Current Aging Science (v.5, #3)

Light-at-Night-Induced Circadian Disruption, Cancer and Aging by Vladimir N. Anisimov, Irina A. Vinogradova, Andrei V. Panchenko, Irina G. Popovich, Mark A. Zabezhinski (170-177).
Light-at-night has become an increasing and essential part of the modern lifestyle and leads to a number of health problems, including excessive body mass index, cardiovascular diseases, diabetes, and cancer. The International Agency for Research on Cancer (IARC) Working Group concluded that “shift-work that involves circadian disruption is probably carcinogenic to humans” (Group 2A) [1]. According to the circadian disruption hypothesis, light-at-night might disrupt the endogenous circadian rhythm and specifically suppress nocturnal production of the pineal hormone melatonin and its secretion into the blood. We evaluated the effect of various light/dark regimens on the survival, life span, and spontaneous and chemical carcinogenesis in rodents. Exposure to constant illumination was followed by accelerated aging and enhanced spontaneous tumorigenesis in female CBA and transgenic HER-2/neu mice. In male and female rats maintained at various light/dark regimens (standard 12:12 light/dark [LD], the natural light [NL] of northwestern Russia, constant light [LL], and constant darkness [DD]) from the age of 25 days until natural death, it was found that exposure to NL and LL regimens accelerated age-related switch-off of the estrous function (in females), induced development of metabolic syndrome and spontaneous tumorigenesis, and shortened life span both in male and females rats compared to the standard LD regimen. Melatonin given in nocturnal drinking water prevented the adverse effect of the constant illumination (LL) and natural light (NL) regimens on the homeostasis, life span, and tumor development both in mice and rats. The exposure to the LL regimen accelerated colon carcinogenesis induced by 1,2-dimethylhydrazine (DMH) in rats, whereas the treatment with melatonin alleviated the effects of LL. The maintenance of rats at the DD regimen inhibited DMH-induced carcinogenesis. The LL regimen accelerated, whereas the DD regimen inhibited both mammary carcinogenesis induced by N-nitrosomethylurea and transplacental carcinogenesis induced by N-nitrosoethylurea in rats. Treatment with melatonin prevented premature aging and tumorigenesis in rodents. The data found in the literature and our observations suggest that the use of melatonin would be effective for cancer prevention in humans at risk as a result of light pollution.

Melatonin and Pineal Gland Peptides Are Able to Correct the Impairment of Reproductive Cycles in Rats by Alexander Arutjunyan, Ljudmila Kozina, Yulia Milyutina, Andrew Korenevsky, Michael Stepanov, Vladimir Arutyunov (178-185).
Catecholamines play an important role in the hypothalamic regulation of the synthesis and secretion of gonadotropin- releasing hormone, or gonadoliberin. We have shown that melatonin and the pineal gland peptides (epithalamine and epitalon) exert a correcting influence on the diurnal dynamics of norepinephrine (NE) in the medial preoptic area (MPA) and of dopamine (DA) in the median eminence with arcuate nuclei (ME-Arc) disturbed by single administration of the neurotoxic xenobiotic 1,2-dimethylhydrazine (DMH) in female rats. It has been found that experiments with DMH administration can be used as an animal model of female reproductive system premature aging. The investigation of epithalamine (a polypeptide preparation from the bovine pineal gland) effect on circadian rhythms disturbed by the neurotoxic compound DMH has shown a recovery of the diurnal dynamics of NE in MPA. In addition, NE was found to decrease from 9:30 till 11 o’clock, Circadian Time (CT), which was typical of control animals. Epitalon (Ala-Glu-Asp-Gly) proved to be more effective in ME-Arc. This peptide prevents the xenobiotic caused disturbance of DA diurnal rhythm, keeping this metabolite low at 5 o’clock (CT) with it having increased by 11 o’clock (CT). The data obtained suggest that the pineal gland is important for the circadian signal normalization needed for gonadoliberin surge on the day of proestrus. Melatonin and peptides of the pineal gland can be considered as effective protectors of female reproductive system from xenobiotics and premature aging.

