[Effective Viral Delivery of Genetic Constructs to Neuronal Culture for Modeling and Gene Therapy of GNAO1 Encephalopathy].

GNAO1 encephalopathy is an orphan genetic disease associated with early infantile epilepsy, impaired motor control, and severe developmental delay. The disorder is caused by mutations in the GNAO1 gene, leading to dysfunction of the encoded protein Gao1. There is no cure for this disease, and symptomatic therapy is ineffective. Phenotypic heterogeneity highlights the need for a personalized approach for treating patients with a specific clinical variant of GNAO1 and requires the study of the disease mechanism in animal and cell models. Towards this aim, we developed an approach for modeling GNAO1 encephalopathy and testing gene therapy drugs in primary neurons derived from healthy mice. We optimized the delivery of transgenes to Gαo1-expressing neurons using recombinant adeno-associated viruses (rAAV). We assessed the tropism of five neurotropic AAV serotypes (1, 2, 6, 9, DJ) for Gαo1-positive neurons from the whole mouse brain. The DJ serotype showed the highest potential as a reporter delivery vehicle, infecting up to 66% of Gαo1-expressing cells without overt cytotoxicity. We demonstrated that AAV-DJ also provides efficient delivery and expression of genetic constructs encoding normal and mutant Gαo1, as well as short hairpin RNA (shRNA) to suppress endogenous Gnao1 in murine neurons. Our results will further simplify the study of the pathological mechanism for clinical variants of GNAO1, as well as optimize the testing of gene therapy approaches for GNAO1 encephalopathy in cell models.

[Rudolf Virchow's life path of as a reflection of the development of medicine in the 19th century (on the occasion of 200th birth anniversary)]. / Zhiznennyi put' Rudol'fa Virkhova kak otrazhenie razvitiya meditsiny XIX veka (k 200-letiyu so dnya rozhdeniya).

The article describes the life path of Rudolf Virchow (1821-1902), an outstanding German pathologist, one of the founders of scientific medicine, the creator of the scientific system known as cellular pathology, the author of numerous works on general biology, anthropology, ethnography, and archaeology. It assesses the contribution of the scientist to the development of science and culture in the nineteenth century. A relationship is shown between the formation of a modern scientific approach in medical sciences and the multifaceted activities of Rudolf Virchow.

[Alexey Ivanovich Polunin (1820-1888). On the occasion of the 200th birth anniversary]. / Aleksei Ivanovich Polunin (1820­1888). K 200-letiyu so dnya rozhdeniya.

The paper describes the life of A.I. Polunin (1820-1888), the first professor of an independent department of pathological anatomy in Russia, which was founded at Imperial Moscow University in 1849. It analyzes his doctoral dissertation on cholera and also gives the reviews of his key works. It is noted that the activities of A.I. Polunin as a scientist were greatly influenced by the humoral theory of pathology by Karl von Rokitansky (1804-1878), the doctrine of blood «crases¼, and by the works of representatives of the Viennese School of physicians who upheld the clinical and anatomical approach in medicine. At the same time, Professor A.I. Polunin is noted to be the first to familiarize Russian physicians with the theory of cellular pathology by R. Virchow (1821-1902).

[Syndrome of bacterial overgrowth in patients with the reduced stomach acid secretion: some aspects of the diagnosis].

