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Showing papers in "Journal of Pineal Research in 1999"


Journal ArticleDOI
TL;DR: The pineal secretory product melatonin reportedly regulates release of growth hormone in humans and prevents phototherapy-induced hypocalcemia in newborn rats, suggesting that melatonin affects bone metabolism as mentioned in this paper.
Abstract: The pineal secretory product melatonin reportedly regulates release of growth hormone in humans and prevents phototherapy-induced hypocalcemia in newborn rats, suggesting that melatonin affects bone metabolism. Little is known about the effects of melatonin on bone in vitro or in vivo. The present study was undertaken to examine whether melatonin acts directly on normal human bone cells (HOB-M cells) and human osteoblastic cell line (SV-HFO cells) to affect osteogenic action in vitro. The effect of melatonin on bone cell proliferation was determined using the 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carbo xanilide (XTT) assay after a 24 hr incubation with melatonin. Melatonin significantly and dose-dependently increased the proliferation in HOB-M cells and SV-HFO cells by 215 +/- 22.1%, and 193 +/- 6.4%), respectively, with a maximal effect at a concentration of 50 microM. To evaluate the effect of melatonin on bone cell differentiation, alkaline phosphatase (ALP) activity, osteocalcin secretion and procollagen type I c-peptide (PICP) production (a measure of type I collagen synthesis) were measured after a 48 hr treatment. While melatonin at micromolar concentrations did not significantly affect either the ALP activity or the osteocalcin secretion, it significantly and dose-dependently increased the PICP production in HOB-M cells and SV-HFO cells by 983 +/- 42.2%, and 139 +/- 4.2%, respectively, with the maximal stimulatory doses between 50 and 100 microM. These results provide new evidence that melatonin stimulates the proliferation and type I collagen synthesis in human bone cells in vitro, suggesting that melatonin may act to stimulate bone formation.

209 citations


Journal ArticleDOI
TL;DR: Results showed that both melatonin and TAS in human serum exhibited 24 hr variations with nocturnal peak values at 01:00 hr, and suggested that melatonin contributes to the total antioxidative capability of human serum.
Abstract: This work evaluates whether physiological concentrations of the pineal secretory product melatonin contribute to the total antioxidant status (TAS) of human serum. Day and nighttime serum samples were collected from healthy volunteers ranging from 2 to 89 years of age and used to measure melatonin and TAS. Results showed that both melatonin and TAS in human serum exhibited 24 hr variations with nocturnal peak values at 01:00 hr. Moreover, exposure of volunteers to light at night resulted in clear decreases of both TAS and melatonin. Furthermore, when melatonin was removed from sera collected at night, the TAS value of the sample was reduced to basal daytime values. In aging studies, it was found that nocturnal serum values of TAS and melatonin exhibited maximal values during the first four decades; thereafter, these values decreased as age advanced. In 60-year-old individuals, day/night differences in serum melatonin and TAS levels were clearly diminished, by more than 80%, with these differences being completely abolished in older individuals. Our results suggest that melatonin contributes to the total antioxidative capability of human serum. This antioxidant contribution of melatonin is reduced as age advances correlating with the age-related reduction of melatonin.

186 citations


Journal ArticleDOI
TL;DR: Melatonin, which has a methoxy group in the 5‐position and an acetyl side chain, exhibited the most potent scavenging activity among the compounds tested, while Serotonin and N‐acetyl‐5 hydroxytryptamine, and 5‐hydroxytryptophan showed moderate scavengingActivity compared to melatonin.
Abstract: Nitric oxide (NO) scavenging activity of melatonin, N-acetyl-5-hydroxytryptamine, serotonin, 5-hydroxytryptophan and L-tryptophan was examined by the Griess reaction using flow injection analysis. 1-Hydroxy-2-oxo-3-(N-methyl-3-aminopropyl)-3-methyl-1-triazene (NOC-7) was used as NO generator. The Griess reagent stoichiometrically reacts with NO2-, which was converted by a cadmium-copper reduction column from the stable end products of NO oxidation. Except for tryptophan, all the compounds examined scavenged NO in a dose-dependent manner. Melatonin, which has a methoxy group in the 5-position and an acetyl side chain, exhibited the most potent scavenging activity among the compounds tested. Serotonin, N-acetyl-5-hydroxytryptamine, and 5-hydroxytryptophan, respectively, showed moderate scavenging activity compared to melatonin. Tryptophan, which has neither a methoxy nor a hydroxyl group in the 5-position, exhibited the least NO scavenging activity.

182 citations


Journal ArticleDOI
TL;DR: Based on the published data, it is currently unclear if EMF and electric light exposure are significant risk factors for breast cancer, but further study appears warranted.
Abstract: Worldwide, breast cancer is the most common malignancy accounting for 20-32% of all female cancers. This review summarizes the peer-reviewed, published data pertinent to the hypothesis that increased breast cancer in industrialized countries is related to the increased use of electricity [Stevens, R.G., S. Davis 1996]. That hypothesis specifically proposes that increased exposure to light at night and electromagnetic fields (EMF) reduce melatonin production. Because some studies have shown that melatonin suppresses mammary tumorigenesis in rats and blocks estrogen-induced proliferation of human breast cancer cells in vitro, it is reasoned that decreased melatonin production leads to increased risk of breast cancer. To evaluate this hypothesis, the paper reviews epidemiological data on associations between electricity and breast cancer, and assesses the data on the effects of EMF exposure on melatonin physiology in both laboratory animals and humans. In addition, the results on the effects of melatonin on in vivo carcinogenesis in animals are detailed along with the controlled in vitro studies on melatonin's effects on human breast cancer cell lines. The literature is evaluated for strength of evidence, inter-relationships between various lines of evidence, and gaps in our knowledge. Based on the published data, it is currently unclear if EMF and electric light exposure are significant risk factors for breast cancer, but further study appears warranted. Given the ubiquitous nature of EMF and artificial light exposure along with the high incidence of breast cancer, even a small risk would have a substantial public health impact.