Longitudinal Melatonin Production in Female Laboratory Rats During 1997-2006: Possible Modulatory Effects of Changing Solar Activity by Hella Bartsch, Dieter Mecke, Hansgeorg Probst, Heinz Kupper, Eckhard Seebald, Ilse Herget, Elmar Peschke, Lothar Salewski, Thilo Stehle, Christian Bartsch (186-194).
Earlier we reported that the urinary excretion of 6-sulfatoxymelatonin (aMT6s) displayed seasonal rhythms in laboratory rats and hypothesized that the horizontal intensity H of the geomagnetic field may act as seasonal zeitgeber. To test this, long-term experiments were performed with female Sprague-Dawley rats. In experiment I (n=12: 1997-1999) nocturnal aMT6s displayed a winter-summer increase by 30% and a rhythm with a phase-length of 24 months peaking in July 1998. In experiment II (n=12; 1999-2000) the winter-summer increase amounted to 40%. The estimated rhythm had a phase-length of 18 months with a peak in September 2000. Compared to experiment I both the rhythm-adjusted mean (MESOR, + 28%) and amplitude (+68%) were elevated. In experiment III (n=30; 2003-2004) the winter-summer increment was just 20%. A circannual rhythm with a peak in April/May was found. The MESOR was 13% higher than in experiment I but the amplitude was depleted ( -14%). In experiment IV (n=15; 2005-2006) a slight winter-summer increase (+15%) was found and a low-amplitude rhythm of 24 months phase-length peaking in June 2006. The MESOR was similar to experiment I but the amplitude was depressed (-36%). These results demonstrate that female rats within two years of age show elevated aMT6s during summer/spring which supports our initial hypothesis. The apparent inter-experimental amplitude variation indicates the involvement of additional variables. Based on our initial hypothesis, we postulate an involvement of the solar cycle affecting H leading to year to year variations and present supportive analyses.

Aging and aging related illnesses such as cancer have been associated with inflammatory changes. Cancerrelated behavioral comorbidities such as fatigue, sleep disturbances, depression have also been associated with inflammation, hypothalamic-pituitary-adrenal (HPA) axis dysregulation and other neuroendocrine changes. From a clinical perspective, cancer-related fatigue demonstrates striking similarities with the cytokine-induced sickness phenomenon in animal models. Thyrotopin-releasing hormone (TRH) plays a homeostatic role in its interaction with several biological systems, including a critical role in its interactions with the immune system. Considerable evidence supports a pivotal role for TRH in the inflammatory processes with specific relevance to the “cytokine-induced sickness behavior” paradigm. Additionally, TRH exerts arousing and analeptic effects in instances of behavioral depression. In a small proof-of-concept study conducted by our group, we investigated TRH administration as a treatment fatigue in cancer survivors in comparison with saline administration using a double-blind, crossover design. We also evaluated impact of TRH/saline administration on the inflammatory markers in these patients. TRH administration was associated with significant improvement (p < 0.05) in fatigue levels as measured by the Visual Analog Scale-Energy (VAS-E), was associated with significant (p < 0.05) improvement in sleep disturbances and improved quality of life. Notably, TRH administration was associated with decrease in C-reactive protein (CRP) levels, a marker of inflammation. This decrease in CRP level with TRH administration was associated with improvement in energy levels as measured by the VAS-E. The present review supports potential utility of TRH-based therapeutics in medical and psychiatric disorders with underlying inflammatory processes.

Telomere Shortening Is a Sole Mechanism of Aging in Mammals by Victor M. Mikhelson, Irina A. Gamaley (203-208).
We adduce proof that telomere shortening is the sole mechanism of aging. All apparent contradictions, particularly the absence of an inverse correlation between residual telomere length and donor age, are explained within the bounds of telomere theory. We explain in what way telomere shortening might be the cause of aging and lifespan restriction. We also show the inability of the oxidative theory to explain a number of indisputable (and easily explained by telomere theory) facts, such as malignant growth of tumor cells and why children begin aging not from the level reached by the cells of their parents at the moment of conception but from nothing. We postulate that if oxidative damage was entirely absent, telomeres would, nevertheless, shorten with each mitotic cycle because this is the mechanism of DNA replication. Aging would occur all the same, and it is the very thing we can observe under the effect of any antioxidants. If telomeres do not shorten, as is the case in transformed cells in which telomerase is working, aging will do stop and transformed cells will show no senescence. We also observe this in spite of the damaging effect of reactive oxygen species, which is even more intensive in transformed cells than in normal cells.

