A glassy carbon electrode modified with a composite consisting of electrodeposited chitosan and carboxylated multi-walled carbon nanotubes (e-CS/MWCNTs/GCE) was used as a working electrode for simultaneous determination of dopamine (DA), serotonin (5-HT) and melatonin (MT), which were related to circadian rhythms. The electrochemical characterizations of the working electrode were carried out via electrochemical impedance spectroscopy and chronocoulometry. It was found that electrochemical modification method, that was cyclic voltammetry, may can cause continuous CS polymerization on MWCNTs surface to form a dense membrane with more active sites on the electrode, and the electrochemically active surface area of e-CS/MWCNTs/GCE obtained was about 7 times that of GCE. The electrochemical behaviour of DA, 5-HT and MT on working electrode were carried out via differential pulse voltammetry and cyclic voltammetry. The results showed that e-CS/MWCNTs/GCE solved the problem that the bare electrode could not detect three substances simultaneously, and can catalyze oxidation potential difference as low as 0.17 V of two substances reaction at the same time, indicating very good electrocatalytic activity. By optimizing the detection conditions, the sensor showed a good linear response to DA, 5-HT and MT in the range of 20-1000 μmol/L, 9-1000 μmol/L and 20-1000 μmol/L, and the detection limits were 12 μmol/L, 10 μmol/L and 22 μmol/L (S/N = 3), respectively. In addition, the proposed sensor was successfully applied to the simultaneous detection of DA, 5-HT and MT in human saliva samples.

어류의 번식은 뇌에서 분비되는 다양한 신경호르몬과 뇌하수체에서 분비되는 생식소 자극 호 르몬에 의해 조절된다. 극동산 뱀장어(Anguilla japonica)의 번식도 이 호르몬들의 작용에 의해 조절되지만 성 성숙 시 신경호르몬이 뇌하수체 호르몬을 조절하는 방법은 완전히 밝혀지지 않 았다. 이전 연구에 의하면 progesterone (P4), melatonin 및 serotonin (5-HT) 등과 같은 신경호 르몬이 일부 어류의 번식 과정 조절에 관여하는 것으로 밝혀졌다. 본 연구에서는 뱀장어의 뇌 하수체를 초대 배양하였고, 안정화된 뇌하수체 세포에 P4, 17β-estradiol (E2), melatonin 및 5- HT를 처리하였다. 이후 처리된 호르몬의 작용이 뇌하수체 세포에서 번식 관련 호르몬인 FSHβ, LHβ, GH 및 SL mRNA 발현에 어떤 영향을 미치는지 조사하였다. 본 연구를 수행한 결과, P4는 뇌하수체 세포에서 FSHβ와 LHβ 발현을 증가시켰고, melatonin은 FSHβ와 LHβ 뿐만 아니라 GH 와 SL의 발현을 증가시켰다. 하지만 5-HT는 이 유전자의 mRNA 발현에 유의한 영향을 미치지 않았다. 이상의 결과는 P4 또는 melatonin이 뱀장어의 초기 성 성숙에 중요한 역할을 할 수 있음을 의미한다.

견과류는 영양가가 우수하고 다양한 기능성 생리활성을 가져 건강한 식단에 중요한 요소이다. 멜라토닌은 뇌의 송 과선에서 분비되는 호르몬으로 각종 조직의 산화적 손상을 방지하는 것으로 알려져 있으며, 식물체에서도 중요한 역할 을 한다. 본 연구는 견과류의 멜라토닌 추출 및 정량을 위 한 분석법을 마련하였고, 견과류의 높은 지방함량을 고려한 최적화된 추출 절차를 이용하여 경기도에서 유통되는 견과 종실류 등의 멜라토닌 함량을 조사하였다. 땅콩 또는 견과 류, 유지종실류 및 두류의 평균 멜라토닌 함량은 각각 1200.83 (409.76-2223.56), 934.83 (454.10-1736.60), 616.46 (494.70- 825.12) pg/g으로 나타났다. 호두와 밤의 경우 속껍질을 포 함한 반탈각 상태와 과실만 있는 경우의 멜라토닌 함량을 비교한 결과, 반탈각 상태에서 멜라토닌 함량이 유의적으로 높았다. 또한 호두, 밤 및 땅콩의 경우 2022년 재배된 햇 견 과보다 2021년 재배 후 저장된 견과에서 멜라토닌 함량이 유의적으로 높았다. 이러한 결과는 견과종실류 등의 멜라토 닌 함량이 높아 우수한 기능성 식품임을 시사하였다.

