[Nanosystems and targeted transport of medicinal preparations to the brain].

Intense efforts in the last decades have resulted in numerous inventions to improve drug delivery to CNS. Some of them have significant potential for clinical applications. At the same time, the variety of these strategies is in itself indicative of great difficulties inherent in the transfer of therapeutic and imaging agents across the blood-brain barrier. Combination of several approaches, such as encapsulation of drugs in nanocontainers, incorporation in micellar nanostructures, the use of Pluronic "unimers" for inhibition of drug efflux in endothelial cells of brain capillaries, appears to be the most promising modality. The same refers to the methods of intracellular transport of drug suspensions developed recently. All these achievements provide a solid basis for the further improvement of drug pharmacokinetic properties, reduction of systemic toxicity, increase of therapeutic efficacy, and earlier diagnosis of CNS pathology.

Development of enzyme immunoassay of glial fibrillary acidic protein on the basis of recombinant antigen.

Quantitative enzyme immunoassay of glial fibrillary acidic protein of astrocyte intermediate filaments opened new prospects for highly selective diagnosis and monitoring of pathological processes in the CNS. Immunochemical screening of glial fibrillary acidic protein in biological fluids helps to adequately evaluate the permeability of the blood-brain barrier in CNS diseases associated with violation of its functions, such as hypoxic and ischemic disorders, neuroinfections, glial tumors, brain injuries, etc. Wide-scale introduction of enzyme immunoassays into clinical laboratory practice implies the development of biotechnological approaches to the creation of methods for obtaining EIA components. This paper presents a method for creation of a test system for EIA of glial fibrillary acidic protein (GFAP) on the basis of recombinant GFAP and antibodies obtained by immunization with recombinant GFAP. Due to this approach, a highly standardized test system for the analysis of GFAP in human biological fluids was created.

Cloning and expression of rat GFAP cDNA in Escherichia coli.

A cDNA fragment encoding GFAP was amplified by reverse transcription PCR from total mRNA isolated from primary culture of rat astrocytes and cloned for expression in Escherichia coli using pET-28a vector. High level of GFAP expression was confirmed by SDS-PAGE, while immunochemical identity was verified by immunoblotting. The constructed producer strain is a cheap source of GFAP and can be used for diagnostic purposes.

[The role of clinical suicidology in the system of measures directed towards lowering early mortality and increasing population life span].

The level of early mortality has increased substantially in Russia within the last fifteen years, having exceeded the same parameter in developed countries and in the entire post-Soviet area. The second frequent reason for early mortality is a group of factors that includes accidents, suicides, murders, and other external causes. The proportion of suicides in this group is 45 to 50%. As a result, in the recent years the suicide rate in Russia has filled the second place in the world. The authors of this article analyze the suicide rate in different Russian regions, distinguishing between regions with the highest and lowest rate, and characterizing population risk factors of suicidal danger, a special place among which is filled by socioeconomic condition of the regions and the ethnic composition of their population. Increase of the effectiveness of suicide prevention depends on creation of adequate scientific basis, which clinical suicidology can become. The authors substantiate the necessity to distinguish suicidology as a separate field of clinical medicine, and formulate its definition, goals and objectives. Basing on the obtained results, the authors come to a conclusion on the medico-social importance of the development of clinical suicidology in lowering early mortality and increasing the country's population life span.

[Direct drug delivery to the brain].

The authors of the review discuss the present-day state of and promising approaches to directed delivery of biological agents into the brain. Special attention is drawn to micellar and liposomal transport through the blood-brain barrier (BBB) targeted by immunochemical vectors, such as native or hydrophobized antibodies to specific antigens located at the BBB or in the brain parenchyma.

[The EEG correlates of delayed mental development in adolescents].

