Development of bispecific antibodies for selective stimulation of the CD95 death receptor on malignant and normal activated B-cells

Treatment of oncological and autoimmune diseases still remains a considerable challenge to modern medicine. Current therapy options such as chemotherapy and radiotherapy in oncology and glucocorticoids or other conventional immunosuppressive drugs for autoimmune diseases are often of limited efficacy. They suffer from a lack of specificity, with respect to the eradication of tumour and immune cells, respectively. The use of monoclonal antibodies in oncology allowed in some instances specific targeting of tumour cells and thus, improved this situation in the last decades. Some antibodies, such as anti-CD20 antibody Rituximab, targeting normal and malignant B-cells, are meanwhile firmly established in oncological treatment regimes. Interestingly, over the last years the anti B-cell activity of Rituximab was used in addition in numerous studies for the treatment of autoimmune diseases. Many strategies have been developed to further improve therapeutic activity of monoclonal antibodies. One of them is the development of bispecific antibodies recognising target cell antigens and effector cells or effector molecules at the same time. The “bispecific concept” used in this work is the target cell restricted stimulation of the death receptor CD95 (APO-1/Fas) with bispecific antibodies as described by Jung et al. in 2000. Whereas initial experiments were performed using chemically hybridised bispecific antibodies with CD20×CD95 specificity, the goal of this thesis was the generation of recombinant antibodies within a suitable format. To this end, a recombinant CD20×CD95 molecule in the so called FabSc format, designated BS9520 was developed and characterised in various in vitro and in vivo assays. It was found that the capability of this antibody to suppress the growth of malignant B-cells in vitro and in vivo and to specifically deplete normal, activated B-cells from PBMC cultures was superior to that achieved with monospecific clinically established anti-CD20 antibodies including a newly developed third generation Fc-optimized CD20 antibody. Moreover, the bispecific antibody was the only reagent capable of significantly suppressing IgG production by activated B-cells in vitro. These findings imply that the bispecific CD95×CD20 antibody might become an attractive reagent for the treatment of B-cell malignancies as well as B-cell mediated autoimmune diseases. In the second part of this work a completely new approach for the stimulation of the CD95 death receptor on antigen specific B-cells was evaluated. For proof of principle, the CD20 part in the BS9520 antibody was replaced by antigen fragments (tetanus or diphtheria toxoid derivatives). We named these fusion proteins BS95TT and BS95DT, respectively. Such protein should bind to B-cell receptor via their toxoid part and with the CD95 antibody part to CD95 receptor on activated TTspecific B-cells. Thereby, apoptosis is induced only in an antigen-specific fashion. This approach could be used for the treatment of autoimmune diseases if the antigen inducing auto-antibody production is known (such as Myasthenia gravis, multiple sclerosis and others)

Philosophy of Education as a Social Development Factor: World Trends and Prospects for Russia

The author analyzes the correlation between the development of society and philosophy of education. The goal of this article is to demonstrate that social development is more clearly shaped when the process of education is given an appropriate philosophy, which means the presence of a sense-making and goal-setting strategy (paradigm, “religion”) of the evolution of society. The example of a number of the world's leading countries confirms the value of such correlation. Even when in a crisis, these countries adapt to the modern world's dynamics (globalization, informatization, etc.) with higher speed if they possess an adequate education philosophy. These leading countries’ philosophy of education is underlain by the thesis: the quality of education defines the quality of life. This outcome can be achieved by drawing on such values as freedom, creativity, partnership, and trust. World university rankings show that the most successful universities aim at achieving this outcome and rely on these basic values regardless of all the modern transformations. The author comes to the conclusion that Russia, represented by its leaders and its government, should, given its desire to join the ranks of the world's leading countries, should instate substantial indicators of quality of life for its citizens, as well as strive to generate an education philosophy of similar nature and actively introduce it in the process of education in the first place

Respiratory Component of the Orienting Reflex: A Novel Sensitive Index of Sensory-Induced Arousal in Rats

In humans, the integrated response to a novel stimulus (orienting reflex, OR) includes behavioral (head turning etc.) and well-characterized physiological components (changes in heart rate, respiration, skin conductance, and EEG patterns). In rodents, the physiological components of the OR include changes in heart rate and cutaneous vasoconstrictor tone, but respiratory changes have so far not been systematically documented. In the present study conducted in adult male Wistar rats, the OR was elicited by 60-dB acoustic tones while animals were in a whole-body plethysmograph for respiratory recordings. In addition to respiration, in different groups of animals we concurrently recorded either EEG, or heart rate (both by biotelemetry), or tail blood flow (using ultrasound Doppler). Acoustic stimuli provoked vigorous tachypneic responses with respiratory rate rising from 80–100 to 450–650 cpm, and with small and variable changes in tidal volume. This respiratory arousal response was often, but not always, accompanied by EEG desynchronization and by variable tail vasoconstriction, and by small and inconsistent changes in the heart rate. We conclude that tachypneic responses are a new highly sensitive index of sensory-induced arousal

The role of GαO-mediated signaling in the rostral ventrolateral medulla oblongata in cardiovascular reflexes and control of cardiac ventricular excitability.

