RESUMO
PURPOSE: Changes in the circadian rhythm may contribute to the development of cancer and are correlated with the high risk of breast cancer (BC) in night workers. Melatonin is a hormone synthesized by the pineal gland at night in the absence of light. Levels of melatonin and the metabolite of oxidative metabolism AFMK (acetyl-N-formyl-5-methoxykynurenamine), are suggested as potential biomarkers of BC risk. The aims of this study were to evaluate levels of melatonin and AFMK in women recently diagnosed with BC, women under adjuvant chemotherapy, and night-shift nurses, and compare them with healthy women to evaluate the relation of these compounds with BC risk. METHODS: Blood samples were collected from 47 women with BC, 9 healthy women, 10 healthy night shift nurses, and 6 patients under adjuvant chemotherapy. Compound levels were measured by mass spectrometry. RESULTS AND CONCLUSIONS: Our results showed that women with BC had lower levels of melatonin compared to control group women, and even lower in night-shift nurses and in patients under adjuvant chemotherapy. There was no significant difference of AFMK levels between the groups. In addition to this, high levels of melatonin and AFMK were related to patients with metastasis, and high levels of AFMK were related to the presence of lymph node-positive, tumor > 20 mm and patients who sleep with light at night. Our results showed a reduction of melatonin levels in BC patients, suggesting a relation with the disease, and in addition, point to the importance of melatonin supplementation in women that work at night to reduce the BC risk.
Assuntos
Neoplasias da Mama/sangue , Cinuramina/análogos & derivados , Melatonina/sangue , Biomarcadores/sangue , Ritmo Circadiano/fisiologia , Feminino , Humanos , Cinuramina/sangue , Metástase Linfática , Pessoa de Meia-Idade , PrognósticoRESUMO
Melatonin (N-acetyl-5-methoxytryptamine) is implicated in physiologic changes related to light-dark cycles and has been recently found to display antioxidant properties. It is known that the reaction of melatonin with certain reactive oxygen and nitrogen species, such as hydrogen peroxide and singlet oxygen, produces N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK). We report herein on the development of a new liquid chromatography/tandem mass spectrometry (LC/ESI/MS-MS) assay to quantitatively determine melatonin and AFMK. The stable isotopic internal standard of melatonin-D3 was synthesized by the reaction of 5-methoxytryptamine with deuterated acetyl chloride (CD3COCl). Labeled AFMK (AFMK-D3) was obtained after photooxidation of melatonin-D3. The predominant ion [M + H]+ in the full scan mass spectra of melatonin, melatonin-D3, AFMK and AFMK-D3 were located, respectively, at m/z = 233, 236, 265 and 268. The collision-induced dissociation of the molecules revealed a predominant fragment at m/z = 174 for melatonin and melatonin-D3 (loss of the N-acetyl group), and at m/z = 178 for AFMK and AFMK-D3 (loss of both the N-acetyl and the N-formyl groups). The m/z transitions from 233 to 174 (melatonin), from 236 to 174 (melatonin-D3), from 265 to 178 (AFMK), and from 268 to 178 (AFMK-D3) were therefore chosen for the multiple reaction monitoring detection experiments, ensuring a high specificity and an accurate quantification of melatonin and AFMK in human plasma.
Assuntos
Cromatografia Líquida/métodos , Cinuramina/análogos & derivados , Cinuramina/sangue , Cinuramina/síntese química , Melatonina/sangue , Melatonina/síntese química , Espectrometria de Massas por Ionização por Electrospray/métodos , Bioquímica/métodos , Calibragem , Cromatografia Líquida/normas , Humanos , Espectrometria de Massas por Ionização por Electrospray/normasRESUMO
1. The present study was undertaken to determine if the platelet monoamine oxidase (MAO) activity of children with childhood-onset Pervasive Developmental Disorders (PDD), atypical PDD and autistic children differs from MAO of normal children of the same age. 2. The kinetic parameters of MAO activity (Km and Vmax for kynuramine as substrate in 100 mM sodium phosphate buffer, pH 7.4 at 37 degrees C) were determined for platelets from autistic (N = 6), childhood onset PDD (N = 6) and atypical PDD (N = 6) children and 14 controls aged 6-10 years. 3. PDD children had significantly lower Km (4.41 +/- 0.26 vs 5.30 +/- 0.23 microM) and Vmax (16.77 +/- 1.56 vs 22.15 +/- 2.16 nmol h-1 mg protein-1) than control children. The reduction in Vmax was demonstrable in MAO activity measured with 100 microM substrate (14.93 +/- 1.13 vs 20.96 +/- 2.10 nmol h-1 mg-1). 4. These data show that childhood-onset PDD patients, in which the syndrome was complete, presented the lowest levels of platelet MAO activity.