Lipopolysaccharide (LPS) is an endotoxin factor present in the cell wall of Gram-negative bacteria and induces various immune responses to infection. Recent studies have reported that LPS induces cellular stress in various cells including oocytes and embryos. Melatonin (N-acetyl-5-methoxytryptamine) is a regulatory hormone of circadian rhythm and a powerful antioxidant. It has been known that melatonin has an effective function in scavenging oxygen free radicals and has been used as an antioxidant to reduce the cytotoxic effects induced by LPS. However, the effect of melatonin on LPS treated early embryonic development has not yet been confirmed. In this study, we cultured mouse embryos in medium supplemented with LPS or/and melatonin up to the blastocyst stage in vitro and then evaluated the developmental rate. As a result of the LPS-treatment, the rate of blastocyst development was significantly reduced compared to the control group in all the LPS groups. Next, in the melatonin only treated group, there was no statistical difference in embryonic development and no toxic effects were observed. And then we found that the treatment of melatonin improved the rates of compaction and blastocyst development of LPS-treated embryos. In addition, we showed that melatonin treatment decreased ROS levels compared to the LPS only treated group. In conclusion, we demonstrated the protective effect of melatonin on the embryonic developmental rate reduced by LPS. These results suggest a direction to improve reproduction loss that may occur due to LPS exposure and bacterial infection through the using of melatonin during in vitro culture.

Melatonin is a neurotransmitter that modulates various physiological phenomena including regulation and maintenance of the circadian rhythm. Nicotinic acetylcholine receptors (nAChRs) play an important role in oral functions including orofacial muscle contraction, salivary secretion, and tooth development. However, knowledge regarding physiological crosstalk between melatonin and nAChRs is limited. In the present study, the melatoninmediated modulation of nAChR functions using bovine adrenal chromaffin cells, a representative model for the study of nAChRs, was investigated. Melatonin inhibited the nicotinic agonist dimethylphenylpiperazinium (DMPP) iodide-induced cytosolic free Ca2+ concentration ([Ca2+]i) increase and norepinephrine secretion in a concentrationdependent manner. The inhibitory effect of melatonin on the DMPP-induced [Ca2+]i increase was observed when the melatonin treatment was performed simultaneously with DMPP. The results indicate that melatonin inhibits nAChR functions in both peripheral and central nervous systems.

Melatonin (N-aceyl-5-methoxytryptamine) is the major hormone of the pineal gland. Melatonin and its metabolic derivatives possess extensive free-radical scavenging abilities and played critical roles in antioxidative stress, resisting apoptotic cell death. Melatonin also could enhance mitochondrial biogenesis in rats with carbon tetrachloride-induced liver fibrosis. In addition, melatonin attenuates myocardial ischemia/reperfusion injury by reducing oxidative stress damage via activation of SIRT1 signaling in a melatonin receptor 2-dependent manner. Activation or overexpression of SIRT1 could enhance mitochondrial biogenesis and function by inducing PGC-1α expression and deacetylation. The aim of this study was to investigate if melatonin enhances mitochondrial biogenesis and function via activation of melatonin receptor 2/SIRT1/PGC1-α Pathway. The results showed that Melatonin rescued rotenone-induced impairment of porcine embryo development. Treatment with rotenone could increase oxidative stress and apoptosis. Rotenone impaired mitochondrial functions by disrupting mitochondrial membrane potential, reducing mitochondrial DNA copy number and ATP production. Melatonin could improve SIRT1 and PGC-1α expression, inducing mitochondrial biogenesis. Rotenone-induced mitochondrial dysfunction and ATP deficiency was rescued by melatonin treatment, the oxidative stress and apoptosis was significantly decreased. Inhibition of melatonin receptor 2 or Knockdown of SIRT1 abolished the protective effects of melatonin on rotenone-induced impairments. Therefore, melatonin enhanced mitochondrial biogenesis and function, protected against rotenone-induced impairments.

