Analysis of PD-1 expression in the monocyte subsets from non-septic and septic preterm neonates

Programmed death-1 (PD-1) receptor system represents a part of recently reported immunoregulatory pathway. PD-1 is an immune checkpoint molecule, which plays an important role in downregulating the immune system proinflammatory activity. Until recently, PD-1 expression was not established on immune cells of the preterm infants. The study objectives were to confirm expression of the PD-1 receptors on the monocytes isolated from very low birth weight newborns (VLBW), and to analyze their expression during the first week of life and late-onset sepsis. Peripheral blood mononuclear cells were isolated from 76 VLBW patients without early-onset sepsis on their 5th day of life (DOL). PD-1 expression was determined on the monocyte subsets (classical, intermediate, non-classical) by flow cytometry. In case of late-onset sepsis (LOS), the same analysis was performed. Our results demonstrated that on the 5th DOL, PD-1 receptors were present in all the monocyte subsets. Children, whose mothers had received antenatal steroids, presented higher absolute numbers of non-classical monocytes with PD-1 expression. Infants born extremely preterm who later developed LOS, initially showed a lower percentage of PD-1 receptor-positive intermediate monocytes in comparison to neonates born very preterm. During LOS, we observed a rise in the percentage of classical monocytes with PD-1 expression. In case of septic shock or fatal outcome, there was a higher percentage and absolute count of intermediate monocytes with PD-1 expression in comparison to children without these complications. In conclusion, monocytes from VLBW children express PD-1 receptors. Antenatal steroid administration seems to induce PD-1 receptor expression in the non-classical monocytes. PD-1 might play a role in immunosuppressive phase of sepsis in the prematurely born children with septic shock and fatal outcome

Chronic mild stress alters the somatostatin receptors in the rat brain

RATIONALE: The involvement of somatostatin (SST) and its receptors in the pathophysiology of depression and stress has been evidenced by numerous studies. OBJECTIVES: The purpose of the present study was to find whether chronic mild stress (CMS), an animal model of depression, affects the SST receptors in the rat brain and pituitary, as well as the level of SST in plasma. METHODS: In CMS model, rats were subjected to 2 weeks of stress and behaviorally characterized using the sucrose consumption test into differently reacting groups based on their response to stress, i.e., stress-reactive (anhedonic), stress-non-reactive (resilient), and invert-reactive rats (characterized by excessive sucrose intake). We measured specific binding of [(125)I]Tyr(3)-Octreotide, expression of mRNA encoding sst2R receptors in the rat brains, expression of SST and its receptors in rat pituitary, and the level of SST in the plasma. RESULTS: The obtained results show decreases in binding of [(125)I]Tyr(3)-Octreotide in most of rat brain regions upon CMS and no significant differences between three stressed groups of animals, except for significant up-regulation of sst2 receptor in medial habenula (MHb) in the stress-reactive group. In the same group of animals, significant increase in plasma SST level was observed. CONCLUSIONS: There are two particularly sensitive sites distinguishing the response to stress in CMS model. In the brain, it is MHb, while on the periphery this predictor is SST level in plasma. These changes may broaden an understanding of the mechanisms involved in the stress response and point to the intriguing role of MHb

Emergent Role of IFITM1/3 towards Splicing Factor (SRSF1) and Antigen-Presenting Molecule (HLA-B) in Cervical Cancer

