Vladimir Kh. Khavinson

Effect of melatonin and pineal peptide preparation epithalamin on life span and free radical oxidation in Drosophila melanogaster.

It was shown previously that epithalamin delays age-related changes in reproductive and immune systems and increases the life span of mice and rats. These effects could be mediated by stimulating influences of epithalamin on synthesis and secretion of melatonin and on free radical processes. A comparative study on the effect of epithalamin and melatonin on both the life span of Drosophila melanogaster (strain HEM) and on the intensity of lipid peroxidation and activity of antioxidative enzymes in their tissues was the main aim of this work. Melatonin and epithalamin was added to the nutrition medium (100 micrograms/ml) during 2-3rd age of larvas. For survival analysis the flies were passed (five coupes per vessel) each 3-7 days. Lipid peroxidation was evaluated as the level of ketodienes (KD) and conjugated hydroperoxides (CHP) in fly tissues at the age of 11 days. Activity of Cu, Zn-superoxide dismuatse (SOD) and catalase was evaluated as well. The mean, median and maximum life span (MLS) were estimated. Mortality rate (MR) was calculated as alpha in the Gompertz equation (R = Ro (exp alpha t) and mortality rate doubling time (MRDT) as in 2/alpha. These parameters in groups of male and female flies exposed to melatonin and in male flies exposed to epithalamin were no different from the parameters for controls. However, exposure to epithalamin was followed in females by a significant increase in mean life span (by 17%, P < 0.02), of median (by 26%), of MLS by 14% and by a 2.12 times decrease of MR (P < 0.01) and MRDT (by 32%) compared with female controls. The level of CHP and KD in the tissues of male control flies was 40 and 49% less than that in females and indirectly correlates with male life span. Exposure to melatonin was followed by a decrease in the level of CHP and KD in females and the deletion of sex differences in them. Exposure to epithalamin significantly decreased the level of CHP and KD in female flies compared to controls (2.3 and 3.4 times, respectively, P < 0.001). Exposure to melatonin failed to influence the activity of catalase in males but increased it in females by 24% (P < 0.02) and failed to influence SOD activity both in males and females. Exposure to epithalamin was followed by a significant increase in activity of catalse, 20% in males and 7% in females and by an increase in SOD activity in males (41%). Thus, it was shown that exposure to epithalamin significantly increases the mean life span and MLS of female D.melanogaster and slowed down their aging rate by 2.12 times. This effect is in good agreement with the inhibiting effect of epithalamin in lipid peroxidation processes in fly tissues.

Pineal peptide preparation epithalamin increases the lifespan of fruit flies, mice and rats

Treatment with pineal peptide preparation epithalamin was followed by the increase of the mean lifespan of female D. melanogaster, SHR mice, C3H/Sn mice and LIO rats by 11-31% (P < 0.05). Ninety percent mortality as well as maximum lifespan were increased in fruit flies, C3H/Sn mice and rats. Mortality rate was decreased by 52% in D. melanogaster, by 52% in rats, by 27% in C3H/Sn mice. It did not change in SHR mice exposed to epithalamin. Treatment with the pineal peptide increased MRDT in flies, C3H/Sn mice and rats. It has been shown that epithalamin increased synthesis and secretion of melatonin in rats and inhibits free radical processes in rats and in D. melanogaster. It is suggested that antioxidative properties of epithalamin lead to increased lifespan of three different animal species.

Effect of synthetic thymic and pineal peptides on biomarkers of ageing, survival and spontaneous tumour incidence in female CBA mice

Fifty female CBA mice were injected s.c. either with 0.1 ml saline, or with synthetic thymic dipeptide Lys-Glu or with synthetic pineal tetrapeptide Ala-Glu-Asp-Gly both in a single dose of 0.1 microg/animal monthly for five consecutive days from the age of 6 months until natural death. Lys-Glu did not significantly influence the body weight and food consumption, free radical processes and estrus function in mice and did increase their physical activity with the subsequent decrease in spontaneous lung adenomas incidence. The pineal peptide treatment was failed to modify the food consumption and physical strength of mice, and was followed by the increase in the body weight, mean survival (by 5.3%, P<0.05) and maximum (by 10 months), by slow down of the ageing of estrus function, by the decrease in body temperature, physical activity, free radical processes and spontaneous tumor incidence (mainly, lung adenomas) in mice. These data suggest the geroprotector potential of the pineal peptide.

