[Melatonin effects in young and aged mice with toxic cuprizone-induced model of demyelination.]

The followings were estimated in the 3-5 and 15-17 months 129/Sv cuprizone- and melatonin-treated mice: the number of activated T-lymphocytes, macrophages, neural stem cells (determined by CD3+, Mac1+ and nestin+ markers), the structurally unchanged neurons, the malondialdehyde (MDA) content and the antioxidant enzyme activities in the brain; the blood thymus hormone thymulin level; and the behavioural indices. The mice were fed with cuprizone for 3 weeks. From the 8th day of the cuprizone treatment the mice were injected with melatonin (1 mg/kg, at 18:00 daily). As a result, the number of the CD3+-, Mac1+-, and nestin+-cells and the MDA content increased while the glutathione peroxidise (GP) activity decreased in the brain of young and aged mice under the influence of cuprizone. In mice of both age groups the proportion of unchanged neurons in the central nervous system, motor and emotional activity and muscle tone were decreased. Regardless of the age of the mice the injections of melatonin decrease the number of СD3+, Мас1+-cells, content of MDA, increase activity of GP and thymulin level. Decrease of the number of nestin+-cells coincides with the growth of the number of unchanged neurons. The effect of both neurotoxin and melatonin on immune factors, the structure and functional state of neurons was more pronounced in young mice. Thus, the positive effect of melatonin in young and aged mice with the cuprizone-induced demyelination was realized mainly through the pathogenetic factors of pathology.

[THYMIC HORMONES, ANTIOXIDANT ENZYMES AND NEUROGENESIS OF BULBUS OLFACTORIUS IN RATS WITH PARKINSONISM: THE EFFECT OF MELATONIN].

The adult rats received both neurotoxin 6-hidroxidophamine and neurotoxin and melatonin. It was investigated a link between the disturbances of the brain antioxidant enzymes activity and thymic endocrine function, as possible pathogenic factors of parkinsonism, with changes in the number of neural stem cells (NSC) in the bulbus olfactorius. Rats with motor asymmetry in the apomorphine test and significant damage of the dopaminergic neurons in the-substantia nigra have decreased levels of superoxide dismutase, catalase and glutathione peroxidase activities in striatum (1.3-1.4 times) and blood thymulin content (8 times) compared to control group. On the contrary, examined indices were not changed in rats without motor asymmetry and correspondingly partly damaged neurons. The number of nestin(+)-cells in the bulbus olfactorius of rats without motor asymmetry increased from 91.2% to 99.3% and remained unchanged after melatonin administration course (10 mg/kg during 18 days). Melatonin administration resulted in the decrease in the number of nestin(+)-cells along with significant elevation of the decreased antioxidant enzymes activity and blood thymulin content in rats with circulatory movements. Possibilities of the enhancement of NSC differentiation in bulbus olfactorius into neuronal direction in such animals has been discussed. The conclusion about the potential use of melatonin as a neuroprotector in parkinsonism therapy has been made.

[The sexual peculiarities of aging changes in circannual rhythms of pineal gland, hypophysis, adrenal cortex and thymus functions in healthy subjects].

The interrelations of circannual rhythms of the functional state of pineal gland, hypophysis, adrenal cortex, thymus in healthy women and men from 20 to 79 years were studied. Fluctuations of melatonin, ACTH, cortisol and thymic serum factor, which were exchanged in aging (the season peaks of hormones and its acrophase) were found in blood of healthy 20-29 years old people. The changes of rhythmicity of indices were in male earlier (pineal gland and hypophysis over 30 years, thymus and adrenal cortex over 40 years) and more impressive than in women. The aging changes of pineal gland function's rhythm in healthy subjects have important role for changes of interrelations of circannual rhythms hypophysis, adrenal cortex and thymus.

[The aging particularities of bone marrow composition, pineal gland and thymus functions in mice of different lines].

We investigated the amount of stromal precursor cells for colonies of fibroblasts (CFC-F) and progenitor cells for granulocyte-macrophage colonies (CFC-GM cells), blood content of thymulin and melatonin in bone marrow of young and old mice CBA/Ca and FVB/N lines. The CBA/Ca mice demonstrated only weak increasing amount of CFC-F and CFC-GM in bone marrow, but these indices in FVB/N mice are increased more significantly. Linear difference of age-related changes in the biological features of the cells of bone marrow are significantly associated with the characteristics and relationships of the function of epiphysis and the thymus in mice of different lines during aging.

