[Prion diseases and the biosecurity problems.]

The review presents the current state of the problem of prions and prion diseases with an emphasis on theepidemiological and epizootological risks of pathogens that cause fatal neurodegenerative diseases in humans and animals. The results of molecular genetic studies of the conversion of normal PrPc prion protein molecules to infectious forms of PrPd, resistance to physical disinfection methods, in particular exceptional thermal stability, and their ability to overcome interspecific barriers, while increasing virulence, are described. The possibility of infection not only by nutrition, when eating even heat-treated meat of sick animals, but also due to surgical interventions, especially neurosurgical and ophthalmic, as well as the use of immunobiological preparations, are emphasized. Since there are currently no means for the effective treatment of prion diseases in the world, attention is drawn to the high degree of relevance for the biosafety of the country to develop domestic highly sensitive test systems that can effectively detect prion infectious protein in vivo at the preclinical stage of the disease. The latest methods of automatic protein amplification and identification of prion proteins are briefly described as the most promising areas of research in the field of diagnosis of prion diseases.

Skin Fibroblasts as the Object for Clinical Diagnosis of Parkinson's Disease in Persons of Different Ages.

We compared the expression of Aß42 peptide, τ-protein, and α-synuclein in the substantia nigra and skin fibroblasts of elderly and senile patients with Parkinson's disease and subjects without neuropathology. Expression of markers in the studied tissues was assessed by immunohistochemical and immunocytochemical methods. The expression of Aß42 peptide, τ-protein, and α-synuclein in the substantia nigra of elderly and senile patients with Parkinson's disease was higher by 11-31 times than in subjects without neuropathology. In skin fibroblasts of patients with Parkinson's disease, the expression of Aß42 peptide and α-synuclein was 3-14 times higher than in subjects without neuropathology, and expression of τ-protein did not significantly differ in the studied groups. Thus, immunocytochemical analysis of the expression Aß42 peptide and α-synuclein in skin fibroblasts can be a simple method of early diagnosis of Parkinson's disease in elderly persons.

Expression of Aß42, τ-Protein, p16, p53 in Buccal Epithelium: Prospects for Use in the Diagnostics of Alzheimer's Disease and Rate of Aging.

The expression of Aß42, and τ-protein, and p16 and p53 proteins was analyzed in the buccal epithelium of elderly and senile patients with Alzheimer's disease. We revealed enhanced synthesis of Alzheimer's disease markers Aß42 (by 15-30 times) and τ-protein (by 5 times) in comparison with the corresponding values in people without neurodegenerative pathology of the same age groups. In addition, increased synthesis of proteins of cell aging and apoptosis p16 (by 6-10 times) and p53 (by 2-3 times) was observed in patients in comparison with age-matched persons without neuropathology. These data suggest that complex analysis of the expression of Aß42, τ-protein, p16, and p53 in the buccal epithelium is a promising method for in vivo diagnosis of Alzheimer's disease and assessment of the rate of aging during the development of this pathology.

[Melatonin as a molecular marker of age-related pathologies].

The review has described melatonin as a prognostic marker of invasive and non-invasive diagnostic of organism aging time and age-related pathology. Decreasing of melatonin level in buccal cells has correlated with patient age. Melatonin level in patients with Alzheimer disease has decreased. Melatonin level in blood plasma has correlated with severity of menopausal syndrome. Melatonin secretion in enterocytes increased during gastric ulcer. In oncology patients was described changes of 6-COMT - metabolite of melatonin in urine in dependent of histology type and stage of disease. Thus, melatonin is the molecular marker, which characterized integral processes in neuro-immuno-endocrine system and can be verified by non-invasive methods in peripheral tissues and biological fluids of organism.

[Molecular markers of Alzheimer disease early diagnostic: investigation perspectives of peripheral tissues.]

Alzheimer's disease (AD) is a progressive neurodegenerative disorder of elderly and old age people. For intravital diagnosis of the expression of signaling molecules - AD markers, cerebrospinal fluid (CSF) and peripheral tissues are used: lymphocytes and blood platelets, buccal and olfactory epithelium, skin fibroblasts. There are several changes in the production of hyper phosphorylated form of τ-protein, BACE1 and peptide Аß42 in CSF in case of AD, but CSF taking may have a number of side effects. Less traumatic taking of sampling tissues for the diagnosis of AD is in use of epithelium biopsy and blood portion. An increase in the expression of the hyper phosphorylated form of τ-protein is shown in blood lymphocytes of AD patients. An increase in the content of high molecular weight forms of phosphorylated t-protein and amyloid precursor protein-APP was also revealed in blood platelets of AD patients. Changes in the amount of 2 miRNA families - miR-132 family and miR-134 family were revealed in blood cells 1-5 years before the manifestation of clinical signs of AD. An increase in the concentration of bound calcium, synthesis of peptides Aß40 and Aß42, τ protein was observed in AD skin fibroblasts. In the olfactory and buccal epithelium an increase in the expression of hyper phosphorylated form of τ-protein and Aß peptide was detected in patients with AD. Verification of AD markers in peripheral tissues for biopsy have the important significant for life diagnostics, prevention and and target AD treatment.