In the central nervous system (CNS) microglia are crucial for the defense of the brain against invading microorganisms, formation of tumors, and damage following trauma [1]. However, uncontrolled activation of these cells may have deleterious outcomes [2] through activation of Fcγ and the complement 3 receptors and the induction of an adaptive immune reaction [3]. Proteins contributing to this reaction are the intercellular adhesion molecule-1 (ICAM-1) [3] and CD3 molecules, among others. Both can be expressed on the glia cells before cytokine release and may facilitate an autoimmune inflammatory reaction in the brain. Round microglial cells among the pyramidal cells of the hippocampus with increased expression of CD32+ (FcγIIa) and near the site of injection of aluminum were detected immunohistochemically and indicate microglial activation at the site of aluminum injury. ICAM-1+ immunoreactivity significantly increased in the hippocampus and in the choroids plexus, indicating increased inflammation in the brain as well as increased CD3&#x3BE;+ expression in the hippocampus and non-MHC-restricted T cytotoxicity after aluminum injection. The pattern of expression of CD32+ (FcγIIa receptor) near the site of aluminum injection indicates that microglia may play a phagocytic role at the site of aluminum-induced excitotoxicity in the brain. Significant expression of ICAM-1+ and CD3&#x3BE;+ immunoreactive cells with the clusters of ICAM-1+ in the choroid plexus suggests a consequently neurotoxic autoimmune reaction induced by microglial hyperactivation in the injured brain.

We are entering a post-antibiotic era in medicine, as resistance to commonly used antibiotics rises and the discovery of successful new classes of antibiotics slows. New therapeutic targets are being identified and investigated, including approaches that may overcome resistance, interrupt bacterial communication, and enhance human immune function. Immune function is negatively affected by type B malnutrition (multiple micronutrient depletion), “pathological hygiene” (removal of innate immune primers such as yeasts from the food chain), the removal of innate primers such as yeasts from the food chain, HIV infection, diabetes, chronic stress and depressive illness, and the use of immunosuppressant medications. These and other forms of innate immune dysfunction can be overcome / compensated for by novel therapeutic tools which enhance or mimic different aspects of innate immune function, and which re-configure and reinforce Claude Bernard’s milieu interieur.

Receptor-mediated Oxidative Stress in Murine Cerebellar Neurons is Accompanied by Phosphorylation of MAP (ERK 1/2) Kinase by Yulia Rybakova, Evgeny Akkuratov, Konstantin Kulebyakin, Olga Brodskaya, Antonina Dizhevskaya, Alexander Boldyrev (225-230).
A primary culture of murine cerebellar neurons was used to induce oxidative stress resulting in the accumulation of reactive oxygen species (ROS) and activation of ERK 1/2 kinase. Short-term incubation (15 min) of cerebellar neurons with homocysteine (HC) or N-methyl-D-aspartate (NMDA) induced partial ERK 1/2 phosphorylation thus providing the activation of the enzyme. Inhibitors of NMDA receptors, MK-801 or D-AP5, both prevented the activation of cells by HC or NMDA. Another receptor-dependent means of oxidative stress stimulation is exposure of cells to the cardiac glycoside ouabain, a specific inhibitor of Na/K-ATPase. Ouabain induces ROS accumulation and substantial ERK1/2 activation in neuronal cells at concentrations as low as 1 nM - 1 M, which corresponds to participation of Na/K-ATPase in intracellular signalling. Neuropeptide carnosine added to the cells 2 hours before oxidative stress prevented both ROS accumulation and ERK1/2 activation. As ERK1/2 kinase plays a key role in gene expression responsible for either cell adaptation or cell death, the model used gives a useful tool to characterize the effect of natural and synthetic anti-cancer drugs on cellular life. The data presented show that carnosine is a natural modulator of oxidative stress in neuronal cells, providing regulation of ERK1/2 activity via buffering intracellular ROS levels.

Aging and advanced cancer are characterized by similar neuroendocrine and immune deficiencies; the most important of them consist of diminished nocturnal production of the pineal hormone melatonin (MLT) and decreased production of IL-2. At present, however, it is known that the pineal gland may produce indole hormones other than MLT. The most investigated of them is represented by 5-methoxy-tryptamine (5-MTT), which may exert antitumor, anticachectic, and immunomodulating effects under experimental conditions, in addition to those effects produced by MLT itself. In an attempt to obtain some preliminary data in human subjects about the potential therapeutic properties of 5-MTT, three different studies of 5-MTT have been carried out in advanced solid tumor patients. The first study of MLT plus 5-MTT included 14 thrombocytopenic cancer patients who did not respond to MLT alone. In the second study we have compared the clinical efficacy of MLT plus 5-MTT in a group of 25 untreatable metastatic cancer patients to the results obtained in a control group of 25 cancer patients receiving MLT alone. Finally, the third study of MLT plus 5-MTT included 14 untreatable metastatic cancer patients who did not respond to MLT alone. In all of these studies, MLT and 5-MTT were given orally at the level of 20 mg/day in the evening and at 5 mg/day during the period of maximum light. A normalization of platelet number was achieved by MLT plus 5-MTT in 5 of 14 (36%) thrombocytopenic cancer patients who did not respond to MLT alone. The percentage of disease control obtained by MLT plus 5-MTT in untreatable metastatic cancer patients was significantly higher than that achieved by MLT alone (15/25 [60%] vs. 8/25 [32%], P < 0.05). Finally, the association of 5-MTT with MLT induced disease stabilization in 4 of 14 (29%) untreatable metastatic cancer patients who did not respond to MLT alone.