129 citations


Journal ArticleDOI
TL;DR: It is concluded that melatonin production is lower in older people, but that the change occurs very early in life, around 20–30 yr of age.
Abstract: The apparent age-related decline in melatonin production has been thought to continue in a secular manner across the lifespan. While it is clear that melatonin levels in children and adolescents are elevated compared to older individuals, the question of whether there is a sudden or gradual change has not been adequately addressed. In this study, we report the excretion of the melatonin metabolite, 6-sulfatoxymelatonin in 253 subjects aged between 21 and 82 yr. The correlation with age was significant (r = -0.24; P < 0.05). When the data was analysed by ANOVA using 5-yr age spans, there was a significant effect of age, but post hoc analysis indicated that after 25 yr of age there was no significant decline in excretion of the metabolite. Thus, although the oldest subjects excreted 36% less melatonin metabolite than the youngest, the decrease occurred at a very early age. In the second part of the study, we re-evaluated the data from seven previous studies that measured plasma melatonin levels or metabolite excretion across a wide range of ages and 11 studies comparing young versus older subjects. Statistical analysis by ANOVA again suggested that the changes in melatonin occurring with age were essentially complete before 30 yr of age. The youngest subjects produced at the most twice the amount of melatonin as the oldest subjects. Finally, we evaluated the mean plasma melatonin levels in 144 groups of normal subjects reported in 137 separate publications with respect to age. Again, whereas there was a significant correlation with age, ANOVA showed that there was no difference between groups after 35 yr of age, and the oldest groups had levels that were only 43% of the youngest groups. We conclude that melatonin production is lower in older people, but that the change occurs very early in life, around 20-30 yr of age.

126 citations


Journal ArticleDOI
TL;DR: Under conditions where cellular homogenates are used, vitamin E and the three pineal indoleamines protected the retinal cells from NO‐induced LPO damage and vitamin E acted synergistically with indoleamine in combating oxidative retinal damage.
Abstract: Oxidative damage to retinal cell membranes can lead to sight-threatening ocular diseases. Pineal indoleamines are naturally located and synthesized in the retina, and they possibly protect the retina from oxidative cell damage. In this study, we compared the efficacy of three different pineal indoleamines (melatonin, N-acetylserotonin, and pinoline) with vitamin E, a well-known antioxidant, against nitric oxide (NO)-induced lipid peroxidation (LPO) in rat retinal homogenates. The possible synergistic effect of these agents was also studied. Retinal homogenates were incubated with sodium nitroprusside, which releases NO'. The LPO product, malondialdehyde (MDA), provided an index of cell damage. The results show that vitamin E and indoleamines significantly reduced MDA levels in a dose-dependent manner. When vitamin E was combined with the indoleamines, the protection was synergistically enhanced. In summary, under conditions where cellular homogenates are used (a) vitamin E and the three pineal indoleamines protected the retinal cells from NO-induced LPO damage; (b) the efficacies of each of these compounds had the following relationships: vitamin E > N-acetylserotonin > pinoline > melatonin; (c) vitamin E acted synergistically with indoleamines in combating oxidative retinal damage. Whether these same associations would exist in vivo after treatment with these compounds is unknown. The pharmacological potential of indoleamines, possibly in combination with vitamin E, in preventing retinal pathogenesis deserves further investigation.

119 citations


Journal ArticleDOI
TL;DR: Findings may suggest that the molecule could easily scavenge aqueous as well as lipophilic radicals.
Abstract: The interaction of melatonin with water containing either sodium bis (2-ethylhexyl) sulfosuccinate (AOT) or soybean phosphatidylcholine (lecithin) reversed micelles has been investigated by UV absorption spectroscopy, at a molar ratio of melatonin:surfactant 1:800 for AOT and 1:400 for lecithin reversed micelles, and by varying the water:surfactant molar ratio (R). Our results suggest that in the presence of domains from apolar organic solvent to surfactant and to water, melatonin positions itself in the micellar phase, with a preferential location in the surfactant polar head group domain, independent of the nature of the surfactant and the amount of water encapsulated into the micellar core. Effects are due to the hydrophilic and lipophilic moieties of melatonin. The effectiveness of melatonin as an electron donor and free radical scavenger has been recently recognized. While supporting the hypothesis that melatonin may provide antioxidant protection without the benefit of receptors, present findings may suggest that the molecule could easily scavenge aqueous as well as lipophilic radicals.

113 citations


Journal ArticleDOI
TL;DR: Results indicate that melatonin administration to the pregnant rat may prevent the ischemia/reperfusion‐induced oxidative lipid and DNA damage in fetal rat brain.
Abstract: To investigate whether melatonin reduces the susceptibility of the fetal rat brain to oxidative damage of lipids and DNA, we created a model of fetal ischemia/reperfusion using rats at day 19 of pregnancy. Fetal ischemia was induced by bilateral occlusion of the utero-ovarian artery for 20 min. Reperfusion was achieved by releasing the occlusion and restoring the circulation for 30 min. A sham operation was performed in control rats. Melatonin (10 mg/kg) or vehicle was injected intraperitoneally 60 min prior to the occlusion. We measured the concentration of thiobarbituric acid reactive substances (TBARS) in fetal brain homogenates, as well as levels of deoxyguanosine (dG) and 8-hydroxydeoxyguanosine (8-OHdG) in DNA extracted from those homogenates. Ischemia for 20 min did not significantly alter the levels of dG, 8-OHdG, and TBARS. Subsequent reperfusion, however, led to a significant reduction in the dG level (P < 0.05) and to significant increases in the levels of 8-OHdG (P < 0.05) and TBARS (P < 0.05), and in the 8-OHdG/dG ratio (P < 0.005). Melatonin administration prior to ischemia significantly reduced the ischemia/reperfusion-induced increases in the levels of 8-OHdG (14.33 +/- 6.52-5.15 +/- 3.28 pmol/mg of DNA, P < 0.001) and TBARS (11.61 +/- 3.85-4.73 +/- 3.80 nmol/mg of protein, P < 0.001) as well as in the 8-OHdG/dG ratio (7.19 +/- 2.49-1.61 +/- 0.98, P < 0.001). Furthermore, melatonin significantly increased the dG level (210.19 +/- 49.02-299.33 +/- 65.08 nmol/mg of DNA, P < 0.05). Results indicate that melatonin administration to the pregnant rat may prevent the ischemia/reperfusion-induced oxidative lipid and DNA damage in fetal rat brain.