[Thymus and adaptive changes of bone marrow cell composition in animals of different age].

The influence of thymus removal or administration of thymalin, thymic peptide factor, on number of cells-precursors for colonies of fibroblasts (CFC-F) and for granulocyte-macrophageal colonies (CFC-GM), CD(4+)-cells in bone marrow in CBA mice of different age in some seasons was investigated. It was shown that in adult mice the orientation of influence thymic activity modulation on amount of these types cells is dependent from seasons. It was shown the season possibility of direct effect of thymic stroma supernatant, in which was thymic serum factor (FTS), or the synthetic FTS on the amount of CFC-F and CFC-GM in bone marrow of adult mice; in old mice thymic supernatant increased in vitro the amount of CD(4+)-cells. FTS level in old mice increased after administration of thymalin, the rhythm of CD(4+)-cells number in bone marrow and of corticosterone serum content restored.

[Influence of the pineal gland's factors on the cell composition of the bone marrow in animals of different age in conditions of changed thymus functional state].

The role of the thymus in the influence of pineal gland on the number of stromal precursor cells to colonies of fibroblasts (CFU-F), progenitor cells to granulocyte-macrophage colonies (CFU-GM), CD3+, CD4+ and CD8+ cells in the bone marrow of adult and old mice CBA/Ca was investigated. False-operated and thymusectomized mice were injected Melatonin in the evening (spring season) or Epithalamin in the morning (summer and autumn). After the injection of Melatonin into adult mice the number of CFU-F and the value of the ratio CFU-F/CFU-GM decreased in false-operated animals, whereas the same indicators increased in thymusectomized animals. Under the influence of Melatonin, the ratio of CD4+/CD8+ cells increased in mice with an intact thymus and reduced in mice with a removed gland by increasing the number of CD8+ cells. The injection of Epithalamin led to the increasing in the number of CFU-F in summer, but did not lead to that in the autumn; the effect diminished in mice with a removed thymus. The orientation of changes of the indices under the influence of the pineal gland's factor in adult mice with an intact thymus corresponds to their seasonal fluctuations. After the injection of Melatonin into old mice CFU-F/CFU-GM ratio slightly reduced in false-operated animals and did not change in thymusectomized ones. Under the influence of melatonin the CD4+/CD8+ ratio of cells slightly increased in old mice with and without the thymus. Thus, the influence of the pineal gland' factors (Melatonin, Epithalamin) on the cellular structure of bone marrow, first of all on the cells of the microenvironment, in adult mice has an adaptive character and is realized with the participation of the thymus. In old mice, the thymus is also involved in the action of Melatonin on bone marrow, but the effect is less pronounced and is manifested mainly through changes in amount of CFU-GM.

[Thymic factors influence on melatonin-producing pineal function in mice of different age: the possible mechanisms].

The aging peculiarities of thymic factors influence on pineal gland function in CBA mice were investigated. Significant increase of melatonin serum level in 3 h and decrease in 24 h after one injection of thymic preparation "thymalin" in adult mice was shown. The activating influence of thymalin was depended on season and linked with increase of thymic serum factor (FTS, thymulin) level and decrease of corticosterone blood level. Thymic stroma supernatant of adult mice, which had high level of FTS, intensified in vitro melatonin-producing pineal function. The activating influence of thymic factors in vivo and in vitro on pineal function in old mice was not revealed. FTS content increased and cortocosterone level did not change in old mice after injection of thymalin. The mechanisms of thymic factors influence on melatonin level in adult organism and their peculiarities in aging were discussed.

[Circannual rhythms of bone marrow cell composition in animals during aging: the role of pineal factors].