Natural Dipeptides as Mini-Chaperones: Molecular Mechanism of Inhibition of Lens βL-Crystallin Aggregation by Antonina K. Dizhevskaya, Konstantin O. Muranov, Alexander A. Boldyrev, Mikhail A. Ostrovsky (236-241).
The effect of histidine-containing dipeptides—carnosine and N-acetylcarnosine—on preventing and treating of cataracts of various etiologic origins has been demonstrated in many studies in vivo, while the precise molecular mechanism of their action is actually obscure. Cataract has been recently attributed to conformational diseases due to the association of lens structure protein aggregation with cataract pathogenesis. In our study, effect of histidine-containing dipeptides—carnosine, N-acetylcarnosine, and anserine—on the UV induced βL-crystallin aggregation was studied in vitro. It was first demonstrated that N-acetylcarnosine and anserine (10-40 mM) considerably suppressed UV induced aggregation of βL-crystallin, while carnosine exerted no effect. Positive correlation between anti-aggregating activity of the compounds used and their hydrophobicity was obtained. It was revealed that N-acetylcarnosine and anserine inhibited the initial stages of the protein photochemical damage. A decrease in the size of protein aggregates was detected in the presence of N-acetylcarnosine and anserine. UV irradiation of βL-crystallin resulted in a significant increase in the number of protein carbonyl groups, and the dipeptides studied did not affect this process. We suppose that N-acetylcarnosine and anserine inhibit βL-crystallin aggregation via formation of a protein-dipeptide complex that prevents macromolecular conformational changes and ensuing protein aggregation.

Effect of Chronic Melatonin Administration on Several Physiological Parameters from Old Wistar Rats and Samp8 Mice by Jesus A.F. Tresguerres, Roman Kireev, Katherine Forman, Sara Cuesta, Ana F Tresguerres, Elena Vara (242-253).
The effect of melatonin administration on age-induced alterations in hepatocytes, central nervous system, immune system, and skin are reviewed. Twenty-two-month-old Wistar rats and SAMP8 (senescence prone) mice of 10 months of age were used as experimental models. Wistar rats were analyzed untreated or after the chronic administration of melatonin at a dose of 1 mg/kg/day in the drinking water for 10 weeks. At the end of the treatment period, the various parameters were investigated. Results were compared with those of 2-month-old controls. In hepatocytes, aging induced a significant increase in oxidative stress, inflammation, and apoptosis when compared to young animals. Melatonin administration significantly ameliorated all these age-related changes. The impairment of the cardiovascular system with aging appears to contribute to the increased morbidity and mortality of the aged subjects. The process was investigated in SAMP8 mice of 10 months of age. Melatonin was provided for 30 days at two different dosages (1 mg/kg/day and 10 mg/kg/day), also in the drinking water. After treatment, the expression of inflammatory mediators (tumor necrosis factor-α, interleukin 1 and 10, NF&#x3BA;Bp50 and NF&#x3BA;Bp52), apoptosis markers (BAD, BAX, and Bcl2), and parameters related to oxidative stress (heme oxygenases 1 and 2, endothelial and inducible nitric oxide synthases) were determined in the heart. Inflammation as well as oxidative stress and apoptosis markers were increased in old SAMP8 males, as compared to young controls. After treatment with melatonin, these age-altered parameters were partially reversed. The results suggest that oxidative stress and inflammation increase with aging and that chronic treatment with melatonin is able to reduce these parameters. In the skin, a reduction of epidermal thickness together with a marked increase of the hypodermis with great fat accumulation was observed in old rats, together with an increase in caspase 3, 8 of nucleosomes and LPO and a reduction in Bcl2 levels in the cultured keratinocytes. Melatonin treatment was able to reduce the fat content of the hypodermis and to increase Bcl2 and reduce nucleosomes, caspases, and LPO in keratinocytes. Conclusion: Melatonin administration exerts a beneficial effect against age-induced changes in several physiological parameters in Wistar rats and SAMP 8 mice.