The heart is controlled by the sympathetic and parasympathetic limbs of the autonomic nervous system with inhibitory signaling mechanisms recruited in both limbs. The aim of this study was to determine the role of inhibitory heterotrimeric G proteins in the central nervous mechanisms underlying autonomic control of the heart and its potential role in arrhythmogenesis. Mice with conditional deletion of the inhibitory heterotrimeric G protein GαO in the presympathetic area of the rostral ventral lateral medulla (RVLM) were generated to determine the role of GαO-mediated signalling in autonomic control and electrophysiological properties of the heart. GαO deletion within the RVLM was not associated with changes in heart rate (HR) or the arterial blood pressure at rest (home cage, normal behavior). However, exposure to stressful conditions (novel environment, hypoxia, or hypercapnia) in these mice was associated with abnormal HR responses and an increased baroreflex gain when assessed under urethane anesthesia. This was associated with shortening of the ventricular effective refractory period. This phenotype was reversed by systemic beta-adrenoceptor blockade, suggesting that GαO depletion in the RVLM increases central sympathetic drive. The data obtained support the hypothesis that GαO-mediated signaling within the presympathetic circuits of the RVLM contributes to the autonomic control of the heart. GαO deficiency in the RVLM has a significant impact on cardiovascular responses to stress, cardiovascular reflexes and electrical properties of the heart.This research was supported by the Medical Research Council (MRC Clinical Research Training Fellowship to RA), British Heart Foundation (Ref: RG/14/4/30736), Wellcome Trust (Wellcome Trust Senior Research Fellowship to AVG; Ref: 095064), and by the Intramural Research Program of the National Institutes of Health, National Institute of Environmental Health Sciences (Project Z01- ES-101643 to LB). This work was facilitated by the National Institute for Health Research Barts Cardiovascular Biomedical Research Unit

COMMUNICATIVE COMPETENCE FORMATION OF BACHELORS MAJOR IN STATE AND MUNICIPAL MANAGEMENT

The article is devoted to a vital problem of modern Pedagogy that is to improve bachelor training in technical universities in conditions of new standards of higher education. The authors study the communicative competence that is one of the most important components of professional training of public service employees, and point to the need of adjusting the system of bachelor training, as well as creating a model of communicative competence

History of Russian Entrepreneurship in Signs of Commemoration: Formulating the Issue

The history of domestic business has been in focus of researches in recent decades. The problem of preserving the historical memory of Russian entrepreneurs in the socio-cultural environment of cities and villages of our country is becoming more and more important for both scientists and politicians. The authors introduce the idea of commemorative complex which is treated as a set of memory places associated with the activity of a certain representative of the commercial and industrial capital. This set is labeled one way or another. In spite of the fact that a considerable number of monuments, museums, sculptures is coming into being, memorial plaques keep a leading role among different ways of commemoration. Memorial plaques connected with the history of Russian entrepreneurship the authors divide into four groups: personal memorial plaques, memorial plaques dedicated to the business of charity, memorial plaques dedicated to the business functions, summarizing memorial plaques. The distinguished features of these types are analyzed. Memorial plaques help to recall the forgotten names of historical persons who did their best for the development and prosperity of their city and region. These signs of memory, as a rule, are an interesting source for studying the daily life of a certain city at a certain stage of its history. The article also presents the practical tasks arising from the study of memorial signs, dedicated to the prerevolutionary entrepreneurs and philanthropists. The role of modern business community in the process of the recovery of Russian entrepreneurs’ names into the political and social discourse is analyzed

Laser fusion of aluminum-alumina composites obtained in the vortex layer apparatus

The current study is devoted to laser fusion of aluminum-matrix composites with 3 wt. % of alumina obtained in the vortex layer apparatus. The mixing parameters and the distribution of alumina in the composite structure were studied. Three parameters of laser fusion for plate synthesis were used. The finest results were achieved using a volumetric energy density of 24 J/mm3 with 370 W laser powe