Urinary 6-hydroxymelatonin Sulfate Research Articles

Melatonin: Manager of psychosomatic and metabolic disorders in polymorbid cardiovascular pathology.

ObjectivesTo investigate the relationship between changes in circadian patterns of melatonin and clinical manifestations of polymorbid cardiovascular pathology (PCVP) in young men and to analyze the effectiveness of their complex treatment.Materials and methodsWe made the immunohistochemical (IHC) analysis of epiphysis tissues from autopsies of 25 men aged 32–44 with PCVP and metabolic syndrome (MS) who had died as a result of ischemic cardiomyopathy (IC) and 25 persons after the car accident as a control group. Then, 93 young men aged 35–44 with PCVP, metabolic syndrome, and depressive spectrum disorders (DSD) were divided into three groups: (1) standard therapy; (2) standard therapy and psychotherapy sessions; (3) standard therapy in combination with psychotherapeutic and psychophysiological visual and auditory correction sessions. The control group included 24 conditionally healthy male volunteers. Before and after the treatment, we studied the anthropometric status, lipid and carbohydrate metabolism indicators, the level of urinary 6-hydroxymelatonin sulfate, the degree of nocturnal decrease in blood pressure (BP), and the relationship of these indicators with circadian variations of melatonin excretion.ResultsYoung polymorbid patients who died from IC have a lower expression of melatonin type 1 and 2 receptors. All patients with PCVP showed a decrease in the nocturnal melatonin excretion fraction and a correlation with higher severity of depressive (r = −0.72) and anxiety (r = −0.66) symptoms. Reduced values of the 6-hydroxymelatonin sulfate (6-SM) in the 1st (r = 0.45), 2nd (r = 0.39), and 3rd (r = 0.51) groups before treatment was associated with periods of increased BP. The achievement of melatonin excretion reference values and normalization of biochemical parameters of carbohydrate and lipid metabolism, daily BP profile, and psychophysiological state were noted in all three patients’ groups, with a more pronounced effect in group 3.ConclusionLow nocturnal melatonin excretion levels are associated with greater severity of clinical symptoms and a higher risk of death in patients with PCVP. Therefore, comprehensive therapy may be more effective for correcting this disease.

Eight hours of nocturnal 915 MHz radiofrequency identification (RFID) exposure reduces urinary levels of melatonin and its metabolite via pineal arylalkylamine N-acetyltransferase activity in male rats

Purpose: We investigated the effects of whole-body exposure to the 915 MHz radiofrequency identification (RFID) on mela-tonin biosynthesis and the activity of rat pineal arylalkylamine N-acetyltransferase (AANAT).Materials and methods: Rats were exposed to RFID (whole-body specific absorption rate, 4 W/kg) for 8 h/day, 5 days/week, for weeks during the nighttime. Total volume of urine excreted during a 24-h period was collected after RFID exposure. Uri-nary melatonin and 6-hydroxymelatonin sulfate (6-OHMS) was measured by gas chromatography-mass spectrometry (GC-MS) and enzyme-linked immunosorbent assay (ELISA), respectively. AANAT enzyme activity was measured using liquid biphasic dif-13 fusion assay. Protein levels and mRNA expression of AANAT was 14 measured by Western blot and reverse transcription polymerase 15 chain reaction (RT-PCR) analysis, respectively.Results: Eight hours of nocturnal RFID exposure caused a sig-nificant reduction in both urinary melatonin (p = 0. 003) and 6-OHMS (p = 0. 026). Activity, protein levels, and mRNA expres-sion of AANAT were suppressed by exposure to RFID (p < 0. 05).Conclusions: Our results suggest that nocturnal RFID exposure can cause reductions in the levels of both urinary melatonin and 6-OHMS, possibly due to decreased melatonin biosynthesis via suppression of Aanat gene transcription in the rat pineal gland.