109 citations


Journal ArticleDOI
TL;DR: The data indicate that melatonin has the ability to protect the genetic material of hematopoietic cells of mice from the damaging effects of acute whole‐body irradiation.
Abstract: The objective of this study was to examine the potential radioprotective properties of pharmacological doses of melatonin in whole-body irradiated mice. CD2-F1 male mice were treated with melatonin, a secretory product of the pineal gland, and then whole-body irradiated with an acute dose (150 cGy) of 137Cs gamma rays. Peripheral blood and bone marrow cells were examined for genetic damage, which was determined by comparing the incidence of micronuclei (MN) in both melatonin pre-treated and non-treated irradiated animals (and control mice). The percentages of polychromatic erythrocytes (PCEs) in unirradiated mice ranged between 3.1 +/- 0.23 and 3.2 +/- 0.19 in the peripheral blood and between 51.0 +/- 2.03 and 52.8 +/- 2.00 in the bone marrow. Whole-body irradiation resulted in a significant decrease in the percentages of PCEs in the peripheral blood and bone marrow cells. In both tissues, irradiated mice that were pre-treated with melatonin (5 or 10 mg/kg) exhibited a dose-dependent increase in the observed incidence of PCEs relative to the expected incidence. The incidence of MN in unirradiated mice ranged between 4.2 +/- 0.92 and 4.6 +/- 0.97 in the peripheral blood and between 5.0 +/- 1.05 and 5.5 +/- 1.08 in the bone marrow. Whole-body irradiation resulted in a significant increase in the incidence of MN in both tissues. In both tissues, irradiated mice that were pre-treated with melatonin exhibited a significant and dose-dependent reduction in the observed incidence of MN (relative to the expected incidence). Under the experimental conditions tested, the data indicate that melatonin has the ability to protect the genetic material of hematopoietic cells of mice from the damaging effects of acute whole-body irradiation.

107 citations


Journal ArticleDOI
TL;DR: In an ovarian adenocarcinoma cell line (BG‐1), it is found that melatonin at concentrations of 10−9‐10−7 M caused a 20–25% reduction in cell number, but the effect is small and the results are inconsistent.
Abstract: Melatonin is reported to reduce proliferation in many cell types, but the effect is small and the results are inconsistent. Information on the mechanism by which melatonin exerts its antiproliferative effects might provide insight into the variability of the response. In an ovarian adenocarcinoma cell line (BG-1), we find that melatonin at concentrations of 10(-9)-10(-7) M caused a 20-25% reduction in cell number. Melatonin also resulted in a similar reduction in [3H]-thymidine incorporation with no significant increase in cell death as measured by trypan blue incorporation. The Kd for melatonin reduction in cell number was approximately 5 x 10(-10) M. Melatonin ML2 receptors have a Kd for melatonin binding in the low nM range and are linked to the production of the calcium mobilizing agent inositol-1,4,5-trisphosphate (IP3). To investigate whether melatonin signaling involves an increase in cytosolic-free calcium. BG-1 cells were loaded with the calcium sensitive indicator, fura-2. Acute addition of melatonin (10(-5)-10(-9) M) did not alter cytosolic calcium. Addition of the putative nuclear receptor agonist CGP52608 caused a dose-dependent inhibition of cell number with a Kd of approximately 2 x 10(-9) M. Addition of CGP52608 caused a similar reduction in [3H]-thymidine incorporation. Neither melatonin (10(-8) M-10(-5) M) nor CGP52608 at concentrations below 10(-7) M induced cell death associated with the inhibition of cell proliferation; however, addition of CGP52608 at a high dose (10(-7) M) caused an increase in cell death, consistent with apoptosis. Growth inhibition by melatonin or CGP52608 did not alter the percentage of cells in G1 versus S/G2/M.

107 citations


Journal ArticleDOI
TL;DR: The species‐specific levels and a distinct distribution of melatonin in the layers of the digestive tube indicates that this indole may be involved in the modulation of gastrointestinal function of monogastric as well as polygastric ungulates, albeit in a different capacity.
Abstract: Melatonin concentrations were measured in serum, luminal fluid, and tissues of the mucosa and muscularis of the entire bovine and porcine gastrointestinal tract (GIT). In both species, GIT levels profoundly exceeded serum levels. In pigs, melatonin was lowest in the luminal fluid and highest in the mucosa. No difference was found in various layers of bovine GIT. Compared to pigs, cows had higher melatonin levels in the stomach and ileum, but lower in the cecum and colon. There was no difference in melatonin levels between anterior and posterior segments of bovine GIT, whereas pigs exhibited several fold higher concentration of melatonin in the posterior segment (cecum and colon). Conversely, melatonin values in the anterior segment were significantly higher in cows, but in the posterior segments porcine values were higher. In cows, concentrations in the mucosa correlated with levels in the muscularis. Melatonin levels in the mucosa and muscularis were higher in the rumen and reticulum than in the omasum and abomasum. The species-specific levels and a distinct distribution of melatonin in the layers of the digestive tube indicates that this indole may be involved in the modulation of gastrointestinal function of monogastric as well as polygastric ungulates, albeit in a different capacity.