In the present study, we investigated the physiological effects of smartphone use at night when the display luminance and white balance were differently manipulated. Two levels of luminance and two types of white balance were combined to form four types of displays. Subjects were instructed to use smartphones between 23:00 to 01:00 twice a week for two weeks, and for each trial, subjects were given one of the four display types. Melatonin concentration in the saliva, body temperature and heart rate were measured before and after each experiment. The experimental result showed that the low luminance display supported melatonin secretion and thermoregulation compared to the high luminance display. With regard to the white balance, higher melatonin level was observed when using the display that filtered blue light. The low luminance display together with yellowish tint best supported restful sleep at night in terms of every physiological response. This study collectively demonstrates that bright and blue light emitted from smartphone displays adversely affect melatonin secretion, body temperature, and heart rate, and therefore, suggests the use of a display with low luminance or a display that filters blue light for a restful sleep at night.

Although anti-aging activities of melatonin, a hormone secreted by the pineal gland, have been reported in senescence-accelerated mouse models and several types of cells, its impact and mechanism on the senescence of human dental pulp cells (HDPCs) remains unknown. In this study, we examined the impact of melatonin on cellular premature senescence of HDPCs. Here, we found that melatonin markedly inhibited senescent characteristics of HDPCs after exposure to hydrogen peroxide (H2O2), including the increase in senescence-associated β-galactosidase (SA-β-gal)-positive HDPCs and the upregulation of p21 protein, an indicator for senescence. In addition, as melatonin attenuated H2O2-stimulated phosphorylation of c-Jun N-terminal kinase (JNK), while selective inhibition of JNK activity with SP600125 significantly attenuated H2O2-induced increase in SA-beta-gal activity. Results reveal that melatonin antagonizes premature senescence of HDPCs via JNK pathway. Thus, melatonin may have therapeutic potential to prevent stress-induced premature senescence, possibly correlated with development of dental pulp diseases, and to maintain oral health across the life span.

Melatonin has an important role as anti-oxidative effect and reducing of endoplasmic reticulum(ER)-stress on oocyte maturation and embryo development. Under ER-stress condition, unfolding protein response (UPR) is a defence mechanism in mammalian cells. Recently, regulation of UPR signaling genes are involved in oocyte maturation, embryo development and female reproduction. However, there is no report on the role of melatonin for UPR signaling and ER-stress mediated apoptosis during pig oocyte maturation progression. Moreover, the changes of UPR genes expression according to the porcine oocyte maturation is not yet fully understood. Here, we investigated the changes of UPR signal (BIP/GRP78, ATF4, p90/p50ATF6, and XBP1) and ER-stress apoptotic factor CHOP genes expressions in porcine oocyte maturation by Western blot and RT-PCR analysis. During oocyte maturation, UPR marker and CHOP genes expressions were significantly increased in matured oocytes or cumulus-oocyte complexes (COCs). UPR markers expressions were significantly increased by ER-stress inducer, tunicamycin (Tm), treated (1, 5, 10 μg/ml) groups in a dose-dependent manner compared with control group. To confirm the reducing of ER-stress by melatonin (0.1 μM), we were compared to the effects of ER-stress inhibitor, TUDCA (200 μM), after pre-treated Tm (5 μg/ml) for 22 h maturation. Expressions of UPR markers and meiotic maturation were recovered by melatonin (0.1 μM) in COCs. And, we observed the role of Grp78/Bip as UPR signaling beginning marker using siRNA. In result, reduction of Grp78/Bip gene expression by siRNA was induced the inhibition of oocyte maturation (32.5±10.1 vs control; 77.8±5.3), and p50ATF6 protein level was significantly decreased (p<0.001) in cultured COCs for 44 h. In addition, these results were recovered through the addition of melatonin (0.1 μM) or TUDCA (200 μM) in maturation medium. These results demonstrated that the regulation of UPR signaling via Grp78/Bip gene induction plays a critical role in porcine oocyte maturation in vitro. Furthermore, this present study first confirmed a functional link between inhibition effect of ER-stress by melatonin and regulating of UPR signaling in porcine oocyte maturation. In conclusion, melatonin improves the oocyte maturation and cumulus cells expansion of COCs through the regulation of UPR signal pathway by BIP/GRP78 against the ER-stress during porcine oocyte maturation periods.

