Day time-restricted feeding shows differential synchronizing effects on age-related changes of serotonin metabolism in SCN and the pineal gland in male Wistar rats.

The circadian timing system is synchronized by the environmental photic and non-photic signals. Light is the major cue that entrains the master circadian oscillator located in suprachiasmatic nucleus (SCN). With aging condition ocular light impairs because of the age-related deficiencies in the eye as a result the clock becomes less sensitive to light. In such case non-photic cues may play a major role in synchronizing the clock. Earlier studies have linked altered meal timings to induce many physiological changes including serotonin in different brain regions such as hypothalamus, brain stem and striatum. Much is not known about the effect of timed food restriction as a non-photic stimulus on serotonergic system in SCN under aging condition. We report here the synchronizing effects of time-restricted feeding (TRF) as a non-photic stimulus on serotonin and its related metabolites in the SCN and pineal gland of male Wistar rats upon aging. Under food restriction daily rhythmicity of serotonin 5-HT and 5-HTOH was abolished whereas NAS, 5-MIAA and NAT showed a significant decrease in their daily pulses upon food restriction in 3 months (m) old rats. Under forced day time feeding schedule the mean 24 h levels of serotonin have significantly decreased in 12 and 24 m old animals in SCN and pineal gland. Most of the serotonin metabolites in the SCN and pineal gland of 12 and 24 m old ad libitum fed group rats have shown rhythmicity. 5-HT, NAS, MEL and NAT have shown daily rhythm in the SCN of 12 and 24 m old rats whereas 5-MIAA and 5-MTOH did not show daily rhythm in both the age groups. The mean 24 h levels of 5-HTP, 5-HIAA, 5-MIAA, 5-MTOH, MEL and NAT were increased in the pineal gland of 12 and 24 months old rats. This work help demonstrate the role of TRF in synchronising age induced desynchronization in serotonin metabolome.

Therapeutic effects of hydro-alcoholic leaf extract of Withania somnifera on age-induced changes in daily rhythms of Sirt1, Nrf2 and Rev-erbα in the SCN of male Wistar rats.

The temporal expression pattern of the circadian clock genes are known to be altered/attenuated with advance in age. Withania somnifera (WS) essentially consists of numerous active constituents including withanolides is known to have antioxidant, anti-inflammatory and adaptogenic properties. We have earlier demonstrated therapeutic effects of hydro-alcoholic leaf extract of WS on the age-induced alterations in the levels and daily rhythms of various clock genes such as rBmal1, rPer1, rPer2 and rCry1. We have now studied effects of hydro-alcoholic leaf extract of WS on the age-induced alterations in the levels and daily rhythms of expression of SIRT1 (an NAD+ dependent histone deacetylase and a modulator of clock) and NRF2 (a clock controlled gene and a master transcription factor regulating various endogenous antioxidant enzymes) in addition to rRev-erbα in SCN of adult [3 months (m)], middle-aged (12 m) and old-aged (24 m) male Wistar rats. The daily rhythms of rNrf2 expression showed 6 h phase delay in middle age and 12 h phase advance in old age. WS restored rSirt1 daily rhythms and phase in old age whereas it restored the phase of rNrf2 in the SCN of both middle and old aged animals. At protein level, SIRT1 expression showed phase advances in 12 m and 24 m whereas NRF2 daily rhythms were abolished in both the age groups. WS restored the phase and daily rhythms of SIRT1 as well as NRF2 in 12 m old rats. However, rRev-erbα expression was found insensitive to WS treatment in all the age groups studied. Pairwise correlation analysis demonstrated significant stoichiometric interactions among rSirt1, rNrf2 and rRev-erbα in 3 m which altered with aging significantly. WS treatment resulted in differential restorations of such interactions.

Aging renders desynchronization between clock and immune genes in male Wistar rat kidney: chronobiotic role of curcumin.