It was investigated the influence of pineal gland's peptides (epithalamin, epithalon) and indols (melatonin) on the aging changes of circannual rhythms of stromal cells-precurcors (CFC-F), granulocyte-macrophage cells-precurcors (CFC-GM), CD4+, Mac-1+ and CD19+-cells amount in bone marrow of mice CBA. In old animals the rhythmical disturbances of the indices were characterized by loss of fluctuations (Mac 1+-cells), increase of CD4+-cells amplitude, displacement of seasonal acrophase (CFC-F), inversion of rhythm (CFC-GM), desynchronization. In old mice after epithalamin injections the season differences between the amount of Mac-1+-cells restored, CD4+-cells amplitude diminished, the amount of CFC-GM increased in spring and CFC-F diminished in autumn. The influence of epithalon on CFC-F and CFG-GM rhythm was in a smaller dose. The rhythms of some indices in old animals showed a pattern observed in adults. After melatonin injections to adult mice in winter the amount of CD4+-cells increased; the ratio CFC-GM and CFC-F changed because of increase of stromal fibroblasts. In old mice the indices were without changes.

Effect of epithalamin on the rhythm of immune and endocrine systems functioning in patients with chronic coronary disease.

In contrast to young people, changes in the rhythms of parameters manifested in elderly and senile chronic coronary patients in inversion (decreased T cell count in the peripheral blood, values of lymphocyte blastogenesis test with phytohemagglutinin, and phagocytic index in the fall), monotony (B cell count in the peripheral blood), decreased amplitude of serum hydrocortisone, and desynchronization. The levels of thymic serum factor and IgG were higher in the fall than in the spring, but the seasonal difference leveled after 2.5 years of observation. On the other hand, the rhythms of changes in the thymic serum factor and hydrocortisone were retained in patients with chronic coronary disease after 6 courses of epithalamin by the optimal protocol (period of observation 30 months) and blood T cell count increased in the fall. Improvement of the rhythms of the parameters was associated with a benign clinical course of the disease.

[Biologically active thymic factors and melatonin-producing pineal function in mice in aging].

The aging peculiarities of thymic factors influence on pineal gland function in CBA mice were investigated. It was shown that in serum of the adult mice (3-4 m) melatonin level increased significantly in 3 h after one injection of thymic preparation "thymaline". The activating influence ofthymaline depended on season and linked with increased level of thymic serum factor (FTS). Thymic stroma supernatant of adult mice with high FTS level increased melatonin-producing pineal function in vitro. There was no activating influence of thymic factors on pineal function of old mice (23-24 mo) in vivo and in vitro as well as the pineal seasonal reaction on the administration of thymaline. The importance of thymic factors influence on melatonine level for the immune system rhythmic functioning was discussed.

Antigen-induced changes in the endocrine function of the thymus in CBA mice during aging: role of peptide factors released by the pineal gland.

The titer of thymic serum factor was measured in adult and old CBA females 15, 30, and 60 min, 24, 48, 72, and 96 h after injection of 3% suspension of sheep erythrocytes and changes of this parameter under the effect of epithalamin were studied in old immunized mice. The titer of thymic serum factor increased appreciably in adult mice virtually at all periods of the study after immunization (a drop was observed only 72 h after immunization). In old mice the titer of thymic serum factor virtually did not change after injection of sheep erythrocytes, while immunization of old mice preinjected with epithalamin significantly increased this parameter. Not only the values, but their dynamics in old mice injected with epithalamin corresponded to those in immunized adult animals.

[Age-related characteristics of the melatonin-producing epiphyseal function in CBA mice immunized by T-dependent antigen].

It was investigated both melatonin blood level in the adult and old CBA-mice after immunization by sheep erythrocytes and the link between thymus and adrenal glands function in the old immunizated mice with high melatonin content in the pineal gland. It has been revealed that in the adult mice blood melatonin level was influenced by the phase changes in dynamic of immunization, namely raised significantly in 20 min with following suppression after 3 hrs and rehabilitation up to normal in 4 days. In the old mice fluctuations of melatonin level were impaired and characterized by gradually decrease in dynamics of immunization. The increase of melatonin content in pineal gland by administration of epithalamine improved the dynamics of fluctuations of thymic hormone and corticosterone level in the immunizated old mice.

[Age-related characteristics of the thymus and adrenal cortex function in CBA mice immunized by T-dependent antigen].

The thymic serum factor (FTS) titer and corticosterone level in blood of adult and old CBA mice after immunization by T-dependent antigen was investigated. It has been revealed that in adult mice these indices influenced by phase changes in dynamics of immunization whereas in old mice fluctuations of FFS and corticosterone levels were monotonous. In both adult and old mice the correlation exists between thymic hormone and corticosterone. The increasing of thymic function by thymaline administration did not improve the dynamics of FTS titer and corticosterone level in immunized old mice.