Neuroendocrine features in extreme longevity

In order to evaluate the effects of some neuro-endocrine changes during aging we have studied adrenal, thyroid and pineal secretion in young, healthy old and centenarians. The number of subjects in each hormone group varied. The following parameters were evaluated: serum levels of cortisol, dehydroepiandrosterone-sulfate (DHEAS), free triiodothyronine (FT3), thyroxine (FT4), reverse triiodothyronine (rT3) and thyroid-stimulating hormone (TSH). Urinary 6-hydroxymelatonin sulfate (aMT6s) and free cortisol were measured twice daily. Centenarians exhibited significantly lower TSH levels together with slightly higher rT3 levels than old controls. These changes could be due to reduced 5′-deiodinase activity occurring also in absence of substantial changes of the nutritional pattern. Morning serum cortisol levels were found to be similar in the 3 age groups, whereas the decline of serum DHEAS levels was well evident also after the ninth decade of life. The cortisol/DHEAS molar ratio, which usually increases with age and considered to be an expression of a neurotoxic pattern of the steroidal milieu in the central nervous system, did not shown any further increase in centenarians. The urinary free cortisol and aMT6s excretion declined with age; however only in centenarians and in young controls aMT6s excretion was significantly higher at night than during the day. These findings suggest that the circadian rhythm of melatonin secretion is maintained in centenarians and, based on the limitations of this study, could be considered one factor in successful aging.

Physiological Significance of Cyclic Changes in Room Temperature around Dusk and Dawn for Circadian Rhythms of Core and Skin Temperature, Urinary 6-hydroxymelatonin Sulfate, and Waking Sensation just after Rising

The present study investigated whether room temperature (T(a)) cycles around dusk and dawn could influence the circadian rhythm of rectal temperature (T(core)), urinary 6-hydroxymelatonin sulfate during nocturnal sleep, and subjective assessments of sleep in humans. Six female and two male students served as participants. Two different T(a) conditions, cyclic and constant, were established. Two kinds of room temperature were provided to subjects: cyclic T(a) (gradual decrease from 27 degrees C to 24 degrees C between 1800 and 2200 h and gradual increase from 24 degrees C to 27 degrees C between 0300 and 0700 h) and constant T(a) (27 degrees C over 24 h). At cyclic T(a), the circadian nadir of T(core) rhythm was significantly advanced, while T(core) was significantly lower from 2300 to 0200 h and significantly higher from 0600 to 1000 h. The nocturnal concentration of 6-hydroxymelatonin sulfate in the urine during sleep was significantly higher during cyclic T(a). Waking sensation just after rising was significantly better with cyclic T(a). (Skin temperatures in the extremities-T(a)) were significantly higher with cyclic T(a) especially during the evening and night. Our results suggest that gradual change of room temperature in the evening and early morning is important in terms of sleep promotion and fresh awakening. It seems probable that mankind has been evolved to have deeper sleep under the influence of cyclic T(a) around dusk and dawn.

Exposure to a 50-Hz Magnetic Field Induces a Circadian Rhythm in 6-hydroxymelatonin Sulfate Excretion in Mice

The effect of magnetic field (MF) exposure on melatonin production was studied in female CD(2)F(1)(BALB/c x DBA/2) mice. The mice were exposed to a 50 Hz MF at 100 microT for 52 days and nocturnal urine was collected 1, 3, 7, 14, 16 and 23 days after the beginning of MF exposure. The animal room was illuminated for 12 h daily at 200 lux. To study the circadian rhythm of melatonin production, night and day samples of urine were collected once, at about 40 days after the beginning of MF exposure. Urinary 6-hydroxy melatonin sulfate (6-OHMS) was determined to assess melatonin production. The pineal glands were analyzed for melatonin content at the middle of the dark period. No statistically significant peak of melatonin was observed in either group. The light-regulated natural melatonin rhythm was absent in sham-exposed mice. The MF exposure caused a significant day-night difference in the 6-OHMS levels, but did not affect the total excretion of 6-OHMS during the 24-hour period. A possible interpretation of the findings is that MF exposure increases the sensitivity of the pineal gland to light in this strain normally insensitive to the circadian light variations. Further studies on interaction of light and MF exposure might help in understanding the inconsistencies of earlier research on MFs and melatonin.

Qualitative and quantitative changes of melatonin levels in physiological and pathological aging and in centenarians.