Journal ArticleDOI
TL;DR: The results indicate that melatonin is actively taken up into crythrocytes under oxidative stress, and is consumed in the defence of the cell, delaying Hb denaturation and release of hemin.
Abstract: Antioxidant activity of melatonin in human erythrocytes, exposed to oxidative stress by cumene hydroperoxide (cumOOH), was investigated. CumOOH at 300 microM progressively oxidized a 1% suspension of red blood cells (RBCs), leading to 100% hemolysis in 180 min. Malondialdehyde and protein carbonyls in the membrane showed a progressive increase, as a result of the oxidative damage to membrane lipids and proteins, reaching peak values after 30 and 40 min, respectively. The membrane antioxidant vitamin E and the cytosolic reduced glutathione (GSH) were totally depleted in 20 min. As a consequence of the irreversible oxidative damage to hemoglobin (Hb), hemin accumulated into the RBC membrane during 40 min. Sodium dodecyl sulfate (SDS) gel electrophoresis of membrane proteins showed a progressive loss of the cytoskeleton proteins and formation of low molecular weight bands and protein aggregates, with an increment of the intensity of the Hb band. Melatonin at 50 microM strongly enhanced the RBC resistance to oxidative lysis, leading to a 100% hemolysis in 330 min. Melatonin had no effect on the membrane lipid peroxidation, nor prevented the consumption of glutathione (GSH) or vitamin E. However, it completely inhibited the formation of membrane protein carbonyls for 20 min and hemin precipitation for 10 min. The electrophoretic pattern provided further evidence that melatonin delayed modifications to the membrane proteins and to Hb. In addition, RBCs incubated for 15 min with 300 microM cumOOH in the presence of 50 microM melatonin were less susceptible, when submitted to osmotic lysis, than cells incubated in its absence. Extraction and high-performance liquid chromatography (HPLC) analysis showed a much more rapid consumption of melatonin during the first 10 min of incubation, then melatonin slowly decreased up to 30 min and remained stable thereafter. Equilibrium partition experiments showed that 15% of the melatonin in the incubation mixture was recovered in the RBC cytosol, and no melatonin was extracted from RBC membrane. However, 35% of the added melatonin was consumed during RBC oxidation. Hydroxyl radical trapping agents, such as dimethylsulfoxide or mannitol, added into the assay in a 1,000 times molar excess, did not vary melatonin consumption, suggesting that hydroxyl radicals were not involved in the indole consumption. Our results indicate that melatonin is actively taken up into erythrocytes under oxidative stress, and is consumed in the defence of the cell, delaying Hb denaturation and release of hemin. RBCs are highly exposed to oxygen and can be a site for radical formation, under pathological conditions, which results in their destruction. A protective role of melatonin should be explored in hemolytic diseases.

Journal ArticleDOI
TL;DR: Previous findings with Aβ25‐35 are supported, including a causative role for Aβ in the mitochondrial oxidative lesions present in AD brains, and the neuroprotective role of melatonin in Aβ‐mediated oxidative injury is confirmed.
Abstract: Most contemporary progress in Alzheimer's disease (AD) stems from the study of a 42 43 amino acid peptide. called the amyloid beta protein (Abeta), as the main neuropathologic marker of the disorder. It has been demonstrated that Abeta has neurotoxic properties and that such effects are mediated by free-radicals. Exposure of neuronal cells to Abeta results in a spectrum of oxidative lesions that are profoundly harmful to neuronal homeostasis. We had previously shown that Abeta25-35 induces oxidative damage to mitochondrial DNA (mtDNA) and that this modality of injury is prevented by melatonin. Because Abeta25 35 does not occur in AD and because the mode of toxicity by Abeta25-35 may be different from that of Abeta1-42 (the physiologically relevant form of Abeta), we extended our initial observations to determine whether oxidative damage to mtDNA could also be induced by Abeta1-42 and whether this type of injury is prevented by melatonin. Exposure of human neuroblastoma cells to Abeta1-42 resulted in marked oxidative damage to mtDNA as determined by a quantitative polymerase chain reaction method. Addition of melatonin to cell cultures along with Abeta completely prevented the damage. This study supports previous findings with Abeta25-35, including a causative role for Abeta in the mitochondrial oxidative lesions present in AD brains. Most important, the data confirms the neuroprotective role of melatonin in Abeta-mediated oxidative injury. Because melatonin also inhibits amyloid aggregation, lacks toxicity, and efficiently crosses the blood-brain barrier, this hormone appears superior to other available antioxidants as a candidate for pharmacologic intervention in AD.

Journal ArticleDOI
TL;DR: Melatonin at picomolar and low nanomolar concentrations was determined to be 100 times more potent in inhibiting the light‐induced oxidative processes than was vitamin E and both compounds exerted potent prooxidant effects at micromolar concentrations that is above the physiological levels of melatonin.
Abstract: Dark-adapted, single photoreceptors isolated from the frog retina produce reactive oxygen species (ROS) after about 1 min of illumination with saturating light that we verified by their oxidation of preloaded dihydrorhodamine 123 (DHR) into the fluorescent rhodamine 123 (RHO). In this preparation we tested the antioxidant effects of vitamin E and of melatonin. Melatonin at picomolar and low nanomolar concentrations was determined to be 100 times more potent in inhibiting the light-induced oxidative processes than was vitamin E. On the contrary, both compounds exerted potent prooxidant effects at micromolar concentrations that is above the physiological levels of melatonin. This provides evidence that physiological concentrations of melatonin in a living cell may exert protective actions against a natural oxidant stimulus (light). This helps to define the functional role of endogenous melatonin in photoreceptors, which by their physiological characteristics, are among the marked producers of ROS in the organism.