Suprachiasmatic nucleus (SCN) contains the central clock that orchestrate circadian rhythms in physiology and behavior in mammals. Tightly interlocked transcriptional and translational feedback loops (TTFLs) comprising of various clock genes such as Clock, Bmal1, Periods, Cryptochromes etc. in the SCN, send the timing signals to peripheral clocks that governs local metabolism with similar TTFLs. Peripheral clocks in kidney regulates several circadian rhythms like blood pressure, immunity etc. However, aging leads to circadian and inflammatory disorders in kidney. Though there are increasing evidences on age associated perturbations, studies elucidating the rhythmic expression of clock and immune genes across aging in kidney are obscure. We therefore studied changes in daily rhythms of clock and immune genes in kidney. In this study we measured mRNA expression of clock genes rBmal1, rPer1, rPer2, rCry1, rCry2, rRev-erbα, rRorα, and inflammatory genes rNfκb1, rTnfα, rIl6, rTlr4 and rTlr9 in 3, 12 and 24 months male Wistar rat kidney using qRT-PCR. From our study, we did not observe significant changes in clock genes expression except rRorα, but immune genes showed significant phase alterations as well as increase in mean 24 h levels. Pearson correlation analysis of data showed desynchronization between immune and clock genes expression. We further studied the effect of administration of curcumin which has anti-aging, anti-inflammatory, anti-oxidant etc. properties, and evaluated its chronobiotic properties. We here report differential effects of curcumin administration on daily rhythms of clock and immune genes expression.

Therapeutic effects of curcumin on age-induced alterations in daily rhythms of clock genes and Sirt1 expression in the SCN of male Wistar rats.

The aging brain is linked to accumulation of oxidative stress and increase in damage to biomolecules which in turn may cause or promote circadian dysfunction by disruption of biological clock, the suprachiasmatic nucleus (SCN). Age associated alterations in clock gene expression in the SCN has been reported earlier. In the present study we have examined therapeutic effects of the antioxidant curcumin on age induced alterations in daily rhythms and levels of core clock genes in SCN of young [3 months (m)], middle (12 months) and old (24 months) male Wistar rats. Curcumin was administered orally at ZT-11, 1 hour (h) before the onset of darkness. The effect of curcumin administration on daily rhythms and levels of expression of clock genes such as rBmal1, rPer1, rPer2, rCry1, rCry2 and rRev-erbα as well as on the clock modulator rSirt1 were studied. There was restoration of phase of rPer1, rPer2, rCry1, rCry2 and daily pulse of rPer2 in middle aged animals. However, in old aged rats the phase and daily pulse of rPer1 were restored with curcumin treatment. rSirt1 did not show age related alterations in its transcript levels though the rhythms were abolished in old aged rat SCN. Pearson correlation analysis showed that curcumin administration to 12 and 24 months animals had resulted in restorations of several correlations among clock genes which were found to be altered/abolished in age matched control groups. In addition, strong interlocking interactions between rSirt1 and clock genes were observed in young age which were disrupted with aging and curcumin administration resulted in partial restoration.

Daily chronomics of proteomic profile in aging and rotenone-induced Parkinson's disease model in male Wistar rat and its modulation by melatonin.

Aging is associated with changes in several basic parameters of circadian timing system (CTS) in mammals leading to circadian dysfunction. We had reported earlier that upon aging and in rotenone induced Parkinson's disease (RIPD) rat model there were significant alterations in the core clock genes expression levels and daily pulses. To identify biomarkers of aging and PD chronomics of proteomic day-night profiles in suprachiasmatic nucleus (SCN), pineal and substantia nigra (SN) in 3 month (m), 12, 24 m and RIPD rat model were studied at two time points i.e. Zeitgeber Time (ZT)-6 (mid-day) and ZT-18 (mid-night). Proteome analysis was done by using two dimensional (2-D) electrophoresis and the spots showing robust day-night variations were identified by using MALDI TOF/TOF analysis. In 3 m rats the number of proteins showing day-night variations were relatively more than 12, 24 m and RIPD rat model in SCN and SN. But in pineal there was increase in number of protein spots showing day-night variations in 24 m. Mass spectroscopy of the protein spots showing robust day night variation in aging and RIPD rats were identified. As melatonin, a multitasking molecule, an endogenous synchronizer of rhythm, an antioxidant and an antiaging drug, declines with aging, the effects of melatonin administration on differential alterations in chronomics of 2-D protein profiles in aging and RIPD male Wistar rats were studied. We report here that the melatonin could be playing an important role in modulating the chronomics of 2-D protein profiles. Additionally, various proteins were identified for the first time in this study showing significant day night variation in SCN, pineal and SN may prove useful towards targeting novel treatments for circadian dysfunction, good health and longevity.