Fifty-eight male adolescents aged 15-17 years have been divided into 3 groups: 31 patients with mental infantilism syndrome were included in the study group, 14 with organic brain disorders--in the comparison group and 13 psychiatrically and neurologically normal subjects--in the control group. EEG was recorded from 12 leads monopolarly at rest, during hemisphere-specific cognitive tasks performance and exposure to aversive sound stimulation. Two-Hz wide EEG spectral ranges--A, theta1, theta2, alpha1 and alpha2--were analyzed. Only in the study group, there were decreased values of alpha2 spectral power (SP) and reduced reactivity to functional tests and insufficient lateral differentiation of the reactions during the cognitive tasks performance. Maximal deviation of the SP reactivity indices as compared to the comparison group was found in alpha2 range. The disturbances obtained indicate the delay of brain maturation and retardation in formation of specific neural networks. The results of the study demonstrate that indices of alpha2 SP may be considered as objective criteria of delayed mental maturation.

[Production and characteristics of PEGylated immunoliposome transport vectors specific for neural tissue astrocytes].

PEGylated (stealth) immunoliposomes covalently linked to antibodies against human gliofibrillary acidic protein (GFAP) were prepared by coupling the thiolated monoclonal anti-GFAP antibodies, D4, with a maleimide derivative of the phosphatidyl ethanolamine of the liposomal membrane. Depending on the initial protein-to-lipid ratios, the immunoliposomes prepared (with a diameter of about 70 nm) were coupled with 60 to 240 molecules of the antibodies. In vitro experiments with cultures of the embryonic rat brain astrocytes demonstrated a specific binding of these immunoliposomes, which could be inhibited by the preincubation of the cells with free non-coupled D4 but not with non-specific antibodies. Administered intravenously into rats, the immunoliposomes exhibited a kinetic behaviour typical for the PEGylated liposomes: after 24 h, approximately 20% of the doses injected were still present in the blood; the elimination rate constants were 0.05-0.09 h(-1), the elimination half-lives were 8-15 h. With such a systemic longevity, as well as with such a specificity, these immunoliposomes, non-penetrating through intact blood-brain barrier (BBB), should be useful in delivering pharmacological agents to glial brain tumours (which continue to express GFAP) or to other pathological loci in the brain with a partially disintegrated BBB.

Culturing of specialized glial cells (olfactory ensheathing cells) of human olfactory epithelium.

A monolayer of dissociated glial cells of human olfactory epithelium was cultured in Petri dishes and 12-well plates using a polylysine-laminin substrate. Primary cultures were subcultured after 10-15 days. The cell cultures were analyzed by phase contrast microscopy at all stages of culturing. A cytological study involved histological methods (trypan blue staining) and immunocytochemical visualization of GFAP, nestin, and low-affinity nerve growth factor receptors. At the final stage of culturing (5 passages) the monolayer cultures included 2 types of cells: GFAP- and p75-positive glial cells and nestin-positive fibroblasts.

[Liposome-oriented transport of therapeutic drugs]. / Napravlennyi transport lekarstvennykh sredstv s pomoshch'iu liposom.

The attempts to use liposomes as containers for the transport of therapeutic drugs have been undertaken during the recent 40 years. However, the first success was achieved only in the 80-ies, when the sterically stabilized liposomes were invented. It was found that the liposome biological layer modified through, adding to it, certain polymers prolonged the blood circulation and reduced the capture of liposomes by RES cells. Elaboration of immunoliposomes, i.e. those conjugated with antibodies, was the next step in the path of perfecting the liposomes as a transport tool for the sake of binding with target-cells and to ensure the address-oriented delivery of drugs to a pathology focus. Preclinical and clinical testing of liposome-form of antitumor drugs witnessed to their lower toxicity and better pharmacokinetic indices; besides, they selectively accumulate themselves in tumor cells and have a more pronounced therapeutic effect even at lower doses of drugs in case of tumors resistant to the already made chemotherapy.

[Modeling of focal ischemia of the brain]. / Modelirovanie fokal'noi ishemii golovnogo mozga.