Journal ArticleDOI
TL;DR: The data suggest that the reported anti‐proliferative action of melatonin on human choriocarcinoma JAr cells may be mediated, in part, by MT2 melatonin receptor.
Abstract: Melatonin, the pineal neurohormone, is an evolutionarily conserved photoperiodic signaling molecule with diverse functions that include the entrainment of human circadian rhythms Although evidence supporting a direct inhibitory action of melatonin on human cancer cell proliferation exists in the literature, the molecular and cellular signaling mechanisms involved are largely undefined In our study, significant inhibition of human choriocarcinoma JAr cell proliferation at physiological and pharmacological concentrations of melatonin was observed 2-Iodomelatonin, a high affinity melatonin receptor agonist, was more potent than melatonin in inhibiting JAr cell proliferation In addition, the presence of putative melatonin receptors in choriocarcinoma was suggested by the demonstration of specific 2-[125I]iodomelatonin binding to the tumor Interestingly, the selective MT2 melatonin receptor ligand, 4-phenyl-2-propionamidotetraline (4-P-PDOT), was found to exert not only concentration-dependent anti-proliferative actions on JAr cells, but also additive effects with melatonin in inhibiting JAr cell proliferation Furthermore, MT2 melatonin receptor gene expression by JAr cells was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization (ISH) Taken together, our data suggest that the reported anti-proliferative action of melatonin on human choriocarcinoma JAr cells may be mediated, in part, by MT2 melatonin receptor Moreover, analysis of melatonin effect on cell cycle kinetics indicated that G1/S transition delay may underlie the observed inhibition of choriocarcinoma cell proliferation by melatonin

Journal ArticleDOI
TL;DR: It is suggested that melatonin probably affects the secretion of sAPP in the conditioned medium by interfering with its full maturation, and melatonin also affects the presysnaptic terminal marker.
Abstract: Melatonin is released in mammals during the dark phase of the circadian cycle, and its production declines with age in animals and humans. Since supplemental administration of melatonin may be beneficial in delaying age-related degenerative conditions, it is necessary to study its effect on neuronal differentiation and the processing of key neuronal proteins, such as beta-amyloid precursor protein (beta APP) and synaptophysin. One of the important pathological hallmarks of Alzheimer's disease (AD) is the cerebrovascular deposition of amyloid plaques. The amyloid in senile plaques is mainly composed of the amyloid beta-peptide (A beta) of 39-43 amino acids derived from a larger beta APP. The proteolytic cleavage by 'alpha-secretase' generate soluble derivatives of beta APP (sAPP), lacking the cytoplasmic tail, transmembrane domain, and a small portion of the extracellular domain. Here levels of sAPP and beta APP were analyzed in cell lines of different origins by Western immunoblot of samples from conditioned media and cell lysates, respectively. Normal levels of secretion of sAPP into conditioned media were severely inhibited by treating different cell lines with a high dose of melatonin. In PC12 cells, levels of the fully matured beta APP forms of the post-Golgi compartment were more drastically decreased than the unglycosylated beta APP of the endoplasmic-reticulum (ER) forms. In other cell types, the unglycosylated ER-bound beta APP derivatives are predominant forms that were marginally affected by melatonin treatment. When the treatment of cells with melatonin was withdrawn, the normal level of secretion of sAPP was restored. Melatonin reduces the secretion of soluble A beta. Melatonin also inhibits the secretion of synaptophysin in PC12 cells. Taken together, these data suggest that melatonin probably affects the secretion of sAPP in the conditioned medium by interfering with its full maturation, and melatonin also affects the presysnaptic terminal marker.

Journal ArticleDOI
TL;DR: The results suggest that the protection afforded by the pineal hormone against indomethacin‐induced gastric injury may be, in addition to other possible mechanisms, to its radical scavenging activity.
Abstract: The gastric injury associated with nonsteroidal anti-inflammatory drug (NSAID) therapy has been linked to the detrimental effects of the agents on the processes of prostaglandin synthesis, neutrophil (PMN) activation, and oxygen free radical generation In the present study, we investigated the in vivo protective effects of melatonin on indomethacin-induced gastric lesions in the rat Peroxidation of lipids and changes in the activities of related enzymes such as glutathione peroxidase (GSH-px) and myeloperoxidase (MPO), as a marker of PMNs infiltration, were also studied Intraperitoneal (i p) injection of melatonin (025, 05, 1 mg kg−1) 30 min before indomethacin administration prevented gastric injury The mean ulcer indices significantly (P < 005) decreased Thiobarbituric acid (TBA) reactive substances in the gastric mucosa as an index of peroxidation, was increased after indomethacin administration and this increase was inhibited by melatonin In addition, pretreatment with melatonin resulted in a significant increase of the enzymatic GSH-px activity up to the control levels; however, inhibition of ulceration by melatonin was not associated with a significant reduction in PMN infiltration These results suggest that the protection afforded by the pineal hormone against indomethacin-induced gastric injury may be, in addition to other possible mechanisms, to its radical scavenging activity

Journal ArticleDOI
TL;DR: Melatonin circadian profile was not disrupted in 37 young male volunteers submitted to a typical pattern of exposure to the electromagnetic fields generated by two common types of cell phones.
Abstract: A decrease in melatonin secretion has been observed in small mammals under exposure to extremely low frequency electromagnetic fields. As there is some concern about possible health effects of the increasing use of radiocellular telephones emitting radiofrequency electromagnetic fields, we examined whether such fields would alter melatonin levels in the human. Volunteers were two groups totalling 38 men, 20–32 yr old. Exposures were to commercially available cellular telephones of the GSM 900 type (Global System for Mobile communication at 900 MHz) or DCS 1800 type (Digital Communication System at 1800 MHz), for 2 hr/day, 5 days/wk, for 4 wk, at their maximum power. Attention of the volunteers was sustained by TV projection of movies. Blood samples were collected hourly during the night and every 3 hr in the daytime. Four sampling sessions were performed at 15-day intervals: before the beginning of the exposure period, at the middle and the end of the exposure period, and 15 days later to evaluate the persistence or late appearance of potential effects. Evaluated parameters were the maximum serum concentration, the time of this maximum, and the area under the curve of the hormone profile. Melatonin circadian profile was not disrupted in 37 young male volunteers submitted to a typical pattern of exposure to the electromagnetic fields generated by two common types of cell phones.