Daily Socs1 rhythms alter with aging differentially in peripheral clocks in male Wistar rats: therapeutic effects of melatonin.

Suprachiasmatic nucleus (SCN) in synchronization with the peripheral clocks regulates the temporal oscillations leading to overt rhythms. Aging leads to attenuation of such circadian regulation, accompanied by increased inflammatory mediators prevalently the cytokines. Suppressors of cytokine signaling (SOCS) family of proteins such as SOCS 1, 3 and cytokine-inducible SH2-containing protein (CIS) negatively regulate the cytokine signaling pathway. The role of SOCS1 in aging and circadian system is obscure. We therefore studied the daily rhythms of rSocs1 mRNA expression at Zeitgeber time (ZT) -0, 6, 12 and 18 in peripheral clocks such as liver, kidney, intestine and heart of 3, 12 and 24 months (m) old male Wistar rats. Interestingly the peripheral clocks studied displayed a rhythmic rSocs1 gene expression in 3 months. In 12 months group, 12 h phase advance in liver and 12 h phase delay in kidney and heart was observed with abolition of rhythms in intestine. Aging (24 months group) resulted in a phase advance by 6 h in liver and heart with abolition of rhythms in intestine in 24 months group. Kidney was also significantly affected upon aging with significant decrease in the rSocs1 levels and abolition of rhythms. The decrease in melatonin levels with aging is associated with decreased immunity and increased oxidative stress. The exogenous administration of melatonin has been linked to play a role in re-synchronization of circadian rhythms, reducing oxidative stress and enhancing immune properties. We therefore had studied the effect of exogenous melatonin upon age induced changes in daily rSocs1 gene expression patterns. Melatonin treatment partially restored the rhythms and daily pulse (ratio of maximum:minimum levels) in liver and intestine in 12 months group. Melatonin administration resulted in a significant increase in mean 24 h rSocs1 expression in intestine and heart of 24 months group compared to that of 3 months. The melatonin administration resulted in differential restoration of rSocs1 rhythms and levels in various tissues of 24 months old group. The sensitivity of 24 months old animals to melatonin found in the present study is a step towards endorsing melatonin as an important anti-aging therapeutic drug.

Daily NO rhythms in peripheral clocks in aging male Wistar rats: protective effects of exogenous melatonin.

In mammals suprachiasmatic nucleus (SCN), acts as a light entrainable master clock and by generation of temporal oscillations regulates the peripheral organs acting as autonomous clocks resulting in overt behavioral and physiological rhythms. SCN also controls synthesis and release of melatonin (hormonal message for darkness) from pineal. Nitric Oxide (NO) acts as an important neurotransmitter in generating the phase shifts of circadian rhythms and participates in sleep-wake processes, maintenance of vascular tone as well as signalling and regulating inflammatory processes. Aging is associated with disruption of circadian timing system and decline in endogenous melatonin leading to several physiological disorders. Here we report the effect of aging on NO daily rhythms in various peripheral clocks such as kidney, intestine, liver, heart, lungs and testis. NO levels were measured at zeitgeber time (ZT) 0, 6, 12 and 18 in these tissues using Griess assay in male Wistar rats. Aging resulted in alteration of NO levels as well as phase of NO in both 12 and 24 months groups. Correlation analysis demonstrated loss of stoichiometric interaction between the various peripheral clocks with aging. Age induced alterations in NO daily rhythms were found to be most significant in liver and, interestingly least in lungs. Neurohormone melatonin, an endogenous synchroniser and an antiaging agent decreases with aging. We report further differential restoration with exogenous melatonin administration of age induced alterations in NO daily rhythms and mean levels in kidney, intestine and liver and the stoichiometric interactions between the various peripheral clocks.

Daily rhythms of serotonin metabolism and the expression of clock genes in suprachiasmatic nucleus of rotenone-induced Parkinson's disease male Wistar rat model and effect of melatonin administration.