In this study, to improve the in vitro development of various cells including cloned embryos, the effects that isoproterenol and melatonin have on in vitro development of porcine parthenogenetic oocytes were investigated. Parthenogenetic activation was induced with electrical stimulation, BSA and 6-DMAP treatment. 10-7 M of melatonin and isoproterenol (10-10, 10-12 and 10-14 M) were supplemented for in vitro maturation (IVM) and in vitro culture (IVC) medium, with different concentrations. When isoproterenol and melatonin were supplemented in IVM medium with different concentrations, there was no significant (P<0.05) difference of maturation rate in the treatment groups as well as in that of only melatonin. As isoproterenol and melatonin were supplemented in IVM medium with different concentrations, blastocyst rates of isoproterenol 10-12 M treatment group (37.1%) were significantly (P<0.05) higher than control group (26.0%). Isoproterenol and melatonin were supplemented in IVC medium with different concentrations, then the cleavage rate of 10-12 M isoproterenol treatment group (82.2%) was significantly (P<0.05) higher than the group that melatonin was only supplemented (70.9%). There was no difference of blastocyst rate between the treatment groups. When isoproterenol and melatonin were supplemented for IVM+IVC medium with different concentrations, the cleavage rate of 10-12 M isoproterenol treatment group (92.5%) was significantly (P<0.05) higher than the control group (82.8%) and the group that melatonin was only treated (81.6%). The blastocyst rate of 10-12 M as 45.6% was significantly (P<0.05) higher than control group (25.2%) and melatonin treatment group (31.2%). The cell number of blastocyst in 10-12 M isoproterenol treatment group 35.5±3.4 was significantly (P<0.05) highest. The results of this study showed that the development rate of IVC when both isoproterenol and melatonin were supplemented was higher than when melatonin was only supplemented. Therefore, it is concluded that isoproterenol is rather effective in the activation of melatonin. 10-7 M melatonin and 10-12 M isoproterenol were considered suitable concentration.

Acute ischemic stroke results from sudden decrease or loss of blood supply to an area of the brain, resulting in a coinciding loss of neurological function. The antioxidant action of melatonin is an important mechanism among its known effects to protective activity during ischemic/reperfusion injury. The focus of this research, therapeutic efficacy of melatonin on recovery of neurological function following long term treatment in ischemic brain injured rats. Male Sprague-Dawley rats (n=40; 8 weeks old) were divided into the control group, and MCAo groups (Vehicle, MT7 : MCAo+ melatonin injection at 7:00, MT19 : MCAo+melatonin injection at 19:00, and MT7,19 : MCAo+melatonin injection at 7:00 and 19:00). Rat body weight and neurological function were measured every week for 8 weeks. After 8 weeks, the rats were anesthetized with a mixture of zoletil (40 mg/kg) and xylazine (10 mg/kg) and sacrificed for further analysis. Tissues were then collected for RNA isolation from brain tissue. Also, brain tissues were analyzed by histological procedures. We elucidated that melatonin was not toxic in vital organs. MT7,19 was the most rapidly got back to mild symptom on test of neurological parameter. Also, exogenous melatonin induces both the down-regulation of detrimental genes, such as NOSs and the up-regulation of beneficial gene, including BDNF during long term administration after focal cerebral ischemia. Melatonin treatment reduced the loss of primary motor cortex. Therefore, we suggest that melatonin could be act as prophylactic as well as therapeutic agent for neurorehabilitative intervention.

Melatonin (N-acetyl-5-methoxytryptamine) is the major neurohormone secreted during the night by the vertebrate pineal gland. The circadian pattern of pineal melatonin secretion is related to the biological clock within the suprachiasmatic nucleus (SCN) of the hypothalamus in mammals. The SCN coordinates the body's rhythms to the environmental light-dark cycle in response to light perceived by the retina, which acts mainly on retinal ganglion cells that contain the photopigment melanopsin. Calbindin-D9k (CaBP-9k) is a member of the S100 family of intracellular calcium- binding proteins, and in this review, we discuss the involvement of melatonin and CaBP-9k with respect to calcium homeostasis and apoptotic cell death. In future studies, we hope to provide important information on the roles played by CaBP-9k in cell signal transduction, cell proliferation, and homeostasis in vivo and in vitro.