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A HPLC method with DAD detection for determination of melatonin in Lamium album flos was developed. The analysis was carried out on RP18 column using mixture of methanol and water (28:72 v/v) as a mobile phase. Established calibration curve (r>0.9994), precision (RDS values ranged from 0.5% to 1.5%), detection limit (0.025 μg/mL) and quantification limit (0.076 μg/mL) were found to be satisfactory for the proposed method. The determined content of melatonin in Lamium album flos was 0.15 μg/g of dry plant material.
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Malatonin in humans

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Melatonin, the hormone of the pineal gland, received a great deal of attention in the last decade because of its availability as over-the-counter drug or food supplement in some countries and suggested role in many vital physiological processes. Melatonin secretion is not restricted to mammals but is also produced in nonmammalian vertebrates, in some invertebrates, and in many plants, with the same molecular structure. The synthesis of melatonin is strictly controlled by lighting conditions and shows a clear circadian rhythm with low values during the daytime and significant increase at night. In this survey the basic data on melatonin significance in human physiology and in pathological processes as well as its possible thrapeutic significance are reviewed and discussed.
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Nocturnal secretion of melatonin in patients with upper digestive tract disorders

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Recently, the results of many experimental investigations have shown that melatonin possesses gastroprotective properties. On the other hand its role in pathogenesis of upper digestive tract diseases in man still remains unclear. The aim of the study was to investigate nocturnal secretion of melatonin in patients with functional and organic diseases of the upper part of digestive tract. The investigations were carried out in 149 persons, aged 21-51 years, including healthy subjects (group I, n=30), and patients with non-erosive gastroduodenal reflux (NERD, group II, n=24), with gastroesophageal reflux disease (GERD, group III, n=25), with functional dyspepsia (FD, according to the Rome III Criteria, group IV, n=36) and with recurrent duodenal ulcer (DUD, group V, n=34). Diagnoses were established on the basis of endoscopic imaging and histological examination, 24-hour pH-metry and laboratory tests. Melatonin serum concentration was measured with ELISA method. Blood samples were taken for examination in red-lighted room at 10 p.m. and on the following day at 2 and 6 a.m. The highest concentration of melatonin in all examined groups was determined at 2 a.m. The average melatonin concentration in healthy subjects was 34,7 ± 4,8 pg/ml. In patients with GERD and DUD melatonin concentration was lower than in healthy subjects - 27,2 ± 8,5 pg/ml and 25,5 ± 6,2 pg/ml respectively (p < 0,05; p < 0,01). The highest concentration of melatonin was found in patients with NERD and FD - 43,2 ± 10,8 pg/ml and 42,4 ± 10,1 pg/ml (p < 0,01; p < 0,05). The findings of this study support the notion that melatonin exerts beneficial influences on the upper digestive tract. It is likely that high or relatively correct secretion of melatonin is sufficient to prevent peptic changes in esophageal and duodenal mucosa.
Melatonin (MT) and its precursor L-tryptophan (Trp) are implicated in the protection of gastric mucosa against noxious agents. However, the role of MT and Trp on the gastric mucosal injury induced by aspirin (ASA) in human has not been investigated. Studies in animals showed that both MT and Trp given intragastrically prevents the formation of gastric mucosal lesions induced by ASA. The aim of the present study was to determine the influence of MT and Trp given orally to healthy humans on gastric mucosal lesions induced by ASA. The present study included 21 healthy, Hp-negative male volunteers with intact gastro-duodenal mucosa aging 20-50 yr. They were divided in 3 groups; group1: 7 volunteers receiving daily 2 x 1g ASA (Polfa, Rzeszow) during 11 days; group 2: 7 healthy volunteers receiving 2x1g ASA and MT (Lekam, Zakroczyn) (5 mg 30 min prior to ASA) during 11 days and group 3: 7 healthy volunteers receiving 2x1g ASA and Trp (Ardeytropin, Germany) (0.5 g 30 min prior to ASA) during 11 days. Mucosal damage was evaluated at 3rd, 7th and 11th days of ASA administration by endoscopy using Lanza score. Plasma melatonin was measured using RIA and gastric mucosal generation of PGE2 was assessed also by RIA. ASA caused marked mucosal injury at all days of its administration except day 11th when only moderate lesions were evident. Pre-treatment with MT or Trp alone was accompanied by a significant decrease in gastric mucosal lesion score. Gastric mucosal generation of PGE2 was suppressed by about 90% in subjects treated with ASA without or with MT or Trp. We concluded that: MT and its precursor Trp significantly attenuate gastric mucosal lesions induced by aspirin. The action of Trp may be be mediated by MT produced in gastrointestinal tract from Trp. The gastroprotective action of MT and Trp is independent on gastric mucosal PGE2 generation.
Assays, made on 64 mature carp females aged 5 years, were performed during the spawning season (summer) and in mid-winter. Some fish were subject to intraventricular melatonin microinjections, while other had their pineal gland excised. Intensity of fluorescence in the hypothalamic aminergic nuclei was determined with the fluorescence histochemical method. The lowest fluorescence intensity was revealed in those individuals lacking the pineal gland, the highest intensity being typical of the fish subject to intraventricular melatonin microinjections. In the winter series, all the fish showed a similar fluorescence intensity in the hypothalamic region studied. The results demonstrate a relationship between the pineal gland, melatonin, and the hypothalamic aminergic system, present in carp during the spawning period.
Melatonin, produced from L-tryptophan, protects the pancreas against acute damage by improving the antioxidative status of tissue. Melatonin receptors have been detected in the brain, but the contribution of these receptors to the pancreatic protection is unknown. The aim of our study was to compare the effects of melatonin precursor; L-tryptophan given intracerebroventricularly (i.c.v.) or intraperitoneally (i.p.) on the course of acute pancreatitis. Acute pancreatitis was induced by subcutaneous infusion of caerulein (5µg/kg-h x 5h). L-tryptophan was given i.p. (2.5, 25 or 250 mg/kg) or administered into right cerebral ventricle (0.02, 0.2 or 2.0 mg/rat) 30 min prior to the start of caerulein infusion. Plasma amylase, lipase and TNF alpha activities were measured to determine the severity of caerulein-induced pancreatitis (CIP). The lipid peroxidation products: malonylodialdehyde and 4-hydroksynonenal (MDA + 4-HNE) and activity of superoxide dismutase (SOD) were measured in the pancreas of intact or CIP rats with or without L-tryptophan pretreatment. Melatonin blood level was measured by RIA. CIP was confirmed by histological examination and manifested as an edema and rises of plasma levels of amylase, lipase and TNF alpha (by 550%, 1000% and 600%). MDA + 4-HNE was increased by 600%, whereas SOD activity was reduced by 75% in the pancreas of CIP rats. All manifestations of CIP were significantly reduced by pretreatment of the rats with L-tryptophan given i.c.v. at doses of 0.2 or 2.0 mg/rat, or by peripheral administration of this amino acid used at dose of 250 mg/kg i.p. In control rats plasma level of melatonin averaged about 40 ± 2 pg/ml and was not significantly affected by CIP, by central application of L-tryptophan (0.02, 0.2 or 2.0 mg/rat) or by peripheral administration of this melatonin precursor used at doses of 2.5 or 25 mg/kg i.p. Plasma melatonin level was markedly increased by pretreatment of the rats with L-tryptophan given i.p. at dose of 250 mg/kg. We conclude that central administration of melatonin precursor; L-tryptophan, as well as peripheral application of high dose of this melatonin precursor prevented the pancreatic damage produced by CIP. The favorable effect of peripherally administered L-tryptophan could be related to the rise of melatonin plasma level and to pancreatoprotective action of this indoleamine. The beneficial effect of centrally administered L-tryptophan could be mediated through activation of central receptors for locally produced melatonin.
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Neurokinin A and the neurohypophysial response to malatonin: in vitro studies