[The pineal gland's peptides factors and the rhythms of functions of the thymus and bone marrow in animals during aging]. / Peptidnye faktory épifiza i ritmy funktsii timusa i kostnogo mozga u zhivotnykh pri starenii.

It was investigated the influence of pineal peptides on the aging changes both circadian rhythm of thymic serum factor (FTS) titer in male rats Wistar and circannual rhythms of FTS, the amount of stromal cells-precurcors (CFC-F), granulocyte-macrophage cells-precurcors (CFC-GM), CD4+, Mac-1+ and CD19+-cells in bone marrow of male mice CBA. Epithalamin was injected chronically beginning from 6 month in rats and 4 month in mice. In old animals the rhythmical disturbancers of the indices characterized by loss of fluctuations, displacement of seasonal acrophase, increased or diminished amplitude, inversion of rhythm and desynchronization. After epithalamin injections in rats of 11, 17, 27 months the FTS titer at night increased and in old mice of 23-24 months appeared the peak of hormone level in summer-autumn, restored the difference between the amount of CD4+, Mac-1+-cells in bone marrow in spring and autumn, increased CFC-GM amount in spring, diminished CFC-F in autumn and increased this indice in winter. Epithalon also diminished of CFC-GM and CFC-F amount in old mice in autumn. The retarding age-related disturbances of suprachiasmaticus nucleus of hypothalamus structure, the diminishing corticosterone and testosterone levels at night, the increasing hormones level in summer and the falling in winter plays role in the improvement of thymus and bone marrow rhythmical function in old animals, reseaved epithalamin. The rhythms of most indices rhythms in old animals showed a pattern of adult.

Effect of epithalamin on circadian relationship between the endocrine function of the thymus and melatonin-producing function of the pineal gland in elderly people.

The effect of epithalamin on circadian rhythms of thymic serum factor titers and melatonin concentrations in the blood of elderly people was studied. Course treatment with epithalamin modulated the rhythm of the thymic endocrine function. The increase in the titer of thymic serum factor at night coincided with the increase in blood melatonin concentration and shift of its acrophase to 3.00, which was characteristic of young people. In elderly people with preserved nocturnal peak of the thymic serum factor titer before therapy the nocturnal (3.00) concentration of melatonin was high and did not change after drug therapy. The correlation between melatonin concentration and titer of thymic serum factor increased after epithalamin treatment.

Effects of bioactive factors of the pineal gland on thymus function and cell composition of the bone marrow and spleen in mice of different age.

The effects of factors from the pineal gland on the titer of thymic serum factor in the supernatant of 3-h thymus stroma cultures, number of stromal precursor fibroblasts and CD4+ cells in the bone marrow, and CD8+ cells in the spleens of adult and old CBA mice were studied in vitro. Epithalamin, Epithalon, and melatonin appreciably increased the titer of thymic serum factor in the supernatant of thymus stroma cultures from mice of different age and increased the percentage of CD4+ cells in the bone marrow suspension from old animals in vitro. The percentage of CD8+ lymphocytes decreased after incubation of splenic cells from old mice with melatonin. The percentage of bone marrow fibroblast precursor cells from adult and old mice did not appreciably change after incubation with the preparations.

[Circadian rhythms of pineal gland melatonin-forming function in healthy elderly people]. / Sutochnye ritmy melatoninobrazuiushchei funktsii épifiza u liudei pozhilogo vozrasta.

Circadian rhythms of pineal gland melatonin-forming function were studied in healthy elderly people (n=31, age range 60-74 years). The blood plasma melatonin concentration was determined by means of radioimmune method at 3.00, 9.00, 15.00 and 21.00. The study revealed two types of circadian rhythms of plasma melatonin concentration: a high-amplitude (29%) and a low-amplitude (71%) ones. The high-amplitude rhythm of melatonin-forming pineal gland function was observed in elderly people with a higher physical working ability and a lower functional age of cardiovascular system. It is advisable to conduct the correction of pineal gland melatonin-forming function in elderly people with low-amplitude circadian rhythm of plasma melatonin concentration.

