A Method for the Production of Recombinant VSVs with Confirmation of Biological Activity.

The design of new effective cancer treatment methods is a promising and important research field in translational medicine. Oncolytic viruses can induce immunogenic cell death by activating the body's immune system to recognize tumor cells. This work presents the results for optimizing the production of recombinant vesicular stomatitis viruses (rVSVs). To ensure the assembly of viral particles, we developed the HEK293TN-T7 cell line, which stably expresses DNA-dependent RNA polymerase 7 for viral genome transcription, and obtained helper plasmids encoding viral genes under the control of the CAG promoter. The oncolytic activity of the purified virus preparation was assessed in a murine model of B16F10Red melanoma cells expressing a red fluorescent protein. The presented method makes it possible to obtain purified viral preparations with a high titer and oncolytic activity. The amplification of viral particles in a HEK293 suspension culture allows for rapid scalability. Therefore, the developed approach can be used to obtain other recombinant VSV-based oncolytic viruses for tumor immunotherapy.

Transcriptional control of insulin-sensitive glucose carrier Glut4 expression in adipose tissue cells.

In search for new targets for obesity treatment, we have studied the effect of several transcription factors on the conversion of murine preadipocytes from the 3T3-L1 cell line into adipocytes. We have found that knockdown of Prep1 gene expression affects adipogenic differentiation and results in significant increase in the insulin-sensitive glucose carrier Glut4 gene expression.

[Enhancement of the medical care system for crews on space missions].

An overview of structural, operational and research aspects of the Russian system of medical support to health and performance of cosmonauts on the International space station (ISS) is presented. The backbone of the current tactics of cosmonauts' health maintenance is the original Russian medical care system developed for long-term piloted space fights. Over 12 years of its existence, the ISS has been operated by 33 main crews. The ISS program entrusted the established multilateral medical boards and panels with laying down the health standards as well as the generic and specific medical and engineering requirements mandatory to all international partners. Due to the program international nature, MedOps planning and implementation are coordinated within the network of working level groups with members designated by each IP. The article sums up the experiences and outlines future trends of the Russian medical care system for ISS cosmonauts. The authors pay tribute to academician Anatoli I. Grigoriev for his contribution to creation of the national system of medical safety in long-term piloted space missions, setting the ISS health and environmental standards and uniform principles of integrated crew health management, and gaining consensus on medical policy and operational issues equally during the ISS construction and utilization.

Insulin resistance and adipogenesis: role of transcription and secreted factors.

Insulin stimulates carbohydrate uptake by cells and induces their conversion into lipids as a more efficient form of energy storage. Insulin resistance is associated with a decrease in glucose uptake by muscle and adipose cells and also with a decrease in glycogen synthesis on retention of glucose synthesis by liver cells. Disorders in the insulin signaling cascade on development of insulin resistance can be caused by both changes in functioning of transcriptional factors and in the secretion profile of hormone-like substances. Diacylglycerols and ceramides responsible for activation of some kinases and phosphatases can directly trigger these changes in muscle and liver cells. In adipose tissue, insulin mainly stimulates adipogenesis (adipocyte differentiation) and lipogenesis (lipid accumulation in the cells). Thus, studies on the action mechanisms of factors influencing adipogenesis can be of help for understanding the molecular mechanisms of insulin resistance.

[Some approaches to the countermeasure system for a mars exploration mission].

In article discussed physiological and methodical principles of the organization of training process and his (its) computerization during Martian flight in conditions of autonomous activity of the crew, providing interaction with onboard medical means, self-maintained by crew of the their health, performance of preventive measures, diagnostic studies and, in case of necessity, carrying out of treatment. In super long autonomous flights essentially become complicated the control of ground experts over of crew members conditions, that testifies to necessity of a computerization of control process by a state of health of crew, including carrying out of preventive actions. The situation becomes complicated impossibility of reception and transfer aboard the necessary information in real time and emergency returning of crew to the Earth. In these conditions realization of problems of physical preventive maintenance should be solved by means of the onboard automated expert system, providing management by trainings of each crew members, directed on optimization of their psychophysical condition.

[Cluster analysis applicability to fitness evaluation of cosmonauts on long-term missions of the International space station].

Cluster analysis was applied to evaluate locomotion training (running and running intermingled with walking) of 13 cosmonauts on long-term ISS missions by the parameters of duration (min), distance (m) and intensity (km/h). Based on the results of analyses, the cosmonauts were distributed into three steady groups of 2, 5 and 6 persons. Distance and speed showed a statistical rise (p < 0.03) from group 1 to group 3. Duration of physical locomotion training was not statistically different in the groups (p = 0.125). Therefore, cluster analysis is an adequate method of evaluating fitness of cosmonauts on long-term missions.

[Quantitative evaluation of the effectiveness of physical contermeasures against motor disorders in long-duration space flight].