The circadian system in suprachiasmatic nucleus (SCN) involves regulated serotonin levels and coordinated expression of various clock genes. To understand circadian disfunction in the age-related neurodegenerative disorder Parkinson's disease (PD), the rotenone-induced PD (RIPD) male Wistar rat model was used. The alterations in the rhythmic dynamic equilibrium of interactions between the various components of serotonin metabolism and the molecular clock were measured. There was significant decrease in the mean 24 h levels of tryptophan, 5-hydroxytryptophan (5-HTP), serotonin (5-HT), N-acetyl serotonin (NAS) and melatonin (MEL) by approximately 63, 51, 76 and 96% respectively ( p ≤ 0.05). However significant increase in 5-methoxy indole acetic acid (5-MIAA), 5-methoxy tryptophol (5-MTOH), 5-hydroxy tryptophol (5-HTOH) indicated increased serotonin catabolism with the abolition of daily rhythms of MEL, 5-HTP and 5-MIAA in RIPD. 24 h mean levels of rPer1, rCry1, rBmal1 reduced by about 0.5, 0.74 and 0.39-fold and increased for rPer2 by about 1.7-fold. The daily pulse of rPer2, rCry1, rCry2 and rBmal1 significantly decreased by 0.36, 0.6, 0.14, 0.1 and 0.2-fold. As melatonin, an antioxidant and an endogenous synchronizer of rhythm declined in RIPD male Wistar rat model, the effects of melatonin-administration on the rhythmic expression of various clock genes were studied. Interestingly, melatonin-administration resulted in restoration of the phase of rPer1 daily rhythm in RIPD indicating differential sensitivity of various clock components towards melatonin. The animals which were administered both rotenone and MEL for 48 days interestingly showed neuroprotective effects in dark phase on correlations between expression of various genes.

Melatonin has differential effects on age-induced stoichiometric changes in daily chronomics of serotonin metabolism in SCN of male Wistar rats.

Suprachiasmatic nucleus (SCN) controls various physiological, endocrine and behavioral functions by regulating conversion of serotonin (5-HT) to melatonin (MEL). Aging leads to alterations in the neural and temporal organization of the SCN leading to circadian dysfunction. Age-induced stoichiometric alterations in daily chronomics of various components of 5-HT metabolism were studied by constructing interactomes between parameters. The levels of tryptophan (TRP), 5-hydroxytryptophan (5-HTP), 5-hydroxytryptamine (5-HT), N-acetylserotonin (NAS), N-acetyl 5-methoxytryptamine (MEL), 5-hydroxyindoleacetic acid (5-HIAA), 5-methoxyindole acetic acid (5-MIAA), 5-hydroxytryptophol (5-HTOH), 5-methoxytryptophol (5-MTOH) and N-acetyltryptamine (NAT) were measured at (Zeitgeber time 0, 6, 12 and 18) in male rat SCN of 3, 12 and 24 months age groups. Age-induced decrease was observed in mean levels of NAS, MEL, 5-HIAA, 5-MIAA, 5-MTOH, and NAT and increase was observed in TRP, 5-HTP, 5-HT and 5-HTOH in rat SCN. Daily pulses decreased with aging significantly for TRP, 5-HT, NAS, MEL, 5-HIAA, 5-MIAA and NAT. We report here, the age-induced change in interactions between various 5-HT metabolism components by middle age (12 m) changing further by 24 m. The daily rhythms persisted with aging for NAS, MEL and 5-HTOH. Though, rhythms were abolished for TRP, 5-HTP, 5-HIAA, 5-MIAA, 5-MTOH and NAT differentially at 12 and 24 m. The MEL administration showed restoration in 5-HT ratio with 5-HTP, MEL and 5-HTOH in 24 m and NAS and 5-HIAA in 12 m SCN. Thus, MEL administration effects alterations of age-induced stoichiometry in levels and chronomics of serotonin metabolic network interactomes.

Differential role of melatonin in restoration of age-induced alterations in daily rhythms of expression of various clock genes in suprachiasmatic nucleus of male Wistar rats.