OBJECTIVE: To study the frequency of occurrence of small intestinal bacterial overgrowth (SIBO) in GERD patients with reduced stomach acid secretion receiving long-term PPI treatment, and patients with chronic atrophic gastritis with reduced stomach acid secretion, using the hydrogen breath test (HBT) and studies of short-chain fatty acids (SCFA) in various biological substrates; compare the obtained results. MATERIAL AND METHODS: There were 100 people surveyed: 1 group consisted of 40 patients with chronic atrophic gastritis (HG) with reduced stomach acid secretion; 2 group consisted of 60 patients of GERD in patients receiving PPI different duration (0-6 months, 6-12 months more than 12 months). All the patients were examined by a load of lactulose and research SCFA using gas-liquid chromatographic analysis (GC-analysis) in various biological substrates (duodenal secretion and feces). RESULTS: When performing HBT in patients with chronic atrophic gastritis frequency detection SIBO amounted to 57.5 per cent. In GERD patients receiving PPI SIBO was detected in 8.3% of cases (0-6 months of treatment), 21.7% (6-12 months of treatment), 61.6% (over 12 months). 15% of HG patients and 13.3% of GERD patients receiving PPI, with clinical manifestations of SIBO, the data turned out to be negative. The SCFA parameters in the duodenal secretion in patients with GERD during PPI therapy depending on the duration of the admission and in HG patients with reduced stomach acid secretion were studied, as well as in patients with clinical manifestations of SIBO depending on the data of HBT (+/-). We also studied SCFA parameters in patients with positive HBT results depending on time of registration of the increase of hydrogen concentration. It is established that the study of SCFA in the duodenal secretion is an important diagnostic SIBO test, which allows not only to detect increased activity of the microflora, but also to determine its tribal affiliation. In some cases, its information value exceeds the HBT. The study of SCFA in various biological substrates (duodenal secretion, feces) allows you to specify the localization of existing violations. The rapidity of obtaining results allows to consider this method as screening.

[Use of Pervomur for prophylaxis of infectious complications in elective surgeries of abdominal organs]. / Primenenie pevromura dlia profilaktiki infektsionnykh oslozhnenii pri planovykh operatsiiakh na organakh briushnoi polosti.

After the experimental investigations performed, the 0.6-0.8% solution of Pervomur (as irrigation of the operation area or application) was used in 103 patients. As compared with the control group the intraoperative using of Pervomur allowed the frequency of postoperative complications to be reduced from 21% to 15% and the time of treatment was shortened.

Education programme on aerospace and environmental medicine for medical faculty of Lomonosov Moscow State University.

The problems and approaches to organisation of the education process in the field of aerospace and environmental medicine for medical students are discussed. Original education developed on the basis of Russian experience in space biology and physiology, environmental medicine, aerospace medicine and medical support during spaceflight. The main goals of these programs are to acquaint students with: interaction of living organisms with natural and artificial surroundings, including space flight conditions; the physiological reactions on extreme environmental factors; basic mechanisms of human adaptation to space flight and particularly to microgravity; the current research in space medicine and new telecommunication technologies. All programs are formed in accordance with contemporary progress in life sciences and revealed a result of the interdisciplinary approach to education process.

[Several indicators of tissue oxygen during modeling of extravehicular activity of man]. / Nekotorye pokazateli kislorodnogo rezhima tkanei pri modelirovanii vnekorabel'noi deiatel'nosti cheloveka.

Investigations of tissue oxygen indices during simulation of extravehicular activity (EVA) of cosmonauts demonstrated that breathing pure oxygen at approximately 280 mmHg elevates oxygen tension in capillary blood, and capillary-tissue gradient during physical work. Physical work alone stimulates tissue oxygenation due to, apparently, intensification of the processes of oxidative phosphorylation. The observed shifts in oxygen status reverse significantly within the first 5 min after completion of the experiment.

[Thermodynamic effectiveness of active transport of calcium ions]. / Termodinamicheskaia éffektivnost' aktivnogo transporta ionov kal'tsiia.

Computation of variation of general and partial free energy, exemplified by intracellular transport of calcium ions performed by sarcoplasmic reticulum Ca(2+)-ATPase, is used to demonstrate that the system of active transport of calcium ions works effectively even if transmembrane difference of electric potentials is absent. The possibility that local redistribution of charges on molecules complexes influences the Ca2+ transfer, is discussed.

[Binding of phenyldicarbaundecaborane with proteins, membrane fragments, and whole cells]. / Sviazyvanie fenildikarbaundekaborana belkami, fragmentami membran i tselymi kletkami.

The binding of the lipophilic synthetic anion, phenyldicarbaundecaboran, to inner and outer membranes of myocytes, platelets and erythrocytes as well as to integral and surface membrane proteins has been studied. It has been found that the distribution of lipophilic synthetic ions in the system biopreparations--aqueous phase is not determined by the value of the transmembrane difference of electric potentials but, rather, by ion adsorption by the protein-lipid components of the cell. The latter is controlled by the correlation between the volume of hydrophobic areas and the local electrochemical characteristics of the cell.