Methods of modeling and criteria of evaluating the pathological process in the central nervous system (CNS) as well as modern technologies of provoking the focal ischemia of the brain in experimental animals are under discussion. The results were analyzed comparatively from the viewpoint of efficiency and adequacy of certain models as well as of the clinical specificity of ischemic strikes in man and of set research goals. A literature analysis confirms that the existing arsenal of methodical schemes provides for choosing the most adequate model of focal ischemia of the brain and to ensure a cerebral infarction of a preset scope and localization; it makes possible also an objective evaluation of pathological processes occurring in the cerebral tissues both at the earliest stages of ischemic lesion and during a relatively long time period comparable with rehabilitation time period. Achievements in the sphere of experimental modeling of focal ischemia of the brain pave the way to further promotion of experimental therapy in acute stroke and open up new research priorities; it concerns primarily research of mechanisms timing the neurodegenerative process after ischemic stroke as well as searching-for and testing of means of stroke prevention and of patients' rehabilitation.

Selective accumulation of monoclonal antibodies against neurospecific enolase in brain tissue of rats with middle cerebral artery occlusion.

Preparations of I(125)-labeled monoclonal antibodies against neurospecific enolase and mouse plasma IgG1 were injected intravenously to rats immediately after unilateral occlusion of the middle cerebral artery. Radioactivity of I(125)-labeled monoclonal antibodies against neurospecific enolase in the brain tissue progressively increased, reached a maximum by the 48th hour, and remained practically unchanged after 72 h. At the same time radioactivity of labeled IgG1 in the brain tissue and radioactivity of both preparations in the blood, liver, spleen, kidneys, heart, and lungs decreased over 72 h. Selective accumulation of I(125)-labeled monoclonal antibodies against neurospecific enolase was less significant in the brain tissue of the contralateral hemisphere and cerebellum not exposed to ischemia.

Enzyme immunoassay of NSE and GFAP as the criterion of dynamic evaluation of the rat blood-brain barrier in perinatal hypoxic ischemic injury of the CNS.

Enzyme immunoassay of the serum neurospecific antigens (gliofibrillar acid protein and neurospecific enolase) was used for evaluation of the resistance of the blood-brain barrier in Wistar rats with perinatal hypoxia and ischemia of the CNS. Perinatal hypoxia and ischemia of the CNS was modeled by two methods: ligation of the common carotid artery in 7-day-old rats followed by 3.5-h hypoxic hypoxia or 15-min anoxic exposure of fetuses isolated via hysterectomy on day 21 of gestation. Enzyme immunoassay of serum gliofibrillar acid protein and neurospecific enolase in control an experimental rat pups was carried out once a week during 3 months. In controls serum levels of gliofibrillar acid protein and neurospecific enolase virtually did not change during postnatal development, while in animals with cerebral hypoxia and ischemia induced in fetuses by both methods serum concentration of neurospecific enolase sharply increased 1 week after the injury and increased on weeks 6 and 10. The content of gliofibrillar acid protein was maximum on week 1 and later considerably varied, the peaks of its concentrations observed on weeks 3 and 8 preceded the increase in neurospecific enolase activity in peripheral blood.

[Myelin oligodendrogliocyte glycoprotein: the structure, functions, role in pathogenesis of demyelinating disorders]. / Mielin-oligodendrogliotsitarnyi glikoprotein: Stroenie. Funktsii. Rol' v patogeneze demieliniziruiushchikh rasstroistv.

Myelin oligodendrocyte glycoprotein (MOG) is a myelin-specific protein of the central nervous system (CNS). It is a member of the immunoglobulin superfamily. The contents of this protein in oligodendrogliocyte membrane is very low (approximately 0.1% total proteins). Functions of MOG are still unknown. It is considered that MOG is an autoantigen capable to produce a demyelinating multiple sclerosis-like disease in experimental animals. The structure, membrane topology, putative functions and role in demyelinating diseases are considered in this review.