Inhibitory effect of the peptide epitalon on the development of spontaneous mammary tumors in HER-2/neu transgenic mice

Female FVB/N HER‐2/neu transgenic mice from the age of 2 months were subcutaneously injected with saline, the peptide Epitalon® (Ala‐Glu‐Asp‐Gly) or with the peptide Vilon® (Lys‐Glu) in a single dose of 1 μg/mouse for 5 consecutive days every month. Epitalon treatment reduced the cumulative number and the maximum size of tumors (p < 0.05). Furthermore, the number of mice bearing 1 mammary tumor was increased, whereas the number of mice bearing 2 or more mammary tumors was reduced in Epitalon‐treated in comparison to saline‐treated animals (p < 0.05). The size but not the number of lung metastases was reduced in Epitalon‐treated compared to saline‐treated mice (p < 0.05). The treatment with Vilon produced significant negative effects when compared to the control group, with an increased incidence of mammary cancer development (p < 0.05), a shorter mean latent period of tumors (p < 0.05) and an increased cumulative number of tumors (p < 0.05). A 3.7‐fold reduction in the expression of HER‐2/neu mRNA was found in mammary tumors from HER‐2/neu transgenic mice treated with Epitalon compared to control animals. The expression of mRNA for HER‐2/neu was also partially reduced in Vilon‐treated mice, but it remained significantly higher in Vilon‐ than in Epitalon‐treated animals (1.9‐fold increase). The data demonstrate the inhibitory effect of Epitalon in the development of spontaneous mammary tumors in HER‐2/neu mice, suggesting that a downregulation of HER‐2/neu gene expression in mammary adenocarcinoma may be responsible, at least in part, for the antitumor effect of the peptide.

Immunomodulatory synthetic dipeptide L-Glu-L-Trp slows down aging and inhibits spontaneous carcinogenesis in rats.

Immunomodulatory molecule L-Glu-L-Trp wasisolated from natural calf thymic peptide complexThymalin by reverse-phase high performance liquidchromatography. On the basis of the synthesizeddipeptide a pharmaceutical was designed containg this compound, which later receives the brand nameThymogen®. The agent activated T-celldifferentiation, T-cell recognition of peptide-MHCcomplexes, induced changes in intracellularcomposition of cyclic nucleotides, and activatedneutrophilic chemotaxis and phagocytosis. The effectof dipeptide on survival, life span and spontaneoustumor development was studied in female rats. Seventy-six, five-month-old outbred female rats were randomlysubdivided into two groups and were subcutaneouslyinjected with 0.2 ml of normal saline (controls, 32 rats) or with 5 µg/rat of the dipeptideL-Glu-L-Trp, dissolved in 0.2 ml of saline (44 rats),5 times per week for 12 months. Animals were monitoredup to their natural death and all the tumorsdiscovered were studied microscopically. Mean lifespan of rats in both groups was similar but that of10% maximum survived control rats constituted949 ± 16.1 days, whereas in the dipeptide-treatedrats this value was 1048 ± 21.1 days (P < 0.001).Six out of 44 rats treated with the drug survived overthe maximum life span of control rats (965 days). Theaging rate indicated as α in the Gompertz equation,was 0.0071 days−1 in controls and 0.0041 days−1 in rats exposed to L-Glu-L-Trp. Totaltumor incidence was 1.5 times lower (P < 0.01),malignant tumor incidence 1.7 times lower (P < 0.01),and hematopoietic malignancies (leukemias andlymphomas) 3.4 times lower (P < 0.02) in rats exposedto the dipeptide in comparison with controls. Thus,treatment with L-Glu-L-Trp delayed aging rate anddecreased spontaneous tumor incidence in rats.

Effect of Epitalon on biomarkers of aging, life span and spontaneous tumor incidence in female Swiss-derived SHR mice

From the age of 3 months until their natural deaths, female outbred Swiss-derived SHR mice were subcutaneously injected on 5 consecutive days every month with 0.1 ml of normal saline (control) or with 1.0 µg/mouse (∼30–40 µg/kg) of tetrapeptide Epitalon®(Ala-Glu-Asp-Gly) dissolved in 0.1 ml saline. There were 54 mice in each group. The results of this study show that treatment with Epitalon did not influence food consumption, body weight or mean life span of mice. However, it slowed down the age-related switching-off of estrous function and decreased the frequency of chromosome aberrations in bone marrow cells (by 17.1%, P < 0.05). It also increased by 13.3% the life span of the last 10% of the survivors (P < 0.01) and by 12.3% the maximum life span in comparison with the control group. We also found that treatment with Epitalon did not influence total spontaneous tumor incidence, but inhibited the development of leukemia (6.0-fold), as compared with the control group. The data obtained suggest a geroprotector activity of Epitalon and the safety of its long-term administration in mice.

