress and impaired fetal oxygenation related to maternal systemic vasoconstriction. Our objective was to compare the levels of urinary markers of oxidative stress and systemic vasoconstriction between women consuming large amounts of alcohol during pregnancy and women who did not drink alcohol during pregnancy. METHODS: Pregnant women consuming > or =48 g alcohol per day (n = 29) on average and pregnant women who abstained from alcohol use (n = 39) were identified using detailed interviews and home visits. Random maternal urine specimens were collected. Urinary levels of the oxidative stress marker, 8-isoprostane F2alpha, and of the vasoactive prostaglandin metabolites, 2,3-dinor-6-keto-prostaglandin F1alpha (a vasodilator) and 11-dehydro-thromboxane B2 (a vasoconstrictor), were measured using mass spectrometric methods. All analyte levels were corrected for urinary creatinine. RESULTS: In crude analyses, there was no significant difference in 8-isoprostane F2alpha between pregnant drinkers and nondrinkers (2.16 vs. 2.08 ng/mg creatinine, respectively, p = 0.87). There were no significant differences between the drinking and nondrinking groups in levels of 2,3-dinor-6-keto-prostaglandin F1alpha (1.03 vs. 1.17 ng/mg creatinine, respectively, p = 0.50), 11-dehydro-thromboxane B2 (0.72 vs. 0.59 ng/mg creatinine, respectively, p = 0.21), or the ratio of vasodilatory metabolite to vasoconstrictive metabolite (1.73 vs. 2.72, respectively, p = 0.14). Adjusting for maternal age, marital status, smoking, and gestational age at sampling did not substantially alter the results. CONCLUSION: Our results show no difference in levels of urinary eicosanoid markers of oxidative stress and systemic vasoconstriction between pregnant women who drink heavily and pregnant women who abstain. These findings speak against a role for maternal oxidative stress or systemic vasoconstriction in the pathogenesis of alcohol damage to the fetus.
Assuntos
Consumo de Bebidas Alcoólicas/fisiopatologia , Eicosanoides/urina , Transtornos do Espectro Alcoólico Fetal/etiologia , Isoprostanos/urina , Estresse Oxidativo/fisiologia , Vasoconstrição/fisiologia , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/urina , Adolescente , Adulto , Consumo de Bebidas Alcoólicas/efeitos adversos , Estudos de Casos e Controles , Depressores do Sistema Nervoso Central/efeitos adversos , Estudos de Coortes , Dinoprosta/análogos & derivados , Dinoprosta/urina , Etanol/efeitos adversos , Feminino , Transtornos do Espectro Alcoólico Fetal/fisiopatologia , Seguimentos , Humanos , Gravidez , Tromboxano B2/análogos & derivados , Tromboxano B2/urina , Adulto JovemRESUMO
Conjugated linoleic acid (CLA) refers to a group of positional and geometric isomers of linoleic acid and has been shown to suppress the development of atherosclerosis in experimental models. However, the mechanism involved is unclear although it is believed it may act as a cyclooxygenase inhibitor or as an agonist of the nuclear receptors, peroxisome proliferator activated receptors (PPARs). In this study, we examined the effect of cis-9,trans-11:trans-10,cis-12-CLA (80:20 blend) on the regression of pre-established atherosclerosis. ApoE(-/-) mice fed a 1% cholesterol diet were randomized at 8 weeks to continue receiving the diet supplemented with 1% control saturated fat or 1% CLA blend for a further 8 weeks. CLA supplementation did not simply prevent progression but induced almost complete resolution of atherosclerosis. Although CLA inhibited platelet deposition, as detected by staining of platelet glycoprotein alpha11b beta111a, it did not inhibit COX-mediated generation of prostaglandins in this model. However, PPARalpha and PPARgamma expression was increased in the aorta of the CLA-treated animals. This was coincident with decreased macrophage accumulation and decreased expression of the macrophage scavenger receptor CD36 and increased apoptosis in the aorta in vivo. CLA induces the resolution of atherosclerosis by negatively regulating the expression of pro-inflammatory genes and inducing apoptosis in the atherosclerotic lesion.