Aging is associated with changes in several basic parameters of circadian rhythms in mammals leading to circadian dysfunction. The hypothalamic Suprachiasmatic nucleus (SCN) regulates neuronal, endocrine and behavioral rhythms through the expression of various clock genes and release of melatonin from pineal gland. In the present study, we investigated the effect of aging on daily rhythms of various clock genes such as rPer1, rPer2, rCry1, rCry2 and rBmal1 in the SCN of male Wistar rats. The m-RNA expression levels of these genes were studied by using quantitative Polymerase Chain Reaction (qPCR) in 3 age groups [3 (adult), 12 and 24 month (m)] at variable time points (Zeitgeber time (ZT)-0, 6, 12 and 18). The m-RNA expression for all genes studied was rhythmic in SCN of adult rats with maximum for rPer1 at ZT-6, rPer2, rCry1 and rCry2 at ZT-12 and rBmal1 at ZT-18. However in 12 and 24 m, the phases of expression of these genes were significantly altered with abolition of daily rhythms of rCry1, rCry2 and rBmal1 in 24 m. Melatonin, messenger of darkness, an endogenous synchronizer of rhythm, an antioxidant and an antiaging drug, declines with aging. We therefore studied the effects of melatonin administered subcutaneously at 1 h before the onset of darkness (ZT-11) for 11 days on age induced desynchronization in expression of these genes. We report here differential restoration of daily rhythm, phase, levels and stoichiometric interaction of m-RNA expression of these genes in various age groups in rat SCN with melatonin treatment.

Effect of restricted feeding on nocturnality and daily leptin rhythms in OVLT in aged male Wistar rats.

Circadian system has direct relevance to the problems of modern lifestyle, shift workers, jet lag etc. To understand non-photic regulation of biological clock, the effects of restricted feeding (RF) on locomotor activity and daily leptin immunoreactivity (ir) rhythms in three age groups [3, 12 and 24 months (m)] of male Wistar rats maintained in light:dark (LD) 12:12 h conditions were studied. Leptin-ir was examined in the suprachiasmatic nucleus (SCN), the medial preoptic area (MPOA) and organum vasculosum of the lamina terminalis (OVLT). Reversal of feeding time due to restricted food availability during daytime resulted in switching of the animals from nocturnality to diurnality with significant increase in day time activity and decrease in night time activity. The RF resulted in % diurnality of approximately 32, 29 and 73 from % nocturnality of 82, 92 and 89 in control rats of 3, 12 and 24 m age, respectively. The increase in such switching from nocturnality to diurnality with restricted feeding was found to be robust in 24 m rats. The OVLT region showed daily leptin-ir rhythms with leptin-ir maximum at ZT-0 in all the three age groups. However leptin-ir levels were minimum at ZT-12 in 3 and 12 m though at ZT-18 in 24 m. In addition the mean leptin-ir levels decreased with increase in food intake and body weight significantly in RF aged rats. Thus we report here differential effects of food entrained regulation in switching nocturnality to diurnality and daily leptin-ir rhythms in OVLT in aged rats.

Identification of specific reference gene for normalization of RT-qPCR data in rhythmic gene expression studies of the effect of developmental hormone antagonist in postembryonic development in Bombyx mori.

Bombyx mori is a lepidopteran holometabolous insect with distinct developmental stages: egg, larvae, pupae, and adult. The lepidopteran insect undergoes major modifications in the central nervous system (CNS) so as to adapt to the lifestyle of these distinct stages with specific habitats and functions from voraciously feeding larval stages to flying reproductive adults via dormant pupal stages. Such transitions are linked to transcriptional, epigenetic, and translational complexities. Therefore, studying rhythmic gene expression in CNS of various developmental stages and the effects of antagonists on developmental hormones requires a very stable reference gene (RG). To facilitate rhythmic gene expression studies using reverse transcription quantitative polymerase chain reaction (RT-qPCR) in B. mori and the effect of developmental hormone juvenile hormone (JH) and 20-hydroxy ecdysone hormone (20 HE), antagonists Precocene 1 and testosterone, respectively, were used. Eight candidate RGs, namely, Translational initiation factor 3 subunit 4 (TI3S4), Translational initiation factor 3 subunit 5 (TI3S5), Ribosomal protein subunit 7 (RPs7), TATA-binding protein association factor (TAF13), Translational initiation factor 4 A (TI4A), Ribosomal protein (RPL32), Elongation factor 1 (EF1), and Arginine kinase (AK), were assessed in the CNS of B. mori. The postembryonic developmental (PED) stages used were the fifth late larval instar, early pupa, mid pupa, late pupa, and adult. The assessments were done at four different time points, Zeitgeber time (ZT) 0, 6, 12, and 18, to find stability towards 24-h rhythmic expression. RefFinder, geNorm, and Ct value analysis were performed. RefFinder and geNORM studies suggested stability order as TI3S4 > TI3S5 > RPs7, but Ct value evaluation showed stability order as TI3S5 > TI3S4 > RPs7. We therefore demonstrated that TI3S4, TI3S5, and RPs7 can be used as RG in various PED stages in CNS of B. mori (Strain: CB-hybrid, PM×CSR2) towards studies with effects of JH and 20 HE antagonists.