Regression analysis has been applied to determine the dependence of the motor system status (MSS) in long-duration space flight on the physical training programme (PTP), and flight duration (FD). Analyzed were graphical data presented in previous papers. Statistical analysis brought to light a high level of MSS-PTP relationship described as a 5-degree polynomial and a less explicit, yet statistically reliable MSS-FD relationship which is most adequately approximated by a 6-degree polynomial.

[Medical care for Russian cosmonauts' health on the ISS].

Established with the personal participation of O.G. Gazenko, the Russian system of medical care for cosmonauts' health has been largely preserved till this day. The system was fully functional on board the orbital complex MIR and, with appropriate modifications, has been adopted as a core of the medical care for Russian members of the ISS crews. In the period of 2000-2008, 22 cosmonauts were members of 17 ISS increments from 140 to 216 days in duration. The main functions of the medical care system were to control health, physical and mental performance, and to support implementation of space researches. The flow of readaptation to the normal gravity was, in most cases similar to what has been typical on return from the Russian orbital stations; some deviations are accounted for by application of the in-flight countermeasures. The paper familiarizes reader with some aspects of the theoretical work of academician O.G. Gazenko in the field of medical care in space flight. It outlines the principles of ISS medical management. The integrated medical support system combines medical equipment and items available on the Russian and US segments; the integrated medical group consists of flight surgeons, medical experts and biomedical engineers of the international partners and coordinates planning and implementation of medical operations. Also, challenges of health care on the phase of ISS utilization are defined.

[The countermeasure system for extended space flights].

The article summarizes the results of developing countermeasures by the team of winners of the USSR state prize under the leadership of academician O.G. Gazenko, and ensuing investigations at the Institute for Biomedical Problems. The system of countermeasures against the debilitating developments in cosmonaut's organism first developed in Russia ensured successful completion of long-term SF (64 to 438 days) aboard the Salyut and Mir orbital stations. The system incorporates exercises on the treadmill and veloergometer, axial body loading in suit Pinguin, application of negative pressure on the lower part of the body vacuum suit Chibis and several others. The system proved high efficiency in preventing or smoothing over the negative effects of microgravity in the course and after long-term space flight.

[Problems of diagnostics and prophylaxis in a Martian flight]. / Voprosy diagnostiki i profilaktiki v marsiankom polete.

In future interplanetary missions that will take up to two years the role of monitoring of the main human body systems and prevention of the effects of microgravity for the benefit of crew health and performance will rise eminently. In contrast to the orbital flights, diagnostics and prophylaxis will be a responsibility of the doctor within the crew who will consult solely own expertise and judgment. Hence, criteria and requirements to health monitoring and medicare systems and items are to be defined. The paper substantiates a concept and principles of diagnostics and prophylaxis, ideas concerning the scope of associated measures on the way to Mars, during the visit to the planet and on return to Earth; potentiality of a short-arch centrifuge as a means of prophylaxis is also discussed.

[Several clinical aspects of a piloted Martian flight]. / Nekotorye klinicheskie aspekty pilotiruemogo marsianskogo poleta.

The authors consider factors and risk sources in long and superlong (interplanetary) missions, conceivable effects of extended exposures in microgravity, and probability of diseases in the crew. They also lay down the basic principles of medical care, including surgery, to be applied in the interplanetary mission, and propose a nomenclature of key means and equipment for the therapeutic and emergency medical care, and a configuration of a medical compartment.

[An expert system for controlling the physical training program of crews on long-term space missions]. / Ekspertnaia sistema upravleniia fizicheskimi trenirovkami ékipazha v dlitel'nom kosmicheskom polete.

The concept of in-flight expert system for controlling (ESC) the physical training program during extended, including Martian, space missions has been developed based on the literature dedicated to the microgravity countermeasures and a retrospective analysis of effectiveness of the known ESC methods. This concept and the principle of crew autonomy were used as prime assumptions for defining the structure of ESC-based training in long-duration and planetary missions.

[Regulation of the human cardiovascular system under microgravitation]. / Reguliatsiia serdechno-sosudistoi sistemy cheloveka v usloviiakh mikrogravitatsii.

The paper presents the results of studies of the cardiovascular system and analyzes the mechanisms of regulation and adaptation of this system during long-term space flights. Special attention is given to the analysis of the homeostatic mechanisms for maintaining a number of integrative parameters of the cardiovascular system at the pre- and near-flight level and to the mechanisms of development of orthostatic detraining character after flights. How a new level of circulatory function in the absence of microgravitation is formed is shown.

[From 108 minutes to 438 days and further on... (on the 40th anniversary of Iu. A. Gagarin's flight)]. / Ot 108 minut do 438 sutok i dalee...(k 40-letiiu poleta Iu. A. Gagarina).

The progress of science and technology at the end of the XIXth and first half of the XXth century paved the way to start space exploration by humanity. The flight of Yu. A. Gagarin on April 12, 1961 was one of the history watersheds that had a great many of social implications. Piloted missions to space demonstrated the possibility for humans to adapt to the spaceflight factors which, nonetheless, can provoke various unfavorable reactions, particularly on return to Earth. Step-by-step extension of mission length paralleled enhancement of the methods of monitoring of crew health, life support systems, and development of modalities to maintain crew health and performance. The paper contains brief discussion of data acquired in short- and long-term missions including the 438-d mission of cosmonaut-physician V.V. Polyakov. The final section is devoted to the mainstream problems of future piloted space programs.

