MANF as a new regulator of the unfolded protein response and maintenance of pancreatic β-cells in mice

Neurotrophic factors are small secretory proteins with essential roles in neuronal and non-neuronal tissues. Mesencephalic astrocyte-derived neurotrophic factor (MANF) and cerebral dopamine neurotrophic factor (CDNF) form a distinct family of unconventional neurotrophic factors. MANF and CDNF are endoplasmic reticulum (ER) located, but also secreted proteins. Initially, MANF was discovered as a trophic factor for dopamine neurons in vitro. Further studies revealed its protective and restorative properties in different animal disease models such as Parkinson´s disease, spinocerebellar ataxia, brain- and heart-ischemia. MANF is also identified as a protein upregulated in unfolded protein response (UPR) and protecting against ER stress-induced cell death. CDNF was identified based on its homology to MANF and characterized for its ability to protect and restore dopamine neurons in rodent models of Parkinson´s disease. However, the physiological roles of MANF and CDNF in mammals have remained unclear. The main objective of this thesis was, therefore, to study the biological roles of MANF in vivo by characterizing the phenotypes of MANF conventional and conditional knockout mice as well as analyzing MANF and CDNF expression in mouse tissues. Comprehensive expression analysis of MANF mRNA and protein revealed that MANF is widely expressed in most mouse tissues. It is highly expressed in neurons regulating energy homeostasis within the hypothalamus and neurons of other appetite-regulating areas including the brainstem structures and mesolimbic/mesocortical dopamine system. Exceptionally high levels of MANF was observed on peripheral mouse tissues with metabolic function, especially in cells with secretory functions within the endocrine and exocrine glands, suggesting essential roles for MANF in cells with high protein synthesis and secretion. Highest levels of CDNF protein was observed in tissues with high energy production and oxidative function including skeletal muscle, heart, testis and brown adipose tissue. In order to study the roles of MANF in mammals, we deloped MANF conventional knockout mice (Manf-/-), that showed severe growth retardation, poor survival and a progressive postnatal reduction of beta-cell mass resulting in severe insulin-deficient hyperglycemia and diabetes mellitus caused by decreased beta-cell proliferation and increased beta-cell apoptosis. In our further studies, we verified that diabetic phenotype of the Manf-/- mice was caused by a lack of MANF in the insulin-producing beta-cells in the pancreas and not in other organs by generating pancreas- and beta-cell-specific conditional Manf-/- mice. We found that pancreatic islets of conventional and conditional MANF deficient mice displayed chronic activation of the UPR, preceding downregulation of beta-cell markers, indicating unresolved ER stress as one possible cause of beta-cell failure in these mice. Thus, this work shows that MANF is an essential regulator of beta-cell maintenance and UPR in mice. We discovered that MANF protein increased beta-cell proliferation in vitro in islets isolated from young and even old mice. MANF protein also rescued mouse beta-cells from thapsigargin-induced apoptosis and ER stress-induced glucotoxicity in culture. Importantly, we found that MANF overexpression in the mouse pancreases mediated by adeno-associated virus vector was able to regenerate beta-cells in vivo in a mild low-dose streptozotocin mouse model of diabetes. Hence, these results indicate that MANF is a vital mitogen and protective protein for mouse beta-cells and can thus serve as a potential new regenerative drug for the diabetes therapy. Furthermore, we identified the decreased number of growth hormone (GH) and prolactin (PRL) expressing cells in the anterior pituitary gland of Manf-/- knockout mice associated with increased expression of UPR markers and decreased expression of Gh and Prl genes. Thus, reduced GH production could be one of the reasons for the growth retardation in Manf-/- mice. Taken together, the results in this thesis provide new critical biological functions of MANF in mouse in vivo which can be used to exploit the roles of MANF in human beta-cells and diabetes as well as in endocrine somatotropic cells and growth failure in human.Hermokasvutekijät ovat pieniä, solun erittämiä proteiineja, joilla on tärkeä rooli sekä hermostossa että muissa kudoksissa. Keskiaivojen astrosyyttiperäinen hermokasvutekijä (MANF) ja aivoperäinen dopamiinihermokasvutekijä (CDNF) muodostavat viimeisimpänä löydetyn epätavanomaisen hermokasvutekijäperheen. Niitä eritetään pieniä määriä, ja ne lokalisoituvat pääsääntöisesti solulimakalvostoon (engl. ER). Sekä CDNF- että MANF-proteiineilla on osoitettu olevan tärkeä rooli dopamiinihermosolujen kasvutekijänä Parkinsonin taudin eläinmalleissa. Lisäksi MANF suojaa soluja ER-stressin aiheuttamalta solukuolemalta. Tämän väitöskirjan päätavoitteena oli ymmärtää MANF-proteiinin biologista toimintamekanismia kuvaamalla kahta hiirimallia, joista MANF-geeni oli poistettu joko kudosspesifisesti tai kokonaan, sekä analysoida MANF- ja CDNF-proteiinien ilmentymistä hiiren kudoksissa. Tulokset osoittivat, että MANF mRNA:ta ja proteiinia löytyy suurimmasta osasta kudoksia, eniten hypotalamuksessa, aivorungossa ja dopamiinisoluissa. Lisäksi MANF-proteiinitasot ovat korkeat avo- ja umpirauhasissa. CDNF-proteiini puolestaan ilmentyy eniten luustolihaksissa, sydämessä, kiveksissä ja ruskeassa rasvakudoksessa. MANF-poistogeenisillä hiirillä havaittiin kasvun hidastuminen ja haiman insuliinia tuottavien beeta-solujen kuolemasta johtuva tyypin 1 diabetes, minkä vuoksi hiirten elinikä on keskimäärin alle kahdeksan viikkoa. Tutkimukset hiirillä, joilta MANF oli poistettu haimasta kudosspesifisesti, osoittivat että beeta-solujen kuolema johtuu jatkuvasta korkeasta solulimakalvoston stressitasosta, ja vahvistivat MANF-proteiinin tärkeän roolin beeta-soluja suojaavana kasvutekijänä sekä ER-stressin säätelijänä. Lisäksi osoitimme, että MANF-proteiini suojaa hiiren beeta-soluja thapsigarginilla indusoitulta solukuolemalta ja ER-stressin aiheuttamalta hyperglykemialta, sekä edistää beeta-solujen jakautumista nuorista ja myös vanhoista hiiristä eristetyissä Langerhansin saarekkeissa in vitro. Yliekspressoimalla MANF-proteiinia pystyimme suojaamaan ja regeneroimaan beeta-soluja myös in vivo streptozototsiinilla indusoidussa hiiren diabetes-mallissa. Selvitimme myös, että Manf-/- hiirien aivolisäkkeessä on vähemmän kasvuhormonia (GH) ja prolaktiinia tuottavia soluja, matalampi kasvuhormonin ja prolaktiinin geenien ekspressiotaso ja UPR-merkkiaineiden kohonnut ilmentyminen. Nämä löydöt voisivat osittain selittää, miksi Manf-poistogeeniset hiiret kasvavat hitaammin. Väitöskirjan tuloksena tiedämme enemmän MANF-proteiinin tärkeistä biologisista toimintamekanismeista hiirissä in vivo. Tätä tietoa voidaan hyödyntää jatkossa kuvattaessa MANF-proteiinin roolia ihmisen beeta-soluissa ja diabeteksessa, mutta myös tutkittaessa kasvun ja hormonitoiminnan häiriöitä