[Structural and functional disorders of erythrocyte membranes in simulated extravehicle space work]. / Strukturno-funktsionalnye narusheniia membran éritrotsitov pri modelirovanii vnekorabelnoi deiatelnosti cheloveka.

The experimental data on a structural-and-functional status of erythrocyte membranes in ground-based simulation of human extravehicular activity (EVA) are discussed. The changes in the structure and function of erythrocyte membrane were determined from the fixation of lipophilic ions, specific activity of ion-transporting adenosine triphosphatases and content of the products of peroxide oxidation of lipids (POL) in plasma. On exposure to hypobaric hyperoxia during real EVA the membrane status changes are associated with the activation of a cascade of POL processes.

[The permeability of rat erythrocyte membranes for sodium and potassium ions during exposure to pulsed electromagnetic field under head-down-tilt hypokinesia]. / Pronitsaemost' membran éritrotsitov krysy dlia ionov natriia i kaliia pri vozdeistvii impul'snogo élektromagnitnogo polia v usloviiakh antiortostaticheskoi gipokinezii.

The effect of a low-frequency (1 and 10 Hz) pulse electromagnetic field (PEMF) on permeability of erythrocyte membrane in 47 rats while being exposed to a 14-day head-down tilt (HDT) and in 60 control rats was studied. After exposure to PEMF (20 min, 6-24 mT) the rats were Na(+)-, K(+)-cotransport and Na+,Na(+)-metabolism, Na and K losses as well as specific activity of Na(+)-, K(+)-ATP in the erythrocytes have been measured. An ability of PEMF to inhibit an ATP activity by 20-30% and to increase the rate of ions loss from the cells has been revealed. Under HDT, these effects are preserved. The possible mechanisms of PEMF effect on an ionic permeability of the membranes are discussed.

[Oxygen tension and water-soluble products of lipid peroxidation in blood of volunteers in hypobaric hyperoxia]. / Napriazhenie kisloroda i vodorastvorimye produkty perekisnogo okisleniia lipidov v krovi ispytuemykh pri gipobaricheskoi giperoksii.

Oxygen tension in the arterial blood with transcutaneous method (tcPO2) and TBA-active products of venous blood plasma were measured during simulation of extravehicular activity. There was a parallel increase of tcPO2 and the level of TBA-active products upon introduction of hypobaric hyperoxia factor. Detrimental action of lipid peroxidation products on erythrocyte membranes can be one of the factors of anemia during space flight.

[Accumulation of calcium ions in myocardial sarcoplasmic reticulum of restrained rats exposed to the pulsed electromagnetic field]. / Akkumuliatsiia ionov kaltsiia sarkoplazmaticheskim retikulumom serdechnoi myshtsy krys v usloviiakh ogranichennoi podvizhnosti i pri vozdeistvii impulsnym elektromagnitnym polem.

Alteration in the velocity of Ca++ accumulation was investigated in myocardial homogenates from restrained rats which were chronically (1 and 2 months) exposed to the impulse electromagnetic field with frequency 1 Hz and magnetic induction 6-24 mT. The fall of the studied variable by 33% was observed in a month while restoration of the velocity of Ca++ transport was shown after two months of exposure. The results are discussed in terms of the organism adaptation to physico-chemical and physiological impacts.

The evaluation of biological efficiency of electromagnetic fields generated by implanted radiotelemetric transmitters used in space research on animals.

The study was carried out in 50 male rats abdominally implanted with biotelemetric systems (BTS) or mock-up. The animals were provided with 12/12 light/dark schedule during 6-week experiment. The electromagnetic field (EMF) frequency was 455 kHz, magnetic induction near transducer was about 10(-2) mT. Circadian rhythm of the body temperature and locomotor activity was controlled in course of the experiment. The latter been finished, some tissues and the blood of the animals have been sampled to test corticosterone, testosterone, T3, T4 level in serum with radioimmunoassay and membrane permeability for Ca2+, Na+, K(+)-ATPase activity and charge changes in liver microsomes was detected. The probable ways of the EMF influence on whole body are discussed.