The IFITM restriction factors play a role in cancer cell progression through undefined mechanisms. We investigate new protein–protein interactions for IFITM1/3 in the context of cancer that would shed some light on how IFITM1/3 attenuate the expression of targeted proteins such as HLA-B. SBP-tagged IFITM1 protein was used to identify an association of IFITM1 protein with the SRSF1 splicing factor and transporter of mRNA to the ribosome. Using in situ proximity ligation assays, we confirmed a predominant cytosolic protein–protein association for SRSF1 and IFITM1/3. Accordingly, IFITM1/3 interacted with HLA-B mRNA in response to IFNγ stimulation using RNA–protein proximity ligation assays. In addition, RT-qPCR assays in IFITM1/IFITM3 null cells and wt-SiHa cells indicated that HLA-B gene expression at the mRNA level does not account for lowered HLA-B protein synthesis in response to IFNγ. Complementary, shotgun RNA sequencing did not show major transcript differences between IFITM1/IFITM3 null cells and wt-SiHa cells. Furthermore, ribosome profiling using sucrose gradient sedimentation identified a reduction in 80S ribosomal fraction an IFITM1/IFITM3 null cells compared to wild type. It was partially reverted by IFITM1/3 complementation. Our data link IFITM1/3 proteins to HLA-B mRNA and SRSF1 and, all together, our results begin to elucidate how IFITM1/3 catalyze the synthesis of target proteins. IFITMs are widely studied for their role in inhibiting viruses, and multiple studies have associated IFITMs with cancer progression. Our study has identified new proteins associated with IFITMs which support their role in mediating protein expression; a pivotal function that is highly relevant for viral infection and cancer progression. Our results suggest that IFITM1/3 affect the expression of targeted proteins; among them, we identified HLA-B. Changes in HLA-B expression could impact the presentation and recognition of oncogenic antigens on the cell surface by cytotoxic T cells and, ultimately, limit tumor cell eradication. In addition, the role of IFITMs in mediating protein abundance is relevant, as it has the potential for regulating the expression of viral and oncogenic proteins

Cholesterol Depletion in Adipocytes Causes Caveolae Collapse Concomitant with Proteosomal Degradation of Cavin-2 in a Switch-Like Fashion

Caveolae, little caves of cell surfaces, are enriched in cholesterol, a certain level of which is required for their structural integrity. Here we show in adipocytes that cavin-2, a peripheral membrane protein and one of 3 cavin isoforms present in caveolae from non-muscle tissue, is degraded upon cholesterol depletion in a rapid fashion resulting in collapse of caveolae. We exposed 3T3-L1 adipocytes to the cholesterol depleting agent methyl-β-cyclodextrin, which results in a sudden and extensive degradation of cavin-2 by the proteasome and a concomitant movement of cavin-1 from the plasma membrane to the cytosol along with loss of caveolae. The recovery of cavin-2 at the plasma membrane is cholesterol-dependent and is required for the return of cavin-1 from the cytosol to the cell surface and caveolae restoration. Expression of shRNA directed against cavin-2 also results in a cytosolic distribution of cavin-1 and loss of caveolae. Taken together, these data demonstrate that cavin-2 functions as a cholesterol responsive component of caveolae that is required for cavin-1 localization to the plasma membrane, and caveolae structural integrity

European External Action Service in view of Common Foreign and Security Policy

W swojej pracy staram się odpowiedzieć na pytanie, dlaczego zdecydowano się utworzyć Europejską Służbę Działań Zewnętrznych, w jaki sposób przebiegało jej tworzenie oraz jakie skutki się z tym wiążą dla Wspólnej Polityki Zagranicznej i Bezpieczeństwa.In my thesis I am trying to answer the question, why creation of European External Action Service is crucial for Common Foreign and Security Policy and how was the proccess of its creation