Melatonin secretion is an endogenous synchronizer, and it may possess some anti-aging properties. Thus we examined melatonin levels in physiological aging, in extreme senescence and in senile dementia. In healthy old (age 66-94 yr) and young subjects (age 23-39 yr) and in demented patients (age 68-91 yr) plasma melatonin was measured by radioimmunoassay in eight serial blood samples. In centenarians (age 100-107 yr) melatonin levels were estimated by assaying urinary 6-hydroxymelatonin sulfate (aMT6s) in two different urine samples collected from 08:00 to 20:00 hours and from 20:00 to 08:00 hours. These data were compared with the aMT6s excretion of old and young controls. Elderly subjects, demented or not, exhibited a flattened circadian profile of plasma melatonin, because of the suppression of the nocturnal peak. An age-related decline of the circadian amplitude of the melatonin rhythm occurred in old subjects, especially in demented individuals. Furthermore, the melatonin nocturnal peak was significantly correlated with the severity of the cognitive impairment. aMT6s urinary excretion also declined with age. However, as in young controls, in centenarians the aMT6s excretion was significantly higher at night than during the day. In conclusion, pineal melatonin secretion is affected by age and by the degree of cognitive impairment. In centenarians the maintenance of the circadian organization of melatonin secretion may suggest that the amplitude of the nocturnal peak and/or the persistence of a prevalent nocturnal secretion may be an important marker of biological age and of health status.

A naturally occurring −263G/C variant of the human AA-NAT gene and overnight melatonin production

Several lines of evidence show that the daily amount of melatonin produced differs greatly between individuals. Any polymorphism in the gene of arylalkylamine N-acetyltransferase (AA-NAT), a critical enzyme involved in melatonin biosynthesis, may contribute to the variability of melatonin production. The present study investigated the possible association between overnight melatonin excretion and a commonly occurring −263G/C polymorphism in the promoter region of the human AA-NAT gene. However, we found that −263G/C variant had no effect on the overnight 6-OHMS excretion. In this study, individual genotyping for −263G/C was determined by denaturing high performance liquid chromatography (DHPLC) and confirmed by sequencing. The overnight urinary 6-hydroxymelatonin sulfate (6-OHMS) excretion was quantified by enzyme-linked immunosorbent assay (ELISA).

Melatonin production during childhood and adolescence: a longitudinal study on the excretion of urinary 6-hydroxymelatonin sulfate.

Cross-sectional data on urinary 6-hydroxymelatonin sulfate (6-OHMS) excretion in children suggest a constant melatonin secretion during growth. The present longitudinal study concerned, accordingly, the intra-individual stability of melatonin production during childhood and adolescence. Urine samples collected during a longitudinal investigation of healthy white children and adolescents were analyzed. Forty-six boys and 38 girls were chosen for the present study. They had passed 3-15 annual examinations between their 3rd and 18th yr of age. Each examination included the collection of urine over 24 hr. The daily urinary output of 6-OHMS of the overall 621 samples was quantified by enzyme-linked immunosorbent assay. The analyses clearly revealed for the first time that, despite huge inter-individual differences, melatonin production remains constant in one and the same individual during childhood and adolescence. Additionally, neither a significant sex difference was observed nor was the 6-OHMS output affected by season. The dramatic decrease of plasma melatonin levels as described in the literature is mainly related to an increase in body size rather than to decreasing pineal secretion.

Nocturnal 6-hydroxymelatonin sulfate excretion in female workers exposed to magnetic fields.

The objective of this study was to determine whether daytime occupational exposure to extremely low frequency magnetic fields (MFs) suppresses nocturnal melatonin production. Sixty female volunteers were recruited. Thirty-nine worked in a garment factory, and 21 office workers served as a reference group. Exposure assessment was based on the type of sewing machine used and MF measurements around each type of machine. Eye-level MF flux density was used to classify the operators to higher (>1 microT) and lower (0.3-1 microT) exposure categories. A third group of factory workers had diverse MF exposures from other sources. The reference group had average exposure of about 0.15 microT. Urine samples were collected on Friday and Monday for three consecutive weeks. Melatonin production was assessed as urinary 6-hydroxymelatonin sulfate (6-OHMS) excretion. The ratio of Friday morning/Monday morning 6-OHMS was used to test the hypothesis that melatonin production is suppressed after 4 days of occupational MF exposure with significant recovery during the weekend. Possible chronic suppression of melatonin production was evaluated by studying exposure-related differences in the Friday values by multivariate regression analysis. The Monday/Friday ratios were close to 1.0, suggesting that there is no increase in melatonin production over the weekend. The average 6-OHMS excretion on Friday was lower among the factory workers than in the reference group, but no monotonous dose-response was observed. Multivariate regression analysis identified MF exposure, smoking, and age as significant explanatory variables associated with decreased 6-OHMS excretion.

