Fusion protein strategies for cryo-EM study of G protein-coupled receptors

Single particle cryogenic-electron microscopy (cryo-EM) is used extensively to determine structures of activated G protein-coupled receptors (GPCRs) in complex with G proteins or arrestins. However, applying it to GPCRs without signaling proteins remains challenging because most receptors lack structural features in their soluble domains to facilitate image alignment. In GPCR crystallography, inserting a fusion protein between transmembrane helices 5 and 6 is a highly successful strategy for crystallization. Although the similar strategy has the potential to broadly facilitate cryo-EM structure determination of GPCRs alone without signaling protein, the critical determinants that make this approach successful are not yet clear. Here, we address this shortcoming by exploring different fusion protein designs, which led to structures of antagonist bound A2A adenosine receptor at 3.4&Aring resolution and unliganded Smoothened at 3.7&Aring resolution. The fusion strategies explored here are likely applicable to cryo-EM interrogation of other GPCRs and small integral membrane proteins.

Oscillatory electroencephalographic patterns of arithmetic problem solving in fourth graders

Numerous studies have identified neurophysiological correlates of performing arithmetic in adults. For example, oscillatory electroencephalographic (EEG) patterns associated with retrieval and procedural strategies are well established. Whereas fact retrieval has been linked to enhanced left-hemispheric theta ERS (event-related synchronization), procedural strategies are accompanied by increased bilateral alpha ERD (event-related desynchronization). It is currently not clear if these findings generalize to children. Our study is the first to investigate oscillatory EEG activity related to strategy use and arithmetic operations in children. We assessed ERD/ERS correlates of 31 children in fourth grade (aged between nine and ten years) during arithmetic problem solving. We presented multiplication and subtraction problems, which children solved with fact retrieval or a procedure. We analyzed these four problem categories (retrieved multiplications, retrieved subtractions, procedural multiplications, and procedural subtractions) in our study. In summary, we found similar strategy-related patterns to those reported in previous studies with adults. That is, retrieval problems elicited stronger left-hemispheric theta ERS and weaker alpha ERD as compared to procedural problems. Interestingly, we observed neurophysiological differences between multiplications and subtractions within retrieval problems. Although there were no response time or accuracy differences, retrieved multiplications were accompanied by larger theta ERS than retrieved subtractions. This finding could indicate that retrieval of multiplication and subtraction facts are distinct processes, and/or that multiplications are more frequently retrieved than subtractions in this age group.

Uptake of biosimilar trastuzumab in Denmark compared with other European countries: a comparative study and discussion of factors influencing implementation and uptake of biosimilars

Purpose The aim of this study was to describe the implementation and uptake of biosimilar trastuzumab in Denmark compared with other European countries. Methods European data for usage of trastuzumab was supplied by IQVIA™, using the MIDAS® dataset. A comparison was performed based on market share estimated in sales volume. A separate comparison was undertaken between countries with a full two-fold switch between different biosimilars. Data was collected spanning the time from first registered sales of biosimilar trastuzumab until the 1st quarter of 2020. Results Denmark had the fastest and most thorough uptake of biosimilar trastuzumab compared with other EU countries. After 3 months, the market share of biosimilar trastuzumab had increased to 90% while the second fastest country had a 50% market share after 3 months. Only two other countries had undergone a full second switch between biosimilars, Hungary and Norway. All of the three countries made near complete switches between biosimilars while only Denmark had reduced the use of biooriginator below 10%. Conclusion The implementation of biosimilar trastuzumab in Denmark was rapid and achieved high overall uptake compared with other EU countries. The switch from one biosimilar to another was also achieved quickly and thoroughly. We believe that the rapid dissemination of information and involvement of all stakeholders — administrators, pharmacies, prescribers, nurses, and patients — constitute the backbone of the Danish success. A similar strategy is recommend for biosimilar implementation in other countries.