[Mechanisms reducing orthostatic stability of humans in long-term space flight]. / Mekhanizmy snizheniia ortostaticheskoi ustoichivosti v usloviiakh dlitel'nykh kosmicheskikh poletov.

Investigations conducted in and following space flights showed consistent decreases in cosmonauts' orthostatic stability due to the effects of long-term microgravity. Degree of this orthostatic disturbance depends on many factors prime of which is adequacy of countermeasures and least consequential is length of space flight, personal features, and others. Shifts in initial circulatory status and establishment of a new level of circulation and functioning of the other body systems in microgravity appear to play the central role in loss of orthostatic stability. The paper contains an in-depth analysis of processes occurring in human body during long-term stay in microgravity impacting orthostatic stability post flight.

[Some medical problems of piloted mission to Mars]. / Nekotorye meditsinskie problemy pilotiruemoi marsianskoi ekspeditsii.

Medical provisions to planetary missions take in a wide spectrum of medical, engineering and organizational issues the goal of which is to provide good living conditions for humans in space, and to ensure physical and psychic health and performance of crew members during research activities on the Martian surface and soon after return to Earth. The article carefully reviews the problems of defining conceptual approaches to medical support in a mission to Mars. These approaches are grounded on the analysis of medical experience of prolonged orbital missions and prognosis of physiological shifts due to extension of mission up to 2 to 3 years including the period of stay on the Martian surface. Special attention is given to the key medical aspects of the mission, i.e. medical monitoring, countermeasures (including utilization of a short-arm centrifuge) against the adverse effects of microgravity on crew health and keeping-up useful skills and knowledge.

[Use of academician P.K.Anokhin's creative heritage for analysis of mechanisms responsible for homeostasis formation under microgravity]. / Ispol'zovanie tvorcheskogo naslediia akademika P.K.Anokhina dlia analiza mekhanizmov formirovaniia gomeostaza v usloviiakh mikrogravitatsii.

The paper analyzes the changes occurring in the functions of the body's different systems under microgravitation in the context of current views on homeostasis and of the Anokhin's theory of functional systems. Proceeding from the Anokhin's views, the authors consider the specific features of a conceptual model for formation of the functional system of homeostatic regulation under microgravitation. These concepts were used to examine the body's homeostatic responses under the conditions of microgravitation which are associated with changes in the sensory input, with the movement of fluids in the cranial direction and with the removal of weight burden on the locomotor apparatus. Gravity withdrawal was demonstrated to lead to the development of homeostatic changes which are followed by consecutive trigger of prompt and long-term adaptation responses. This yields a new functional system of homeostatic regulation and a new rather relatively permanent functioning of the body's basic systems.

Manned interplanetary missions: prospective medical problems.

The present review aimed to suggest approaches to prospective medical problems related to the health maintenance of space crews during future manned interplanetary, particularly Martian, missions up to 2-3 years with a possible stay on a planet with gravity different from that on Earth. The approaches are based on knowledge so far obtained from our analysis of the medical support of long-term orbital flights up to one year, as well as on the consideration of specific conditions of interplanetary missions. These specific conditions include not only long-term exposure to microgravity, but also a prolonged stay of unpredictable duration (2-3 years) on board a spacecraft or on a planet without direct contact with Earth, and living in a team with a risk of psychological incompatibility and the impossibility of an urgent return to Earth. These conditions necessitate a highly trained medical person in the crew, diagnostic tools and equipment, psychophysiological support, countermeasures, as well as the means for urgent, including surgical, treatment on board a spacecraft or on a planet. In this review, the discussion was focused on the following predictable medical problems during an interplanetary mission; 1) unfavorable effects of prolonged exposure to microgravity, 2) specific problems related to Martian missions, 3) medical monitoring, 4) countermeasures, 5) psychophysiological support and 6) the medical care system.

[Mechanisms of homeostasis establishment during prolonged microgravity]. / Mekhanizmy formirovaniia gomeostaza pri dlitel'nom prebyvanii v usloviiakh mikrogravitatsii.

Investigations performed during prolonged microgravity were aimed at determination of phenomenology of the microgravity effects, and collection of data that would permit analysis of the micro-g specific homeostasis. As was stated, long-term microgravity alters the level of functioning of the main body systems, and a number of parameters of human metabolism and internal medium. It also instigates restructuring of some tissues and organs (primarily the musculoskeletal apparatus), brings along another level of energy and plastic (protein) metabolism, enhances the catabolic processes, and modifies the neuroendocrine regulation. In microgravity, the balance of functional loads on various body systems changes affecting rearrangement of the homeostasis regulation. The paper details the functional and morphological changes arising during long stay in microgravity and mechanisms culminating in a novel homeostasis in microgravity.