[Slow infections of humans and animals].

This review is dedicated to the 60-th anniversary of the exploration of slow infections of humans and animals caused by viruses and prions.

[Life extension study on high-yield non-myeloablating bone marrow transplantation from young to old mice].

Tissue renewal is the known phenomenon, when the progeny of resident or circulated stem cells (SC) replaces the vanishing cells. The delivery of stem cells via circulation should result in stem cell homing and differentiation into wide variety of tissues and gives promise as therapy for many diseases of tissue failure including aging itself. To test this hypothesis, we created chimeric mice C57BL/6 by bone marrow (BM) transplantation from young 1.5 months old donors to 21.5-months old recipient mice of the same strain C57BL/6. We applied here the recently published new transplantation technique, which allows to get high scores of chimerism due to very high amount of transplanted cells (1.5 x 10(8) per mouse or 25% of its total BM count). In the earlier works only 1% of total BM count (about 5 x 10(6) cells per mouse) was usually transplanted to lethally irradiated mice, what excluded the possibility to apply this method for life extension. As a result of the modified technique implementation, the mean post-transplantation life (starting the 21.5 months old) of treated mice was 4.9 months versus 3.4 months for untreated mice. The difference in 1.5 months counts for 44% extension of mean post-transplantation life.

[Chronic persistent infections of anterior segment of eye: clinico-laboratory aspects].

AIM: Improvement of therapy of chronic ophthalmic infectious diseases during assessment of functioning of different arms of immune system. MATERIALS AND METHODS: Three hundred and fifty patients with chronic red-eye syndrome were tested by immunofluorescence assay on the presence of antigens of herpesviruses, adenoviruses and Chlamydia in samples from conjunctiva. Expression of 11 cytokines' genes was measured in peripheral blood mononuclear cells by reverse transcription polymerase chain reaction. Production of IFN-alpha and IFN-gamma, levels of serum and spontaneously produced interferon as well as level of susceptibility to the range of immunomodulating preparations were measured during study of interferon status in whole blood cells. Study of parameters of cytokine, interferon and immune statuses was performed in 70 patients. Counts of T- and B-lymphocytes, T-helpers, NK-cells as well as level of circulating immune complexes were measured during study of immune status. RESULTS: Antigens of herpes simplex virus and adenoviruses were detected in samples from conjunctiva in 27% (95 persons) and 36% (126 persons) of patients respectively. Enhanced level of expression of several cytokines (IL-2, IL-4) in studied patients compared with healthy volunteers was observed. Expression levels of IL-12 and TNF-alpha mRNAs were, in opposite, in 2 - 3 times lower. Disorder of IFN-alpha and IFN-gamma synthesis on post-transciption level was observed in 60 - 90% of patients. Decrease of absolute numbers of total T-lymphocytes and T-helpers as well as increase of absolute number of NK-cells was noted in 20%, 25%, and 27% of patients respectively. CONCLUSION: Assignment of individually oriented antiviral, antibacterial and immunomodulating therapy allowed to mitigate intensity of clinical symptoms in 30 -60% of patients with chronic persistent infections of anterior segment of eye.

Transfer of signs of aging to young mice by splenic lymphoid cells from old syngeneic donors.

Single intraperitoneal injection of splenic lymphoid cells from 20-month-old mice to 2-month-old syngeneic mice (similarly as 5-day injections of purified brain extract) leads to preterm (4 months earlier than in the control) appearance of aging factor in the blood (the main sign of old age). Combined injections of brain extract and splenic lymphoid cells led to the appearance of aging factor in the blood 5 months earlier than in the control.

Artificial aging of mice.

Clinical signs of aging verified by morphometrical analysis of brain tissue were observed in young mice 4 months after administration of brain extract from old mice (5 intraperitoneal injections).