Time trends in prostate cancer surgery: data from an Internet-based multicentre database.

OBJECTIVES: To report our experience with an Internet-based multicentre database that enables tumour documentation, as well as the collection of quality-related parameters and follow-up data, in surgically treated patients with prostate cancer. The system was used to assess the quality of prostate cancer surgery and to analyze possible time-dependent trends in the quality of care. PATIENTS AND METHODS: An Internet-based database system enabled a standardized collection of treatment data and clinical findings from the participating urological centres for the years 2005-2009. An analysis was performed aiming to evaluate relevant patient characteristics (age, pathological tumour stage, preoperative International Index of Erectile Function-5 score), intra-operative parameters (operating time, percentage of nerve-sparing operations, complication rate, transfusion rate, number of resected lymph nodes) and postoperative parameters (hospitalization time, re-operation rate, catheter indwelling time). Mean values were calculated and compared for each annual cohort from 2005 to 2008. The overall survival rate was also calculated for a subgroup of the Berlin patients. RESULTS: A total of 914, 1120, 1434 and 1750 patients submitted to radical prostatectomy in 2005, 2006, 2007 and 2008 were documented in the database. The mean age at the time of surgery remained constant (66 years) during the study period. More than half the patients already had erectile dysfunction before surgery (median International Index of Erectile Function-5 score of 19-20). During the observation period, there was a decrease in the percentage of pT2 tumours (1% in 2005; 64% in 2008) and a slight increase in the percentage of patients with lymph node metastases (8% in 2005; 10% in 2008). No time trend was found for the operating time (142-155 min) or the percentage of nerve-sparing operations (72-78% in patients without erectile dysfunction). A decreasing frequency was observed for the parameters: blood transfusions (1.9% in 2005; 0.5% in 2008), postoperative bleeding (2.6%; 1.2%) and re-operations (4.5%; 2.8%). The mean hospitalization time decreased accordingly (10 days in 2005; 8 days in 2008). The examined subcohort had an overall mortality of 1.5% (median follow-up of 3 years). CONCLUSIONS: An Internet-based database system for tumour documentation in patients with prostate cancer enables the collection and assessment of important parameters for the quality of care and outcomes. The participating centres show an improvement in the quality of surgical management, including a reduction of the complication rate.

Melatonin administration differentially affects age-induced alterations in daily rhythms of lipid peroxidation and antioxidant enzymes in male rat liver.

A central clock/pacemaker, suprachiasmatic nuclei of the hypothalamus coordinates and entrains circadian oscillations in the peripheral tissues such as the liver, kidney, heart, lungs etc. called peripheral clocks. These also have endogenous circadian oscillations. The circadian rhythms of antioxidants present in cytosol signify redox state of the cell during day/night cycle. The liver has a major impact on homeostasis through its control on serum protein composition and plays a pivotal role in the metabolism of nutrients, drugs, hormones, and metabolic waste products and undergoes substantial changes in structure and function upon aging. In present study, the temporal patterns of oxidative stress indicators in liver were studied. Daily rhythms of lipid peroxidation end products, reduced glutathione (GSH), oxidized glutathione (GSSG) and antioxidant enzymes such as glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) were studied in liver at variable time points (Zeitgeber Time (ZT) 0, 6, 12 and 18) in three age groups: 3 (adult), 12 and 24 months old male Wistar rats. There was increase in oxidative stress in 12 and 24 months old rats indicated through a significant increase in lipid peroxidation, decrease in GSH/GSSG ratio and antioxidant enzyme activities. In 3 months old rats, lipid peroxidation was maximum at ZT-12 whereas GSH, SOD and CAT activities were minimum at ZT-12. The maximum level in 24 h i.e., acrophases of lipid peroxidation, GPx, SOD and CAT activities in liver cell free extracts altered upon aging. As melatonin, messenger of darkness, an endogenous synchronizer of rhythm, an antioxidant and an antiaging drug, declines with aging we studied the effects of melatonin on activities of these antioxidant enzymes in aging rats. Melatonin administration resulted in differential restoration of acrophases, amplitude, mean as well as daily rhythms of lipid peroxidation and antioxidants in liver of 12 and 24 months old rats.