Assuntos
Aterosclerose/tratamento farmacológico , Ácidos Linoleicos Conjugados/farmacologia , Animais , Aorta/metabolismo , Apoptose , Aterosclerose/metabolismo , Plaquetas/metabolismo , Linhagem Celular , Colesterol/metabolismo , Eicosanoides/urina , Epoprostenol/metabolismo , Homozigoto , Humanos , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Monócitos/metabolismo , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Tromboxanos/metabolismo , Triglicerídeos/metabolismoRESUMO
BACKGROUND: Vasculitides are classified on the basis of the type of cell involved, namely, eosinophilic vasculitides such as Churg-Strauss syndrome (CSS) and noneosinophilic vasculitides. However, knowledge on inflammatory mediators and oxidative tissue damage associated with vasculitides is insufficient. OBJECTIVE: We measured the urinary concentrations of inflammatory mediators and tyrosine derivatives to assess biomarkers associated with the pathophysiology of vasculitides. METHODS: Urine was collected from 9 patients with CSS during acute exacerbation and during clinical remission, 24 patients with rheumatoid arthritis in stable condition, and 8 patients with vasculitis diseases (VDs) during acute exacerbation. Leukotriene E 4 (LTE 4 ), 9alpha,11beta prostaglandin F 2 , and eosinophil-derived neurotoxin (EDN) concentrations were determined by enzyme immunoassay. 3-Bromotyrosine (BrY) and 3-chlorotyrosine (ClY) concentrations were determined by gas chromatography-mass spectrometry. RESULTS: The urinary LTE 4 , EDN, BrY, and ClY concentrations were significantly higher in the patients with CSS during acute exacerbation than in healthy control subjects and, except for urinary ClY concentration, significantly decreased during clinical remission. The urinary EDN and BrY concentrations were significantly higher in patients with CSS during acute exacerbation than in patients with VD during acute exacerbation. Only urinary LTE 4 concentration was significantly different between the patients with rheumatoid arthritis in stable condition and the patients with VD during acute exacerbation. CONCLUSION: Oxidative tissue damage caused by eosinophil peroxidase is a pathophysiological characteristic of eosinophil-associated diseases such as CSS. Urinary LTE 4 concentration may reflect a pathophysiological event involved in eosinophilic and noneosinophilic vasculitides. Cysteinyl-leukotriene pathways are potential therapeutic targets for small-vessel vasculitides.
Assuntos
Eicosanoides/urina , Tirosina/análogos & derivados , Tirosina/urina , Vasculite/urina , Adulto , Idoso , Síndrome de Churg-Strauss/urina , Neurotoxina Derivada de Eosinófilo/urina , Feminino , Humanos , Leucotrieno E4/urina , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Although many studies have assumed that the overproduction of cysteinyl- leukotrienes (cys-LTs) and an imbalance of arachidonic acid metabolism may be plausible causes for the pathogenesis of aspirin-intolerant asthma (AIA), there has been little experimental evidence to substantiate this notion in lower airways of patients with AIA. OBJECTIVES: The purpose of this study was to compare the eicosanoid concentrations in sputum and urine from patients with AIA. METHODS: The concentrations of sputum cys-LTs, prostaglandin E2 (PGE2), PGF2alpha, PGD2 and thromboxane B2 were measured to assess local concentrations of eicosanoids in patients with AIA and in those with aspirin-tolerant asthma (ATA). The concentrations of two urinary metabolites, leukotriene E4 (LTE4) and 9alpha11betaPGF2, were also measured to corroborate the relationship between the eicosanoid biosynthesis in the whole body and that in lower airways. RESULTS: The concentration of PGD2 in sputum was significantly higher in patients with AIA than in those with ATA (median, 5.3 pg/mL vs. 3.1 pg/mL, P < 0.05), but there was no significant difference in the concentration of the corresponding metabolite, 9alpha11betaPGF2, between the two groups. No differences were noted in the concentrations of other prostanoids in sputum between the two groups. The sputum cys-LT concentrations showed no differences between the two groups, in spite of the observation that the concentration of urinary LTE4 was significantly higher in patients with AIA than in those with ATA (median, 195.2 pg/mg-cre vs. 122.1 pg/mg-cre, P < 0.05). There was a significant correlation among the concentration of cys-LTs, the number of eosinophils and the concentration of eosinophil-derived neurotoxin (EDN) in sputum. CONCLUSION: The urinary concentration of LTE4 does not necessary reflect cys-LT biosynthesis in lower airways. A significantly higher concentration of PGD2 in sputum from patients with AIA suggests the possible ongoing mast cell activation in lower airways.