Oscillatory Electroencephalographic Patterns of Arithmetic Problem Solving in Fourth Graders

Numerous studies have identified neurophysiological correlates of performing arithmetic in adults. For example, oscillatory electroencephalographic (EEG) patterns associated with retrieval and procedural strategies are well established. Whereas fact retrieval has been linked to enhanced left-hemispheric theta ERS (event-related synchronization), procedural strategies are accompanied by increased bilateral alpha ERD (event-related desynchronization). It is currently not clear if these findings generalize to children.Our study is the first to investigate oscillatory EEG activity related to strategy use and arithmetic operations in children. We assessed ERD/ERS correlates of 31 children in fourth grade (aged between nine and ten years) during arithmetic problem solving. We presented multiplication and subtraction problems, which children solved with fact retrieval or via a procedure. Based on both problem size and verbal strategy reports, we analyzed these problem types separately for each operation.We found similar strategy-related patterns to those reported in previous studies with adults. That is, retrieval problems elicited stronger left-hemispheric theta ERS and weaker alpha ERD as compared to procedural problems. Interestingly, we observed differences between multiplications and subtractions within retrieval problems. Although there were no response time and accuracy differences, retrieved multiplications were accompanied by larger theta ERS than retrieved subtractions. This finding could indicate that retrieval of multiplication and subtraction facts are distinct processes, and/or that multiplications are more frequently retrieved than subtractions in this age group.

Antiviral defense via nucleotide depletion in bacteria

DNA viruses and retroviruses need to consume large quantities of deoxynucleotides (dNTPs) when replicating within infected cells. The human antiviral factor SAMHD1 takes advantage of this vulnerability in the viral life cycle, and inhibits viral replication by degrading dNTPs into their constituent deoxynucleosides and inorganic phosphate. In this study we report that bacteria employ a similar strategy to defend against phage infection. We found a family of defensive dCTP deaminase proteins that, in response to phage infection, convert dCTP into deoxy-uracil nucleotides. A second family of phage resistance genes encode dGTPase enzymes, which degrade dGTP into phosphate-free deoxy-guanosine (dG) and are distant homologs of the human SAMHD1. Our results show that the defensive proteins completely eliminate the specific deoxynucleotide (either dCTP or dGTP) from the nucleotide pool during phage infection, thus starving the phage of an essential DNA building block and halting its replication. Both defensive genes are found in a diverse set of bacterial species and are specifically enriched in Vibrio genomes. Our study demonstrates that manipulation of the deoxynucleotide pool is a potent antiviral strategy shared by both prokaryotes and eukaryotes.

World or event? To the ontology of war

This paper is an attempt to rethink the ontology of war. Its main object is to determine the ontological status of war and the connection between strategies of armed conflict prevention and the way this status is understood. If to overcome metaphysics we need to reconsider its basis then maybe a similar strategy should be applied in order to overcome war. The source of war is understood here not only in temporal terms, but also as its essence – the ontological basis. The first part of the article invokes several conceptions from the twentieth century (that, of course, refer to much older texts), according to which, war was to be overcome by variously understood progress. In those conceptions, as well as in the critical approaches from the beginning of the 21st century, war is treated as an event, a state of affairs. In the next part of my paper, based on Margaret Mead's anthropological diagnosis, I propound a conception of war as an invention, and therefore – technology. The issue of war is further considered in the context of the twentieth century philosophy of technology. A thesis is put forward according to which understanding the influence of techne of war over episteme should be the essence of thinking of war prevention. In this study, war is treated as a technology that determines perception. For critical reflection upon war, I use twentieth century philosophical conceptions linking technology with cognitive processes, especially those formulated by Martin Heidegger and Marshall McLuhan. Woniak A. wiat czy wydarzenie? W stron ontologii wojny // Argument. 2020. Vol. 1/10. Pp. 133-150.