Journal ArticleDOI
TL;DR: The preferential suppression of oxidative damage of proteins in Drosophila melanogaster strain Canton S indicates a particular protective role for melatonin in the aqueous phase of cellular compartments.
Abstract: In Drosophila melanogaster strain Canton S, the catalase inhibitor 3-amino-1,2,4-triazole was used to enhance oxidative stress from endogenous sources. This treatment was chosen as an alternative to direct administration of oxidants, which would cause damage and interact with melatonin already in the extracellular space before they reach the intracellular compartments. Male flies were kept in constant darkness and fed 1% sucrose as the only diet, with or without additions of melatonin (2 mM), inhibitor (100 mM), or combinations of both. After 20 or 24 hr, most of the animals exposed to 3-amino-1,2,4-triazole only had died, whereas a large number of flies had survived the inhibitor treatment in the presence of melatonin. Protein carbonyl, an indicator of oxidative protein modification, and lipid peroxidation, as determined by the formation of malondialdehyde and 4-hydroxyalkenal, were measured in flies treated for 20 hr. Melatonin alone did not substantially change these parameters, but prevented the increase in protein carbonyl caused by the catalase inhibitor. The effect of 3-amino-1,2,4-triazole on lipid peroxidation was relatively minor, but a clear-cut inhibition was found after simultaneous administration of melatonin. The preferential suppression of oxidative damage of proteins, as compared to lipids, indicates a particular protective role for melatonin in the aqueous phase of cellular compartments.

Journal ArticleDOI
TL;DR: The findings show that one pineal‐mediated effect on the immune system may be a direct action of melatonin onphagocytosis and the phagocytic biochemical process, and that this neurohormone might act as an antioxidant.
Abstract: A functional role for melatonin is its relationship to circadian timing mechanisms. In addition, there has recently been assumed to be a functional connection between the pineal gland and the immune system in mammals and birds, with some findings showing melatonin to be a free radical scavenger and general antioxidant. The present study investigates the possible relationship between the circadian rhythm of melatonin and the ingestion capacity as well as superoxide anion levels of ring dove (Streptopelia risoria) heterophils. In birds, heterophils, with their ability to ingest and kill different antigens, play a central role in the host defence mechanism. All determinations were made during 24 hr periods at 2 hr intervals. Radioimmunoassay showed an increase of melatonin serum levels during the dark period (from 20:00 to 07:00 hr) with a maximum at 04:00 hr, and a significant decline during the hours of light with a minimum at 16:00 hr. Similarly, the phagocytic index was enhanced during the night, with the maximum at approximately 04:00 hr and the minimum at approximately 18:00 hr. The same was the case in relation to phagocytic percentage. However, the superoxide anion levels were lower during darkness (minimum at 04:00 hr) and higher during the light period (maximum at 14:00 hr). In conclusion, our findings show that one pineal-mediated effect on the immune system may be a direct action of melatonin on phagocytosis and the phagocytic biochemical process, and that this neurohormone might act as an antioxidant.

Journal ArticleDOI
TL;DR: The results indicate that photoperiodic effects on T cell‐dependent humoral immunity are dependent upon the pineal gland, and raise the possibility that day length‐associated changes in some immune system functions are mediated by the Pineal melatonin rhythm.
Abstract: The present study tested the hypothesis that antibody production in response to xenoantigen is modulated by daylength and dependent upon the pineal gland. After injection of sheep erythrocytes (SRBC), serum immunoglobulin (Ig) concentrations were 5-fold lower in hamsters in short versus long days. Pinealectomy (Pinx) abolished the nocturnal melatonin rhythm, blocked short-day-mediated testis regression, and eliminated the short-day reduction in Ig production after SRBC treatment. Antibody titers in response to SRBC were equivalently augmented in short-day Pinx and long-day sham hamsters. The results indicate that photoperiodic effects on T cell-dependent humoral immunity are dependent upon the pineal gland. These findings raise the possibility that day length-associated changes in some immune system functions are mediated by the pineal melatonin rhythm.

Journal ArticleDOI
TL;DR: The data suggest that PKA type II plays an important role in the transcriptional control of melatonin biosynthesis in the rat pineal organ.
Abstract: Phosphorylation of cyclic AMP response element binding protein (CREB) at amino acid serine 133 appears as an important link between the norepinephrine (NE)-induced activation of second messenger systems and the stimulation of melatonin biosynthesis. Here we investigated in the rat pineal gland: 1) the type of protein kinase that mediates CREB phosphorylation; and 2) its impact on melatonin biosynthesis. Immunochemical or immunocytochemical demonstration of serine133-phosphorylated cyclic AMP regulated element binding protein (pCREB) and radioimmunological detection of melatonin revealed that only cyclic AMP-dependent protein kinase (PKA) inhibitors suppressed NE-induced CREB phosphorylation and stimulation of melatonin biosynthesis, whereas inhibitors of cyclic GMP-dependent protein kinase (PKG), mitogen-activated protein kinase kinase, protein kinase C, or calcium-calmodulin-dependent protein kinase (CaMK) were ineffective. Investigations with cyclic AMP-agonist pairs that selectively activate either PKA type I or II link NE-induced CREB phosphorylation and stimulation of melatonin biosynthesis to the activation of PKA type II. Our data suggest that PKA type II plays an important role in the transcriptional control of melatonin biosynthesis in the rat pineal organ.

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TL;DR: It is demonstrated that melatonin exerts potent anti‐inflammatory effects and part of these anti-inflammatory effects may be related to a reduction of prostaglandin production during the inflammatory process.
Abstract: The aim of the present study was to investigate the effect of melatonin on the production of the inflammatory mediator prostaglandins in a model of acute inflammation, carrageenan-induced pleurisy, where prostaglandins are known to play a crucial role. The results show that melatonin (12.5, 25, and 50 mg/kg, intraperitoneally, 15 min before the carrageenan) inhibits the inflammatory response (pleural exudate formation, polymorphonuclear cell infiltration, and prostaglandin production) in a dose-dependent manner. Immunohistochemical examination demonstrated a marked increase in the immunoreactivity to cyclooxygenase-2 (COX-2) in the lungs of carrageenan-treated rats. Pretreatment of carrageenan-treated rats with melatonin (50 mg/kg, intraperitoneally, 15 min before the carrageenan) prevented COX-2 expression. The present results demonstrate that melatonin exerts potent anti-inflammatory effects. Part of these anti-inflammatory effects may be related to a reduction of prostaglandin production during the inflammatory process.