Massification of the Higher Education as a Way to Individual Subjective Wellbeing

The massification of the university education is inevitable process nowadays. On the one hand it allows solving the problem of knowledge generation and dissemination. On the other hand it helps individuals to achieve the subjective wellbeing, which is based on acquisition of knowledge that is necessary for the professional and individual orientation. In this article authors made an attempt to set tendency between individual's participation in the educational process and the acquisition of wellbeing's state. Taking into account modern trend of continuous education, authors as well suppose the existence of permanent wellbeing state. This statement is partially proven by social surveys and research's data, which has been conducted in Russia, Brazil, South Africa and other countries. Attractiveness and accessibility of education for people from different social groups determine the future level of public education. The rapid technological development of society, accessibility and mass character of educational programs, unconscious strivings of the major part of society to wellbeing must inevitably lead to the growth of average citizen's accomplishment

Local Pedagogical History in Training Bachelors of Education

the main aim of the study is to investigate the issues of pedagogy and technology of local history studies applied in training Bachelors of Education at a pedagogical University. The authors describe results of experimental work carried out at Stary Oskol branch of Belgorod State National Research University and approbation of local pedagogical history technology. Regarding the methods used in the study, we consider the training system containing local history concepts is made up by the following components: the training process, educational activities and research, teacher’s internship, extracurricular and outreach activities. The outcomes of the study demonstrate that local pedagogical history contributes significantly to training Bachelors of Education

Subjective well-being as a result of the realization of projects of the elderly's involvement into the social life

The paper analyses the subjective well-being and Successful Aging concepts. Unbiased analysis of such conditions is rather problematic by virtue of such facts as time-specific emotional state, climate of residence, culture, current developments and etc. The study concludes that there is no correlation between subjective well-being and economic health or objective socioeconomic factors. The subjective well-being is influenced in most cases by subjective factors: one’s health status satisfaction, financial situation, the level of self-reliance or independence, family interaction and other human relations. But the main factor is the recognition of their mission by the elderly which places the faith and hope in their future modeling. Two categories of the elderly are sorted out for the study: the involved and noninvolved in social activity. The research concludes that any formal or informal activity of the elderly is one of the main factors of the successful aging. Pursuit of activities is uncharacteristic for noninvolved in social activity elderly people. The paper analyses the projects organized and implemented by not only the elderly people themselves but also state and social structures.. Conducted analysis indicates that the level of subjective well-being of the two categories of the elderly is increasing as the result of these projects realization

Nouvelles et anciennes inégalités à la lumière d’une crise complexe: le contexte de l’exclusion liée au logement et celui des réfugiés et des migrations New-old inequalities in light of the double crisis: the context of homelessness and refugee and migrat