The Effect of a Similar Strategy in The Rebuilding of Valuable Buildings

Reconstruction of archaeological buildings with destroyed or collapsed parts is considered oneof the most important and accurate operations in the field of preserving those buildings, given the continuityof these operations in the continuity of those archaeological buildings with their architectural and technicaldetails, and even their continued performance in their urban environment in many cases. These operationsare concerned with missing parts or the replacement of damaged parts. Many studies emerged that dealtwith this important aspect of reconfiguring and completing the demolished parts in historical and heritagebuildings, whether at the level of the part or all according to specific laws, but no study of the formal andintellectual mechanisms used to rebuild and reconstruct these buildings and their potential for architecturalproductions. Therefore, the research problem that emerged from the lack of clarity of knowledge about themost important formal and intellectual mechanisms for rebuilding in architecture and its cognitive indicatorswithin the outcome emerged, and the research was divided into two parts, the first part represented by thegeneral theoretical presentation that included the reconstruction processes and related determinants andclarification of the mechanism of formalism (similar and its connections) Theory and intellectual mechanism(analogy and its theoretical correlations), previous studies and the second part which represented the appliedside to form the theoretical framework from the above and applied it to groups of architectural productionsand then presented and discussed and analyzed the results of that application up to the presentation ofCrowns and recommendations that were referring to the general vision of the basic concept of researchadopted, the mechanistic aspect of form rather than intellectual.

Exposure to One Antibiotic Leads to Acquisition of Resistance to Another Antibiotic via Quorum Sensing Mechanisms

The vancomycin-resistant Enterococci (VRE) have progressively become a severe medical problem. Although clinics have started to reduce vancomycin prescription, vancomycin resistance has not been contained. We found that the transfer of vancomycin resistance in Enterococcus faecalis increased more than 30-fold upon treatment by streptomycin. Notably, treatment with an antibiotic caused the bacteria to become resistant to another. The response was even stronger in the well-studied plasmid pCF10 and the number of transconjugants increased about 100,000-fold. We tested four different antibiotics, and all of them induced conjugal response. Through a mathematical model based on gene regulation, we found a plausible explanation. Via quorum sensing, the change of the cell density triggers the conjugation. Moreover, we searched for generality and found a similar strategy in Bacillus subtilis. The outcome of the present study suggests that even common antibiotics must not be overused.

Świat czy wydarzenie? W stronę ontologii wojny

The world or an event? Towards an ontology of war: This paper is an attempt to rethink the ontology of war. Its main object is to determine the ontological status of war and the connection between strategies of armed conflict prevention and the way this status is understood. If to overcome metaphysics we need to reconsider its basis then maybe a similar strategy should be applied in order to overcome war. The source of war is understood here not only in temporal terms, but also as its essence — the ontological basis. The first part of the article invokes several conceptions from the twentieth century (that, of course, refer to much older texts), according to which, war was to be overcome by variously understood progress. In those conceptions, as well as in the critical approaches from the beginning of the 21st century, war is treated as an event, a state of affairs. In the next part of my paper, based on Margaret Mead’s anthropological diagnosis, I propound a conception of war as an invention, and therefore — technology. The issue of war is further considered in the context of the twentieth-century philosophy of technology. A thesis is put forward according to which understanding the influence of techne of war over episteme should be the essence of thinking of war prevention. In this study, war is treated as a technology that determines perception. For critical reflection upon war, I use twentieth-century philosophical conceptions linking technology with cognitive processes, especially those formulated by Martin Heidegger and Marshall McLuhan.

A Multimodal Discourse Analysis in Media

This study attempts a multimodal discourse analysis of a newspaper article during the Brexit campaign. The aim is to help the audience decode and evaluate photographs and texts from media by providing a few strategies as guidance. The approach adopted for this analytical research is inspired by Halliday and Matthiessen’s An Introduction to Functional Grammar (2004). Consequently, the objects in a photograph and words in a text function similarly conveying information to the audience, and additionally, involve a similar strategy of analysis. To sum up, these strategies are intended to improve the audience’s comprehension of decoding article meaning and journalist’s intention.