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The aim of this study was to investigate a possible role of neurokinin A (a member of a family of peptides known as tachykinins) in the pineal-neurohypophysial interrelationship. The effect of neurokinin A (NKA) alone or in the presence of pineal hormone - melatonin on basal and K + -stimulated vasopressin and oxytocin secretion from the hypothalamo-neurohypophysial system was studied in vitro. The present results show that NKA stimulated basal vasopressin and oxytocin release from the isolated hypothalamo-neurohypophysial system, when used at the concentration of 10-7 M/L. Melatonin diminished basal release of the neurohypophysial hormones; it also significantly inhibited the NKA-stimulated secretion of vasopressin and oxytocin. Lower concentrations of NKA did not affect the neurohypophysial hormones basal release, however, when melatonin was added to the medium enriched with NKA at the concentration of 10-9 M/L, the vasopressin secretion from the hypothalamo-neurohypophysial explants was decreased significantly. The K+ -evoked release of neurohypophysial hormones was not further modified by either NKA or melatonin. The present results confirm previous reports as to the inhibitory effect of melatonin on both vasopressin and oxytocin secretion from the hypothalamo-neurohypophysial complex in vitro. However, under present experimental conditions, the contribution of NKA in the mechanisms of pineal- neurohypophysial interrelationships has not been demonstrated.
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