[Gliosis as a trigger of pathomorphological changes in prion diseases]. / Glioz kak puskovoi mekhanizm patomorfologicheskikh izmenenii pri prionnykh bolezniakh.

Experiments with three cell lines revealed that the scraplecontaining cerebral extract, obtained from preliminarily infected 6-month mice, sharply induced the cellular proliferation, which was registered yet in 3 days after incubation. However, the cerebral extract of healthy 6-month mice did not virtually influence the velocity of cells' reproduction in all three cultures. The authors suggest, with respect to published data and to their independently found research results, that the gliosis of primary importance in shaping up the pathomorphological alterations in the cerebral tissue in prion diseases of man and animal.

[From prion diseases to the problem of aging and death]. / Ot prionnykh boleznei k probleme stareniia i smerti.

The brain tissue of aging mice shows a factor that stimulates the proliferation of cells of glial origin both in primarily trypsinized and inoculated cultures. This factor that is likely to be of peptide origin is characterized by pronounced accumulation dynamics with increasing age in mammals, by unusually high thermal stability and by its possible detection in the brain extracts by means of isoelectrofocusing rather than electrophoresis. The author proposes a concept under which the brain tissue of aging mammals, including human beings, accumulates the factor that is active in stimulating the proliferation of glial elements, thus resulting in impairment of neuronal trophism, which can in turn be a cause of their death and, in the long run, a cause of death of the brain and the whole body.

[The immunological consequences of congenital infections]. / Immunologicheskie posledstviia vrozhdennykh infekstii.

Mice of different strains were inoculated with type A influenza virus or Mycoplasma arthritidis in the second half of pregnancy. A part of the animals born after this inoculation were characterized by a sharp retardation of growth. The study of the immune status of such animals revealed that their proliferative response to mitogenic/superantigenic factors of the infective agents introduced during pregnancy was suppressed or absent, and the cells of their immune system began to recognize syngeneic intact stimulators in the mixed lymphocytes culture as heterogeneous ones. The spleen of the experimental animals was found to contain suppressor cells, both specific and nonspecific with respect to the infective agent. After inoculation with M. arthritidis areactivity was observed only in mice, sensitive to mycoplasmal superantigen. The data thus obtained suggest that the penetration of infecting agents producing mitogenic/superantigenic factors induced changes in the immune system, contributing to the persistence of the infective agent in the host body.

The Possible Immunological Mechanisms of Pathology in Mice with Congenitally Acquired Influenza Infection.

The fact that congenitally acquired viral infection often strongly influences specific and non-specific immunoreactivity is well documented. Viral infection of pregnant female may lead to serious of pathological consequences for the offspring, namely, to mortality, developmental disorders and in less severe cases to body growth retardation, wasting syndrome and immunodeficiency. In this connection, we have studied congenitally acquired influenza infection in CII mice. The progeny of C57BL/6 female mice, which were infected with influenza virus (A/WSN) by the 3rd week of pregnancy, exhibited a profound growth retardation and major morphological lesions of central nervous system, lymphoid and other organs. We have found out that mice with congenitally acquired influenza infection had autoreactive killer T cells in their lymphoid organs. CII mice exhibited some features of chronic immune activation, namely elevated spontaneous proliferation, spontaneous development of plaque forming cells, and spontaneous inhibition of migration activity. Lymphoid cells from mice with congenitally acquired influenza infection induced an enlargement of regional lymph nodes after they had been injected into syngeneic non-infected recipient in popleteal node assay. The level of this reaction depended on the level of virus-bearing cells in donor cell population and correlated with the increase of gammadelta and CD4(+) T cells. The role of these interactions in pathology is discussed herein.

[The induction by the influenza virus A/H1N1 (serovariant Hsw1N1) and its glycoproteins of congenital pathology in mice]. / Induktsiia virusom grippa A/H1N1 *serovariant Hsw1N1) i ego glikoproteidami vrozhdennoi patologii u myshei.

Influenza A/H1N1 (serovariant Hsw1N1) virus, a sum of isolated glycoproteins, separately neuraminidase "heads", inoculated into white random-bred female mice, induced in some of the offsprings the pathology clinically and pathomorphologically similar to previously described slow virus infection. At the same time, the pathology in the offsprings caused by the antigenic virus variant under study was characterized by complete absence of fur and more dynamic progress of the disease. It is quite obvious that glycoproteins, particularly neuraminidase, are the molecular biological basis of dystrophic and degenerative changes in the organs of baby mice due to desialization and increased fluidity of capillary endothelium membranes and, possibly, CNS and other organ cells.