The text will present the complex contexts of the social situation currently prevailing in Poland. Firstly, these are the migration effects of the war on February 24, 2022, when Russia invaded Ukraine. Secondly, it is a crisis related to war refugees (from Iran, Afghanistan, Syria, and various African countries) on the Polish-Belarusian border (since 2021). Both crises are covered in thoroughly different ways by the media, state, and public opinion. In these cases, we are dealing with a situation that does not create a homogenous image of migration and refugees, neither at the level of social assessments nor when it comes to the law or the state support system. To understand the situation, it is necessary to observe the development of non-institutional and grassroots forms of approach to social and humanitarian crises in Polish society, including those led by NGOs. However, the models of assistance used for new groups in crisis in Poland (war refugees) are largely based on the well-known rules of assistance (homelessness). The question is: to what extent are the two crises compatible so that it is possible to use one type of experience (help in homelessness) to organize support for people remaining in another crisis (war refugees)? This is important in the Polish case, although it applied there the system of public social assistance, the authorities use only methods of managing crises and social problems, and not solving them

Nouvelles et anciennes inégalités à la lumière d’une crise complexe: le contexte de l’exclusion liée au logement et celui des réfugiés et des migrations New-old inequalities in light of the double crisis: the context of homelessness and refugee and migrat

The text will present the complex contexts of the social situation currently prevailing in Poland. Firstly, these are the migration effects of the war on February 24, 2022, when Russia invaded Ukraine. Secondly, it is a crisis related to war refugees (from Iran, Afghanistan, Syria, and various African countries) on the Polish-Belarusian border (since 2021). Both crises are covered in thoroughly different ways by the media, state, and public opinion. In these cases, we are dealing with a situation that does not create a homogenous image of migration and refugees, neither at the level of social assessments nor when it comes to the law or the state support system. To understand the situation, it is necessary to observe the development of non-institutional and grassroots forms of approach to social and humanitarian crises in Polish society, including those led by NGOs. However, the models of assistance used for new groups in crisis in Poland (war refugees) are largely based on the well-known rules of assistance (homelessness). The question is: to what extent are the two crises compatible so that it is possible to use one type of experience (help in homelessness) to organize support for people remaining in another crisis (war refugees)? This is important in the Polish case, although it applied there the system of public social assistance, the authorities use only methods of managing crises and social problems, and not solving them

Innovative Approaches in the Labor Organization of the Auxiliary Production at Mining and Smelting Enterprises

Application of work standardizing in the optimization of the number of personnel to improve production efficiency is now an example of the innovative approach to the organization of labour of auxiliary workers. In this article we propose an algorithm for calculating the planned number of repair workers based on the application of the time and labour content standards.Применение нормирования труда в рамках оптимизации численности персонала для по- вышения эффективности производства является в настоящее время примером иннова- ционного подхода в организации труда вспомогательных рабочих. В статье предлагается алгоритм расчета плановой численности ремонтных рабочих, основанный на применении норм времени и нормативов трудоемкости

MANF is Required for the Postnatal Expansion and Maintenance of the Pancreatic β-Cell Mass in Mice

Global lack of mesencephalic astrocyte-derived neurotropic factor (MANF) leads to progressive postnatal loss of β-cells mass and insulin-dependent diabetes in mice. Similarly to Manf -/- mice, embryonic ablation of MANF specifically from the pancreas results in diabetes. In this study, we assessed the importance of MANF for the postnatal expansion of the pancreatic β-cell mass and for adult β-cell maintenance in mice. Detailed analysis of Pdx-1Cre +/- ::Manf fl/fl mice revealed mosaic MANF expression in postnatal pancreases and significant correlation between the number of MANF-positive β-cells and β-cell mass in individual mice. In vitro, recombinant MANF induced β-cell proliferation in islets from aged mice and protected from hyperglycemia-induced endoplasmic reticulum (ER) stress. Consequently, excision of MANF from β-cells of adult MIP-1Cre ERT ::Manf fl/fl mice resulted in reduced β-cell mass and diabetes caused largely by β-cell ER stress and apoptosis, possibly accompanied by β-cell de-differentiation and reduced rates of β-cell proliferation. Thus, MANF expression in adult mouse β-cells is needed for their maintenance in vivo. We also revealed a mechanistic link between ER stress, and inflammatory signaling pathways leading to β-cell death in the absence of MANF. Hence, MANF might be a potential target for regenerative therapy in diabetes.Peer reviewe

Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF) Is Highly Expressed in Mouse Tissues With Metabolic Function

Peer reviewe

Russian sport economy: challenges and contradictions

The article shows that development of sport tends to be a factor of country competitiveness. Nowadays, there is a need to find new forms and methods of promotion and popularization of physical culture and sport, to create a single sports information space and to provide sports area with more effective management decisions. There were revealed the main challenges in sports area such as the difference of sports facilities in various regions of Russia, the creation of low-budget sports facilities, the development of qualitative modern material and technical base in educational institutions and another issue is qualified personnel training. Methods of achievement of the goals of strategy 2020 are also illustrated in the paper, that is getting large and medium-sized business involved in construction and operation of sports facilities as social one, creation of a single sports information space in order to control the workload of all the sports venues despite the fact of department affiliation