Assuntos
Anti-Inflamatórios não Esteroides/efeitos adversos , Aspirina/efeitos adversos , Asma/metabolismo , Hipersensibilidade a Drogas/metabolismo , Eicosanoides/análise , Asma/imunologia , Estudos de Casos e Controles , Hipersensibilidade a Drogas/imunologia , Eicosanoides/urina , Neurotoxina Derivada de Eosinófilo , Eosinófilos , Feminino , Humanos , Contagem de Leucócitos , Leucotrieno E4/urina , Masculino , Pessoa de Meia-Idade , Prostaglandina D2/análise , Rinite/imunologia , Rinite/metabolismo , Ribonucleases/análise , Sinusite/imunologia , Sinusite/metabolismo , Escarro/químicaAssuntos
Eicosanoides/sangue , Transplante de Rim/fisiologia , Rim , Soluções para Preservação de Órgãos , Preservação de Órgãos/métodos , 6-Cetoprostaglandina F1 alfa/sangue , 6-Cetoprostaglandina F1 alfa/urina , Adenosina , Alopurinol , Análise de Variância , Biomarcadores/sangue , Cadáver , AMP Cíclico/sangue , AMP Cíclico/urina , Eicosanoides/urina , Glutationa , Humanos , Soluções Hipertônicas , Insulina , Leucotrieno E4/sangue , Período Pós-Operatório , Rafinose , Circulação Renal/fisiologia , Tromboxano B2/sangue , Fatores de Tempo , Doadores de TecidosRESUMO
This study investigated the effects of smoking cessation with and without nicotine substitution on the excretion of major urinary metabolites of thromboxane A2 and prostacyclin, 11-dehydrothromboxane B2 and 2,3-dinor-6-ketoprostaglandin F1alpha, respectively, as well as on the excretion of leukotriene E4 in man. Urine samples were obtained from 20 healthy non-smoking controls and from 60 healthy smoking volunteers before, and 3, 7 and 14 days after smoking cessation. Fifteen smokers quit smoking without nicotine substitution, 15 used nicotine chewing gum and 30 used nicotine patches as a substitution therapy. Urinary thiocyanate as well as cotinine and trans-3'-hydroxycotinine excretions were used as compliance and nicotine substitution indicators. 11-Dehydrothromboxane B2, 2,3-dinor-6-ketoprostaglandin F1alpha and leukotriene E4 excretion was about two, three and five times higher in smokers than in controls, respectively. Three days after smoking cessation without nicotine substitution, 11-dehydrothromboxane B2 and 2,3-dinor-6-ketoprostaglandin F1alpha levels were lowered to 75% (P<0.01) and 80% (P<0.05) of the initial values, and after 14 days to 50% (P<0.01) and 60% (P<0.05), respectively. In 3 days leukotriene E4 excretion was dropped to 70% of the initial value (P<0.05), but no further decrease was observed during the study. In individuals using nicotine chewing gum or nicotine patches no significant changes were observed in the analytes during the 2-week follow-up. The increased systemic eicosanoid synthesis observed in smokers may be involved in the harmful cardiovascular effects of smoking. The fact that eicosanoid production remains at pre-cessation level in volunteers who quit smoking but use nicotine substitution may be involved in the risk of cardiovascular complications reported during nicotine replacement therapy.