Effect of melatonin on age induced changes in daily serotonin rhythms in suprachiasmatic nucleus of male Wistar rat.

The decline in physiological functions with aging may affect the ability of the SCN, the biological clock, circadian pacemaker to transmit rhythmic information to other neural target sites, and thereby modify the expression of biological rhythms resulting in circadian disorders. Neurotransmitter serotonin plays important role in the photic and non-photic regulation of circadian rhythms and is a precursor of neurohormone melatonin, an internal zeitgeber. To assess effects of aging on the functional integrity of circadian system, we studied daily serotonin rhythms in the SCN by measuring serotonin levels at variable time points in wide range of age groups such as 15 days, 1, 2, 3 (adult), 4, 6, 9, 12, 18 and 24 months old male wistar rats. Animals were maintained in light-dark conditions (LD; 12:12) two weeks prior to experiment. We report here that in 15 days, 1 and 2 months old rat SCN the mean serotonin level is low and daily serotonin rhythm is just beginning; at 3, 4 and 6 months, serotonin levels and rhythms are robust and at 9, 12, 18 and 24 months mean serotonin levels are low again and rhythm is becoming more disrupted. Previous studies have shown the 5-HT rhythmicity was established by 3 month in rat brain but disintegrated by 6 months of age. As melatonin, an endogenous synchronizer and an antiaging agent, declines with aging, the effects of exogenous melatonin administration on serotonin rhythmicity in SCN in 3, 6, 9 and 24 months old rats were studied to assess effects of aging on responsiveness to melatonin. Our studies indicated an age related loss of sensitivity to melatonin in the restoration of age induced changes in SCN serotonin amplitude and rhythmicity.

Smoking prevention in school students: positive effects of a hospital-based intervention.

BACKGROUND: Adolescents have smoked less in recent years, but 11.7% of 12-to-17-year-olds were still smokers in 2011. The prevalence of smoking has also remained high among 18-to-25-year-olds (36.8%). An intervention program called "Students in the Hospital" was developed in which the health aspects of smoking and its individual and societal consequences were presented in an interactive informational event. In this study, we determine the efficacy of the program. METHODS: From September 2007 to July 2008, we performed an anonymous survey by questionnaire, with a quasi-experimental control-group design, two weeks before (t(1)) and six months after (t(2)) the intervention in a group of 760 participating school students in Berlin. RESULTS: 40.8% of the participants were smokers, among whom 79% stated that they smoked water pipe. Significantly fewer students in the intervention group than in the control group began smoking in the six months after the intervention (p<0.001). The chance of remaining a non-smoker was four times as high in the intervention group (OR, 4.14; CI, 1.66-10.36). Girls benefited from the intervention more than boys (OR 2.56, CI 1.06-6.19). 16.1% of smokers in the intervention group and 17.6% in the control group gave up smoking (p>0.05). CONCLUSION: A clear primary preventive effect of the program was demonstrated, although it apparently did not induce persons who were already smokers to quit.

Immunocytochemical evidence for different patterns in daily rhythms of VIP and AVP peptides in the suprachiasmatic nucleus of diurnal Funambulus palmarum.