Experimental Studies of the Pineal Gland Preparation Epithalamin

Twenty-five years of study have shown a wide spectrum of high biological activity of the pineal peptide preparation epithalamin. Long-term exposure to epithalamin was followed by an increase in the mean and maximum life spans and slower rates of aging of rats, mice, and D. melanogaster. Epithalamin increases pineal synthesis of serotonin, N-acetylserotonin, and melatonin and night pineal secretion of melatonin in adult and old rats. The pineal preparation decreases the luteinizing hormone and prolactin levels in adult male rats as well as the threshold of the hypothalamopituitary complex to feedback inhibition by estrogens in old female rats; it slows dawn age-related cessation of estrous function in rats and induces the recurrence of estrous cydes and fertility in old, persistently estrous rats. Epithalamin increases the levels of triiodothyronine and decreases thyroxine in serum of adult rats. It further decreases the levels of corticosterone in the serum of mice and increases the susceptibility of the hypothalamo-pituitary complex to the homeostatic inhibition of adrenocorticotropic function by glucocorticoids in old rats. Serum insulin and triglyceride levels in rabbits are decreased by epithalamin and the tolerance to glucose and diuresis are increased. With respect to immune function, it was found that T and B cell-mediated immunity in adult and old mice as well as the titer of thymic serum factor and the titer of thymosin-like compounds in old mice are stimulated by the pineal peptide preparation in the same way as the colony-forming activity of splenocytes in pinealectomized rats. Epithalamin inhibits spontaneous and induced carcinogenesis and is a potent antioxidant, decreasing lipid peroxidation and stimulating the activity of CuZn superoxide dismutase. The obtained results demonstrate a high efficiency of epithalamin therapy for prophylaxis of age-related pathology, including cancer, showing a new physiological way to slow down pathological processes and to extend human life spans.

Inhibitory effect of peptide Epitalon on colon carcinogenesis induced by 1,2-dimethylhydrazine in rats

The effect of synthetic pineal peptide Epitalon (Ala-Glu-Asp-Gly) on colon carcinogenesis was firstly studied in rats. Eighty 2-month-old outbred male LIO rats were subdivided into four groups and were weekly exposed to five subcutaneous injections of 1,2-dimethylhydrazine (DMH) at a single dose of 21 mg/kg body weight. Additionally, 5 days a week, some of the rats were given subcutaneous injections of saline at a dose of 0.1 ml during the whole experiment (group 1, control) or Epitalon at a single dose of 1 microg during the whole experiment (group 2), Epitalon after termination of carcinogen injections (group 3) or during the period of DMH exposure (group 4). Colon carcinomas developed in 90-100% of DMH-treated rats. The number of total colon tumors per rat was 4.1; 2.7; 3.7; 2.9 in groups 1, 2, 3, 4, respectively (the difference in groups 2 and 4 compared with group 1 is significant). In rats from group 2, colon tumors were smaller than in control animals. In group 2, the incidence, as well the multiplicity of tumors in ascending and descending colon, were significantly decreased in comparison with group 1. In group 4, the mean number of tumors per rat was significantly decreased, too. A trend to decrease the number of tumors in the rectum in rats from groups 2, 3 and 4, treated with Epitalon was found. Epitalon inhibited also the development of tumors in jejunum and ileum. Thus, our results demonstrated an inhibitory effect of Epitalon on chemically induced bowel carcinogenesis in rats.

Biogerontology in Russia: from past to future

The paper presents major steps of gerontology development in Russia. The issues of training in gerontology and geriatrics, institutional infrastructure within the Gerontological Society of the Russian Academy of Sciences and its activities have been considered therein. Some results of Russian researchers obtained during 2005–2010 have been summarized as well. Special attention is given to the prospects of gerontology in Russia.

Biogerontological research in Europe: special issue.

As the current President and the Secretary, respectively, of the Biological Section of the International Association of Gerontology and Geriatrics (IAGG; European Region), we are delighted to present this special issue of Biogerontology on the occasion of the 7th European Congress of IAGG at Bologna (April 14–17, 2011), providing an overview of the state of biogerontological research in various countries in Europe. Research on the biological basis of ageing includes: (1) describing the phenotype of ageing at the level of organs, systems, tissues, cells, intraand inter-cellular pathways, and molecules; (2) unravelling the biochemical and molecular mechanisms of age-related changes; (3) identifying genes that affect the quality and duration of lifespan; (4) identifying the rate-limiting steps which lead to the emergence of age-related diseases; and (5) screening, testing and developing evidence-based effective interventions to modulate ageing and to extend the health-span. Different countries have different priority areas for research, which keep on changing and evolving in accordance with the changing social, political and economic trends. There are 34 national gerontological research societies in Europe, and they are expected to maintain databases with respect to all aspects of ageing research, including biological, sociological, psychological, and clinical. From about mid-2009, we approached them with an invitation to present a brief overview of the state of biogerontological research in their respective country. We are delighted to present the first series of such reports from Austria, Czech Republic, Finland, Georgia, Israel, Italy, Russia, Serbia, Spain, Turkey and Ukraine in this special issue of Biogerontology, and we hope that similar reports from the other countries will be published in the future issues.