Heat Shock Protein Gp96 as Potential Regulator of Morphostasis after Partial Hepatectomy in Mice by Biserka Radosevic-Stasic, Hrvoje Jakovac, Damir Grebic, Zlatko Trobonjaca, Ines Mrakovcic- Sutic, Mira Cuk (254-262).
Gp96 (also known as glucose-regulated protein 94, endoplasmin) is the endoplasmic reticulum (ER)-resident protein, which belongs to the heat shock protein HSP90 family. It is upregulated in response to glucose starvation and other stressful stimuli that disrupt protein synthesis in the ER. There, it is acting as a molecular chaperon involved in the correction of unfolded proteins, in the activation of proteasome-dependent ER-associated degradation of the misfolded proteins, and in activation of protein translations that modulate the polypeptide traffic into the ER. In addition, it has been implicated in antigen presentation and MHC class I and II upregulation, in the activation and maturation of dendritic cells and proinflammatory cytokine secretion, as well as in chaperoning of integrins and Toll-like receptors, acting as a "danger signal" to the innate and adaptive immunity. Moreover, owing to its specific function in Ca2+ homeostasis and in the insulin– IGF/signaling pathways, it has been proposed that gp96 might participate in mechanisms that are critical for cell growth, differentiation, and responses to ER stress. Emphasizing that gp96, as a natural adjuvant for chaperoning antigenic self peptides into the immune surveillance pathways, may also be involved in the maintenance of morphostasis and self tolerance, in this survey we show that high levels of upregulation of gp96 in regenerating liver and thymus are followed by signs of transient autoimmunity, augmented apoptosis in thymus, and the presence of autoreactive NKT and regulatory T cells that might be involved in the control of rapid liver growth induced by partial hepatectomy.

Psychoimmunological Analysis of Cancer Patients: Correlation with the Prognosis by Giuseppina Messina, Paolo Lissoni, Franco Rovelli (263-272).
Thanks to the discoveries of psychoneuroendocrinoimmunology, we now know that every psychological state is mediated by a specific neurochemical condition and every neurochemical change in turn influences psychological status. We can now identify three different levels of neurochemical mediation of the psychological states: neurotransmission, neuromodulation, and the psychoneuromodulation. Neurotransmission is composed of five main neural pathways, noradrenaline, acetylcholine, dopamine, serotonin, and histamine; neuromodulation; and the psychoneuromodulation. We have performed several clinical studies in an attempt to correlate the psychological status of cancer patients with the immune alterations characteristic of the clinical history of neoplastic disease. We have studied the immunologic status by evaluating cytokine blood levels and the lymphocyte subpopulation; the psychological status was assessed by the Rorschach's test; and spiritual status was evaluated by a previously published test to explore spiritual faith. These preliminary psychological studies seem to suggest that a pre-treatment analysis of psychological and spiritual status may predict the efficacy of both chemotherapy and immunotherapy in advanced cancer patients.

Self-Repair in Degenerative Joint Disease by Valerio Di Nicola, Renato Di Nicola (273-287).
This study presents a method for treating and structurally improving articulations affected by degenerative joint disease (DJD). The focus of this analysis is on two groups of patients: the first comprised patients over eighty years old, and the second comprised patients aged 45 to 55 years. The first group was a high surgical risk and both had been nonresponders to current conservative therapies. Scholars like Davis, Filatov, and Cerletti have been studying and using the regenerative properties of placenta, amnios and other nonvital tissues since the early 1900s. These pioneering studies have opened a new track for tissue renewal. More recently, the new biological knowledge about extracellular nucleic acids, growth factors (GF) (as by-products of trauma response), and heat shock proteins (Hsp) has helped research even further. Building on those experiences, we have developed a regenerative gel obtained with distressed, processed blood, polydeoxyribonucleotides (Pdrn), and a thickening substance. The objective was to stimulate the local innate stem cells with our gel in order induce tissue repair. From 2003 until 2009, we treated 948 patients. As mentioned, the first group comprided of 86 ultra-octogenarian patients with severe osteoarthritis (OA) of the hip and/or knee, and the second group comprised of 90 younger patients (around 50 years old) affected by the same disease. Treated patients have been clinically and radiologically evaluated with a follow-up of 6 to 48 months. Results show a statistically significant improvement in terms of pain and joint mobility, sometimes coupled with clear improvement in radiological imaging. Follow-up shows encouraging data in terms of clinical stability over time. During the study, we encountered virtually no side effects, adverse reactions, or toxicity. Currently the pharmacological treatment of DJD is palliative, though toxicity and side effects of the drugs remain problematic. Patients who can be operated on conclude their trial with a prosthesis followed by a long rehabilitation period. This study presents a new methodological approach to the treatment of DJD based on tissue regeneration and restoration resulting in a positive clinical resolution.