Journal ArticleDOI
TL;DR: It was hypothesized that increased levels of this hormone in the gastrointestinal tract may play an important protective role against the development of colorectal cancer via stimulation of the immune system, protection against free radicals, and interaction with fatty acid uptake and metabolism.
Abstract: The distinct melatonin rhythm with higher concentrations during the darktime was found in plasma of both control patients and patients with colorectal carcinoma. Moderate surgery did not induce any changes in plasma melatonin levels, but a pronounced increase in both the day- and nighttime melatonin concentrations was found after surgical treatment for colon cancer. The melatonin content in the tumor tissue did not differ from that in the proximal and the distal parts of the resected gut, which were without signs of malignant changes. Neither concentrations of serotonin nor 5-hydroxyindole acetic acid differed among analyzed parts of the gut. Daytime melatonin concentrations in gut tissue (314.7 +/- 87.8 pg/g of wet tissue) were more than ten times higher than the daytime levels in circulation. It was hypothesized that increased levels of this hormone in the gastrointestinal tract may play an important protective role against the development of colorectal cancer via stimulation of the immune system, protection against free radicals, and interaction with fatty acid uptake and metabolism.

Journal ArticleDOI
TL;DR: It is suggested that the avian embryo has evolved its own early circadian melatonin‐producing system because, as a consequence of its extrauterine development, it cannot use the system of its mother.
Abstract: In contrast to the situation in mammals, in which circadian melatonin production by the pineal gland does not begin until some time after birth, the development of pineal gland rhythmicity is an embryonic event in the precocial domestic fowl. A distinct melatonin rhythm was found in 19-d-old chick embryos maintained under light:dark (LD) 16:8. No significant variation in melatonin levels was detected in embryos exposed to LD 8:16. The melatonin rhythm in the pineal gland and plasma of chick embryos incubated for 18 d in LD 12:12 persisted for 2 d in constant darkness indicating that melatonin production is under circadian control at least from the end of embryonic life. A 1-d exposure to a LD cycle during the first postembryonic day was sufficient to entrain the melatonin rhythm, and previous embryonic exposure to either LD or constant darkness (DD) neither modified this rapid synchronization nor did it affect the melatonin pattern during the two subsequent days in DD. It is suggested that, in contrast to the situation in mammals, the avian embryo has evolved its own early circadian melatonin-producing system because, as a consequence of its extrauterine development, it cannot use the system of its mother.

Journal ArticleDOI
TL;DR: Light‐mediated events on the parasite biological cycle could be mediated by light‐induced changes in melatonin produced by this unicell, and the participation of the indoleamine in its biological cycle is suggested.
Abstract: The unicellular organism Trypanosoma cruzi is an eukaryote whose cell cycle mainly occurs under darkness in the insect gut. The unique external phase corresponds to the metacyclic forms, the forms that are able to infect humans, which appear within the insect deyections. Thus, light may be a powerful stressor in this unicell. Epimastigote forms (the parasite forms that grow and transform to metacyclic forms in the insect gut) of Trypanosoma cruzi grow normally when cultured in a LD cycle of 0:24 hr, reaching exponential growth by the 7th day. A pulse of 2 hr of light (LD 2:22) was enough to block the growth of the epimastigotes, an effect that was correlated with the expression of heat-shock proteins during the first 120 min of light exposure. Thereafter, protein synthesis decreased. Light exposure of metacyclic forms also inhibits the parasitization ability. It is known that light regulates the production of melatonin in most animal species studied, including other unicells such as dinoflagellates. T. cruzi contains and synthesizes melatonin and, thus, light-mediated events on the parasite biological cycle could be mediated by light-induced changes in melatonin produced by this unicell. Epimastigotes cultured under continuous darkness produce melatonin over the 24 hr period in a biphasic manner. Coinciding with the melatonin peaks, there was high melatonin efflux from the parasite into the medium. Epimastigotes cultured for 7 days under a LD cycle of 2:22 hr showed a 55% reduction in melatonin content, although this reduction seems not to be related with the growth delay. In fact, incubation of epimastigotes with exogenous melatonin (1 pM) did not affect parasite growth, but significantly reduced their transformation into metacyclic forms by the 7-8th day of treatment. Thus, the light-dependent decrease in melatonin production by the unicell may be responsible, at least partially, for the light-induce parasitization inhibition. Moreover, melatonin production is highest in the metacyclic forms. These data support a link between light, melatonin production and parasitization ability of T. cruzi and suggest the participation of the indoleamine in its biological cycle.

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TL;DR: Melatonin may contribute to the realization of chemotherapy in metastatic cancer patients unable to tolerate the chemotherapeutic approach because of persistent thrombocytopenia, and to prevent chemotherapy‐induced platelet decline.
Abstract: Thrombocytopenia is a frequent complication of cancer and constitutes an absolute contraindication for chemotherapy. Recent studies have demonstrated that platelet generation may be influenced by both cytokines and neurohormones. In particular, the pineal indole melatonin has been proven to enhance platelet number in patients with thrombocytopenia due to different reasons. On this basis, we have evaluated the effects of a concomitant administration of melatonin in thrombocytopenic cancer patients undergoing chemotherapy. The study was performed in 14 metastatic breast cancer women treated by weekly epirubicin. Each cycle consisted of epirubicin at 25 mg/m2 i. v. at weekly intervals. Melatonin was given orally at 20 mg/day in the evening every day, starting 7 days prior to chemotherapy. Patients were considered as evaluable when they received at least four cycles of chemotherapy. Evaluable patients were 12/14. The induction phase with melatonin induced a normalization of platelet number in 9/12 evaluable patients, and no further platelet decline occurred on chemotherapy. Objective tumor regression was achieved in 5/12 (41%) patients. This preliminary study would suggest that melatonin may be effective in the treatment of cancer-related thrombocytopenia and to prevent chemotherapy-induced platelet decline. Until now, melatonin therapy of cancer has been generally considered as an alternative treatment to chemotherapy. In contrast, this study would suggest that melatonin may contribute to the realization of chemotherapy in metastatic cancer patients unable to tolerate the chemotherapeutic approach because of persistent thrombocytopenia.