[Regulating effect of pineal gland peptides on development of T-lymphocytes in CBA aging mice: role of microenvironment of immune system organs and neuroendocrine factors]. / Reguliruiushchee vliianie peptidov épifiza na razvitie T-limfotsitov u myshei linii CBA pri starenii: rol' mikrookruzheniia organov immunnoi sistemy i neiroéndokrinnykh faktorov.

Studies were undertaken on the development of T-lymphocytes in adult and old CBA mice and its changes at aging after injections of pineal gland peptides. It was shown that in old mice the disturbances of T-cells differentiation are registered in bone marrow, thymus, spleen and characterized by the changes of lymphocyte markers expression, migration and proliferation of cells. In old mice FTS titer, melatonin and testosterone levels decreased, the balance of noradrenalin and serotonin in hypothalamus and the cell composition of microenvironment immune systems organs impaired. After chronic (18 mo) administration of the pineal gland preparation epithalamin the amount of stromal cells-precursors, CD4+ and Mac-1(+)-cells in old bone marrow increased, improved the migration of T-cell precursors from bone marrow to thymus and their proliferative potential. The proportion of CD3+, CD4+CD8-, CD4-CD8+, Mac-1(+)-cells in old thymus increased, while that of CD44(+)-cells decreased. The proportion of CD4-CD8(+)-cells in spleen increased. The most number of indices and their balance showed a pattern of adult mice. In old mice after epithalamin the balance of amines in hypothalamus improved, concentration of melatonin in pineal gland, testosterone and FTS titer in blood increased. Epithalon has also the possibility to increase of thymic endocrine function.

[Age-related effects in thymocytes differentiation: role of pineal gland peptide factors]. / Dyferentsiiuvannia tymotsytiv pry starinni: rol' peptydnykh faktoriv epifiza.

The peculiarities of the differentiation of the thymocytes in adult and old CBA/Ca mice were studied with a help of the monoclonal antibodies to different lymphoid antigens of mice. In the thymus of old mice, the portions of the CD4-CD8-, CD44+ and CD44+Thy-1(-)-cells have been shown to be increased. The percentage of the CD4+CD8(+)-thymocytes did not change, whereas the portion of CD25+, PNA+, CD3+, CD3+CD8+, CD4+CD8- and CD4-CD8(+)-cells diminished. The ratios of different subpopulations of thymocytes, characterizing different degree of their maturation are changing with age towards the prevalence of less mature cells. After prolonged (for 18 months) administration of epithalamine the portion of CD44+, CD44+Thy-1- and CD3- 8(+)-cells in the old thymus has been shown to be decreased, while that of CD25+, CD3+, CD3+CD8+, CD4+CD8/and CD4-CD8(+)-cells increased. According to many parametres thymus of those old animals corresponds to adult mice. Epithalamine raised the titer of one of the thymic hormones--thymic serum factor, both in the blood of the old mice and in the supernatant of a 3-hour culture of the thymic stroma.

[Age-related changes in circadian and circannual fluctuations of the immune response and the number of cells in lymphoid organs of animals: a possible connection to thymic factors]. / Vikovi zminy tsyrkadnykh i tsyrkanual'nykh kolyvan' velychyny imunnoï vidpovidi ta chysla klityn u limfoïdnykh orhanakh tvaryn: mozhlyvyi zv'iazok z faktoramy tymusa.

The 24-hour and seasonal fluctuations of cellularity of thymus and spleen and antibody levels in male CBA/Ca mice and Wistar rats of different ages were investigated. In adult rats (6-11 mo) the peaks of thymus and spleen cellularity were at nights and in the daytime, respectively. In young mice (3 mo) the thymus and spleen cellularity were highest in autumn and spring, respectively. The titer (log2) of hemolysines in autumn was 16 times greater than in spring. Correlations between the rhythmical changes of cellularity of thymus and spleen and between the latter and the factor thymic serique (FTS) titer were found. With age (4 month and onward in mice and 17 months in rats), the peaks of amounts of cells in thymus and of its mass in night and in autumn and of the antibodies titers in autumn were diminished. The displacement of the acrophases of indicers to other seasons was seen. In old animals the rhythm of indices became monotonous. In aging the intrasystemic relations underwent changes (new relation appeared, while the existing ones weakened or disappeared). The chronic (18 mo) administration of thymus preparation thymalin increased FTS titer and promoted the appearance of the peak of antibodies in autumn.