Norwegian foreign policy after 1990

The first chapter of this thesis addresses the issue of the Atlantic dimension in Norway's foreign policy. It contains an analysis of the way Norwegian economic interests associated with the exploitation of natural maritime resources are protected. The influence of geopolitical changes in the High North on Norway's safety and economic situation is studied afterwards, especially in view of the country's relations with its largest neighbor, Russia. Norway's NATO membership is subsequently presented as the main pillar of its security policy. The second chapter is dedicated to European integration in the Norwegian political economic and social discourse. It encompasses an attempt to answer the question why Norway hasn't joined the European Union and what once was and currently is the attitude towards the issue among the political elite as well as society. It also describes how strong the relationship between the European Union and Norway is, despite it remaining outside the Union's formal structures. The discussion in the third chapter revolves around Norway's foreign policy associated with its geographically, historically and culturally similar Scandinavian neighbors. The most important areas of the highly developed Scandinavian cooperation are presented along with the functioning of the Nordic Council as the best example of the integration and bilateral relations in that particular region. Finally, the fourth chapter is an analysis of Norway's policy of engagement and the causes of its implementation. It explains why Norway is regarded as a „moral superpower” in today's world and studies Norwegian efforts in building peace in the world, fighting poverty and promoting human rights.W rozdziale pierwszym niniejszej pracy poruszam kwestię wymiaru atlantyckiego w polityce zagranicznej Norwegii. Analizuję w jaki sposób chronione są norweskie interesy gospodarcze związane z eksploatacją bogactw naturalnych mórz. W dalszej kolejności zastanawiam się nad tym, jak zmiany geopolityczne na Dalekiej Północy wpływają na norweskie bezpieczeństwo i sytuację gospodarczą- analizuję stosunki tego kraju z jego największym sąsiadem- Rosją. Następnie przedstawiam norweskie uczestnictwo w NATO jako główny filar jej polityki bezpieczeństwa.Drugi rozdział poświęciłam integracji europejskiej w norweskim dyskursie politycznym, ekonomicznym i społecznym. Staram się odpowiedzieć na pytanie, dlaczego Norwegia nie przystąpiła do Unii Europejskiej oraz jaki był kiedyś i jaki jest obecnie stosunek do tego tematu wśród elit politycznych i wśród społeczeństwa. Przedstawiam także silne relacje łączące Norwegię z Unią Europejską, pomimo formalnego pozostawania poza jej strukturami. W trzecim rozdziale rozważam politykę zagraniczną Norwegii związaną z jej najbliższymi geograficznie, historycznie, kulturowo i językowo sąsiadami- państwami skandynawskimi. Przedstawiam najważniejsze obszary bardzo rozwiniętej współpracy państw skandynawskich oraz funkcjonowanie Rady Nordyckiej jako najlepszego przykładu integracji w regionie i wzajemnych relacji. W ostatnim, czwartym rozdziale, analizuję politykę zaangażowania Norwegii, zastanawiam się nad jej przyczynami oraz opisuję jak doszło do tego, że kraj ten uważany jest w dzisiejszym świecie za „moralne mocarstwo”. Analizuję norweskie działania na rzecz budowania pokoju na świecie, walki z ubóstwem, promowania praw człowieka

Implementation of the Results of Experimental Studies with the Use of the Sclerometric Method of Plane Elements in Wooden Buildings

Wood is one of the basic building materials. It is a completely biodegradable raw industrial commodity, the resources of which, with proper forest management, are virtually inexhaustible. Additionally, its acquisition and processing does not require large inputs of fossil fuels. At the same time, forest areas which we obtain wood from neutralize the negative effects of producing and acquiring other raw materials, as one hectare of pine forest (the most popular in Poland) can absorb approx. 20–30 tons of CO2. Wood is characterised by low thermal and electrical conductivity, having simultaneously high sound insulation, which perfectly meets the requirements of the present market and its regulations. This study aimed at verifying the technical parameters of wood, i.e., its bending strength, with the use of an innovative method of the correlation between the bending strength measured along and across wood fibres. The procedure was envisaged as effective for testing the strength of beams in historic buildings, in which—due to their valuable structure—only a limited number of sample holes can be made. The aim of this experiment was to create tables and diagrams, from which, based on the correlation between the side and the head of the beam, using in situ tests and the sclerometric method, it will be possible to derive the bending strength of existing wooden beams. In the study of spruce and pine wood, a correlation between the recess from the side and the recess from the head was found, ranging from 0.64 to 0.76, with an average of 0.72 for spruce elements, and 0.66–0.84, with an average of 0.70 for pine elements. This means that when testing an element fixed in a building, measuring the parameters from the head of the beam with a Schmidt hammer (often such elements are more easily accessible, i.e., on the building facade), the obtained values should be multiplied by 0.72 for spruce elements and by 0.70 for pine elements to obtain the strength of the beam. The authors of this article indicate that the confirmation of this observation requires conducting further research on various types of wood. It should also be noted that the material collected from one batch of sawn timber had a different structure, which was proved by analysing it using SEM imaging. Modeling wood numerically is, to some extent, a simplified issue that assumes wood to be an orthotropic, homogeneous (homogeneous) material. In fact, wood is an anisotropic, very heterogeneous material. The analysis of wood (on the technical scale, construction wood) as an anisotropic material is practically impossible. Adopting wood as an isotropic material is too simplistic. Therefore, the most appropriate methods of strength testing are destructive methods, as all non-destructive methods should not be used without verifying the results with other methods. The results obtained by non-destructive testing pose great difficulties in their interpretation. Obtaining reliable results of experiments entails collecting a large number of research samples. The method described in this paper will allow for obtaining the necessary data for effective expertise assessment regarding the safety level of structural elements in historic wooden load-bearing structures, which is crucial for making conservation decisions