Twenty‐four hour urinary excretion of 6‐hydroxymelatonin sulfate in Down syndrome subjects

Because of the overexpression of the enzyme superoxide dismutase, individuals with Down syndrome (DS) are believed to suffer from increased oxidative stress as a result of the excessive production of oxygen-based free radicals; their exposure to higher than normal free radical production may account in part for signs of premature aging, early onset of cataracts, and of Alzheimer's disease. Free radicals are normally neutralized by free radical scavengers and other antioxidants. The pineal hormone melatonin is a potent scavenger of both the hydroxyl and peroxyl radicals, both of which are highly toxic, and a stimulator of the antioxidative enzyme glutathione peroxidase. Considering this, we deemed it important to define the day/night rhythm and levels of melatonin production in DS subjects. To do this, we assessed the urinary excretion of the chief melatonin metabolite, 6-hydroxymelatonin sulfate, throughout a 24 hr period in DS subjects; comparisons were made with the metabolite levels in the urine of non-Down siblings and parents of the DS subjects. All 8 non-Down subjects exhibited what was classified as normal urinary excretion of 6-hydroxymelatonin sulfate with the usual low daytime and high night-time levels of the melatonin metabolite. Of 12 DS subjects studied, 10 exhibited the normal day/night rhythm in urinary 6-hydroxymelatonin sulfate levels; 2 subjects were devoid of a rhythm. However, when all the data from each group were averaged, there were no noticeable differences in the absolute levels or 24 hr variations in urinary 6-hydroxymelatonin sulfate excretion between DS and non-Down subjects.

Urinary 6-hydroxymelatonin sulfate as a measure of melatonin secretion during acute tryptophan depletion

The essential amino acid tryptophan (TRP) is the precursor for serotonin (5-HT), which is in turn an intermediary product in the synthesis of melatonin. Acute TRP depletion has recently been shown to decrease nocturnal plasma levels of melatonin in humans. Melatonin is metabolized to 6-hydroxymelatonin sulfate (6-SM), a highly stable end-product which is excreted into urine. To determine the effects of TRP bioavailability on 6-SM, 11 healthy volunteers underwent active and sham TRP depletion in a randomized, double-blind fashion. Samples of plasma free and total TRP, plasma melatonin, and urinary 6-SM were obtained before and after the depletion. Acute TRP depletion decreased free and total plasma tryptophan levels by more than 80% from baseline levels. Nocturnal 6-SM excretion was significantly decreased and highly correlated with decreases in plasma melatonin. These results suggest that nocturnal urinary 6-SM excretion is a valid measure of melatonin secretion under conditions of decreased 5-HT function. Collection of urine for 6-SM is considerably easier than nocturnal plasma sampling for melatonin. Further studies are needed to clarify the relationship between 6-SM excretion and other measures of 5-HT function in neuropsychiatric disorders.

Evidence for an Effect of ELF Electromagnetic Fields on Human Pineal Gland Function

A study was carried out to determine possible effects of 60-Hz electromagnetic-field exposure on pineal gland function in humans. Overnight excretion of urinary 6-hydroxymelatonin sulfate (6-OHMS), a stable urinary metabolite of the pineal hormone melatonin, was used to assess pineal gland function in 42 volunteers who used standard (conventional) or modified continuous polymer wire (CPW) electric blankets for approximately 8 weeks. Volunteers using conventional electric blankets showed no variations in 6-OHMS excretion as either a group or individuals during the study period. Serving as their own controls, 7 of 28 volunteers using the CPW blankets showed statistically significant changes in their mean nighttime 6-OHMS excretion. The CPW blankets switched on and off approximately twice as often when in service and produced magnetic fields that were 50% stronger than those from the conventional electric blankets. On the basis of these findings, we hypothesize that periodic exposure to pulsed DC or extremely low frequency electric or magnetic fields of sufficient intensity and duration can affect pineal gland function in certain individuals.