The suprachiasmatic nucleus (SCN) is the principal pacemaker that coordinates circadian rhythmicity in mammals. The studies on understanding the circadian system in diurnal rodents are limited. In this study, we have used the 3 striped South Indian Palm Squirrel (Funambulus palmarum). The locomotor activity showed a diurnal pattern of activity in LD 12:12, constant darkness (DD) and light (LL) conditions with circadian periods (τ) of 24.19 ± 0.1, 24.11 ± 0.03 and 24.92 ± 0.35 h respectively. Anatomical study of the brain revealed that this animal had short, thick and stout optic nerves with SCN elliptical in shape with a higher neuronal population as distinct from nocturnal rodents. Since the neuropeptides, vasoactive intestinal polypeptide (VIP) and arginine vasopressin (AVP) play important roles in photic entrainment and relay of information respectively in nocturnal rodents, we studied the distribution and daily rhythms of VIP-ir and AVP-ir in squirrel SCN. The VIP-ir and AVP-ir cells in the SCN showed a ventrolateral and dorsomedial distribution with daily rhythmicity in their levels. The peak time of VIP-ir rhythm was found ahead of AVP-ir. The VIP-ir levels were higher for longer duration than AVP-ir levels. The maximum and minimum VIP-ir levels were at ZT-6 and ZT-0 respectively and AVP-ir levels at ZT-12 and ZT-0 respectively. Thus, VIP and AVP maximum and minimum levels appeared 6 and 12h apart respectively in squirrel, though 12 and 8h apart in rat. These findings in the present report could be a step towards underpinning the mechanisms regulating diurnality.

Daily serotonin rhythms in rat brain during postnatal development and aging.

Aging is characterized by progressive decline in most physiological functions. The age-related sleep disturbances have been attributed to disturbances of circadian function. Neurotransmitter serotonin plays important role in the photic and non-photic regulation of circadian rhythms and is a precursor of melatonin, an internal zeitgeber. To understand the age induced changes in the functional integrity of circadian system, we studied daily serotonin rhythms in brain by measuring serotonin levels at variable time points in wide range of age groups such as 15 days, 1, 2, 3 (adult), 4, 6, 9, 12, 18 and 24-months old male Wistar rats. Animals were maintained under light-dark conditions (LD 12:12), 2 weeks prior to experiment. We report here, mean serotonin levels over 24 h period in brain is highest at 3 months and daily serotonin rhythmicity reliably begins at 3 months and disintegrates at middle age and beyond. The age induced changes in daily serotonin rhythmicity in brain obtained in present study will be a step towards understanding age induced disorders of circadian function.

The effect of curcumin on ethanol induced changes in suprachiasmatic nucleus (SCN) and pineal.

(1) Circadian clocks have been localized to discrete sites within the nervous system of several organisms and in mammals to the suprachiasmatic nucleus (SCN) in the anterior hypothalamus. The SCN controls and regulates the production and discharge of melatonin (hormonal message of darkness) from the pineal gland via a multisynaptic efferent pathway. The nocturnal rise in melatonin production from serotonin results due to an increased activity of serotonin N-acetyl transferase (NAT). (2) The complex interaction between alcohol and biological clock need to be understood as alcoholism results in various clock linked neuronal disorders especially loss of memory and amnesia like state of consciousness, sleep disorders, insomnia, dementia etc. (3) Serotonin, 5-Hydroxy-tryptamine (5-HT) plays an important role in mediating alcohol's effects on the brain. Understanding the impact of alcohol consumption on circadian system is a pre-requisite to help in treatment of alcohol induced neurological disorders. We, therefore, studied the effect of ethanol drinking and ethanol withdrawal on daily rhythms of serotonin and its metabolite, 5-hydroxy-indole acetic acid (5-HIAA) in SCN and Pineal of adult male Wistar rats maintained under light-dark (LD, 12:12) conditions. (4) Curcumin is well known for its protective properties such as antioxidant, anti-carcinogenic, anti-viral and anti-infectious etc. Hence, we studied the effect of curcumin on ethanol induced changes on 5-HT and 5-HIAA levels and rhythms in SCN and Pineal. (5) Ethanol withdrawal could not restore either rhythmicity or phases or levels of 5-HT and 5-HIAA. Curcumin administration resulted in partial restoration of daily 5-HT/5-HIAA ratio, with phase shifts in SCN and in Pineal. Understanding the impact of alcohol consumption on circadian system and the role of herbal medication on alcohol withdrawal will help in treatment of alcohol induced neurological disorders.