Journal ArticleDOI
TL;DR: The first demonstration of direct interaction of melatonin with hormone‐insensitive prostate tumor cells is provided, coupled to pertussis toxin‐sensitive G proteins to induce cell density‐dependent changes in cGMP, cAMP, and cell growth.
Abstract: Melatonin, secreted nocturnally by the pineal gland, can bind to human benign prostate epithelial cells and attenuate their growth and viability. In the present study, melatonin binding and responses were explored in the human steroid-independent PC3 prostatic tumor cells. PC3 cells bound 125I-melatonin with low affinity (Kd ca. 0.9 nM) at high as well as low cell density. Melatonin enhanced cGMP and 3H-thymidine incorporation at low, but attenuated them at high cell density. In addition, melatonin inhibited cAMP at low, but augmented it at high cell density. These effects were associated with an increase in cell count at low- but not high-density cultures. Pertussis toxin treatment suppressed 125I-melatonin binding and ablated all the effects of melatonin on 3H-thymidine incorporation, cAMP, and cGMP at both cell densities. Cholera toxin treatment failed to block the effects of melatonin on 3H-thymidine incorporation, but prevented the modulation by melatonin of cAMP at low and cGMP at high cell density. The cGMP analog 8-Br-cGMP, inhibited melatonin's effects on 3H-thymidine incorporation at both cell densities. H89, a protein kinase A inhibitor, prevented melatonin's effects on 3H-thymidine incorporation at low but not high cell density. These results provide the first demonstration of direct interaction of melatonin with hormone-insensitive prostate tumor cells. The melatonin receptors in the PC3 cells are coupled to pertussis toxin-sensitive G proteins to induce cell density-dependent changes in cGMP, cAMP, and cell growth.

Journal ArticleDOI
TL;DR: It is concluded that CR3 receptors, MHC antigens, and CD4 antIGens on macrophages/microglia are upregulated following melatonin administration.
Abstract: The present study examined the response of macrophages/microglia to multiple injections of melatonin in the pineal gland and different regions of the brain. The macrophages/microglia showed a significant increase in cell numbers and upregulation of complement type 3 receptors (CR3), major histocompatibility complex class I (MHC I) and class II (MHC II) antigens, and antigens of monocyte/macrophage lineage, as detected by the antibodies OX-42, OX-18, OX-6, and ED1, respectively. The upregulation of the above antigens was observed in 1-d-old rats given daily injections of melatonin and killed at 7-11 d of age; no noticeable change was observed at earlier time intervals. The macrophages/microglia expressing the above antigens appeared round and showed a vacuolated cytoplasm compared with ramified cells in the control rats. Upregulation of CD4 antigens as detected with the antibody W3/25 was also observed in macrophages/microglia in the corpus callosum and epiplexus cells in the lateral ventricles, but not in the pineal gland and the cerebral cortex in the same age group. In rats killed between 2 and 5 d, and at 14 d of age after melatonin treatment, the immunoreactivities of macrophages/microglia with the above mentioned antibodies were comparable to cells in the control rats. Immunoreactive cells were not detected in any of the age groups in melatonin-treated or control rats with the antibodies W3/13 and OX-33, which are markers for T and B lymphocytes. It is concluded that CR3 receptors, MHC antigens, and CD4 antigens on macrophages/microglia are upregulated following melatonin administration. On the other hand, once the melatonin treatment is discontinued the expression of the various antigens/receptors returns to normal levels, suggesting that increased immune potentiality and its maintenance in these cells require the continuous action of the drug.

Journal ArticleDOI
TL;DR: Melatonin may have a beneficial effect by suppressing the expression of TF activity in LPS‐stimulated monocytes of whole blood, which may be physiologically important in upregulation of the immune system.
Abstract: The pineal hormone melatonin, due to its lipophilic nature, has access to every cell and every part of a cell in the body, suggesting that it could exert effects on blood immune cells. The regulation of the activation of monocytes may be important in a number of diseases, especially pathophysiological conditions associated with inflammatory reactions. Considering this, a study on the effect of melatonin on monocytes in whole blood was carried out. Melatonin added at a final concentration of 5 ng/mL to whole blood in vitro reduced lipopolysaccharide (LPS)-induced tissue factor (TF) activity in monocytes by 55% in blood from a group of subjects with melatonin-sensitive cells. At even lower concentrations of melatonin (20-50 pg/mL) and in the physiological range, a trend of suppressed LPS-induced TF activity by approximately 20% was seen. A further indication of a downregulation of LPS-stimulated monocytes by melatonin was shown by its reduction of LPS-induced tumor necrosis factor (TNF). Twenty to one hundred pg/mL melatonin caused a significant reduction of LPS-induced TNF production by approximately 25-30%. In contrast, melatonin at a final concentration of 10 pg/mL, added to whole blood incubated with LPS and also the phorbol ester, PMA, caused a significant rise of 25%; whereas 100 pg/mL enhanced LPS + PMA-induced TNF by approximately 80% as compared to LPS + PMA alone. These effects were not detectable during the winter darkness of Tromso (70 degrees N), probably due to the high content of melatonin in the blood even at daytime. These results show that melatonin may have a beneficial effect by suppressing the expression of TF activity in LPS-stimulated monocytes. Furthermore, the results indicate that LPS-induced TF in monocytes of whole blood is independent of protein kinase C (PKC) activation. Melatonin is probably amplifying cellular activation reactions that are PKC-dependent. This may be physiologically important in upregulation of the immune system.