The glycine receptor alpha 3 subunit mRNA expression shows sex-dependent differences in the adult mouse brain

BackgroundThe glycinergic system plays an important inhibitory role in the mouse central nervous system, where glycine controls the excitability of spinal itch- and pain-mediating neurons. Impairments of the glycine receptors can cause motor and sensory deficits. Glycine exerts inhibition through interaction with ligand-gated ion channels composed of alpha and beta subunits. We have investigated the mRNA expression of the glycine receptor alpha 3 (Glra3) subunit in the nervous system as well as in several peripheral organs of female and male mice.ResultsSingle-cell RNA sequencing (scRNA-seq) data analysis on the Zeisel et al. (2018) dataset indicated widespread but low expression of Glra3 in vesicular glutamate transporter 2 (Vglut2, Slc17a6) positive and vesicular inhibitory amino acid transporter (Viaat, Slc32a1)positive neurons of the mouse central nervous system. Highest occurrence of Glra3 expression was identified in the cortex, amygdala, and striatal regions, as well as in the hypothalamus, brainstem and spinal cord. Bulk quantitative real-time-PCR (qRT-PCR) analysis demonstrated Glra3 expression in cortex, amygdala, striatum, hypothalamus, thalamus, pituitary gland, hippocampus, cerebellum, brainstem, and spinal cord. Additionally, male mice expressed higher levels of Glra3 in all investigated brain areas compared with female mice. Lastly, RNAscope spatially validated Glra3 expression in the areas indicated by the single-cell and bulk analyses. Moreover, RNAscope analysis confirmed co-localization of Glra3 with Slc17a6 or Slc32a1 in the central nervous system areas suggested from the single-cell data.ConclusionsGlra3 expression is low but widespread in the mouse central nervous system. Clear sex-dependent differences have been identified, indicating higher levels of Glra3 in several telencephalic and diencephalic areas, as well as in cerebellum and brainstem, in male mice compared with female mice

Robust T cell immunity in convalescent individuals with asymptomatic or mild COVID-19

SARS-CoV-2-specific memory T cells will likely prove critical for long-term immune protection against COVID-19. Here, we systematically mapped the functional and phenotypic landscape of SARS-CoV-2-specific T cell responses in unexposed individuals, exposed family members, and individuals with acute or convalescent COVID-19. Acute-phase SARS-CoV-2-specific T cells displayed a highly activated cytotoxic phenotype that correlated with various clinical markers of disease severity, whereas convalescent-phase SARS-CoV-2-specific T cells were polyfunctional and displayed a stem-like memory phenotype. Importantly, SARS-CoV-2-specific T cells were detectable in antibody-seronegative exposed family members and convalescent individuals with a history of asymptomatic and mild COVID-19. Our collective dataset shows that SARS-CoV-2 elicits broadly directed and functionally replete memory T cell responses, suggesting that natural exposure or infection may prevent recurrent episodes of severe COVID-19

Interventioner: ett skydd för mänskliga rättigheter eller en kränkning av internationell lagstiftning?

I den vetenskapliga artikeln ”The Lady Doth Proteset too Much: Kosovo and the turn of Ethics in international law”, presenterar Martti Koskenniemi en teori, gällande hur den internationella juristen använder moral som ett argument för att legitimera humanitära interventioner. Faran i detta blir enligt Koskenniemi att stater som redan besitter makt och militära resurser kommer använda lagen som ett instrument för att bedriva sina politiska intressen. Detta blir möjligt vid situationer där grova kränkningar mot mänskliga rättigheter sker och ingen längre kan ifrågasätta det moraliska behovet av att agera mot dessa kränkningar. Denna uppsats kommer vara en teoriprövning med bakgrund mot NATOs intervention i Kosovo 1999. Teoriprövningen kommer ske genom en argumentationsanalys av teser som presenteras i Koskenniemis artikel. Dessa teser kommer sedan jämföras med argument som presenteras i officiella uttalanden från NATO för att se om det finns något som talar för eller mot tesen i dessa uttalanden. Intervention, moral, NATO, legitimera, kränkningar, politiska intressen

Legitimitet genom den språkliga diskursen : En diskursanalys av Global Times och Hong Kong Free Press gällande protesterna i Hongkong 2019

Denna kandidatuppsats är en diskursanalys av den Kina-ägda tidningen Global Times och Hongkong-ägda Hong Kong Free Press gällande artiklar relaterade till de pågående protesterna i Hongkong. Konflikten tycks ha delats upp i två läger ett som representerar ”fastlands Kina” och ett som representerar ”Hongkongborna”. Uppsatsen syfte är att analyser den diskurs genom vilket de båda parterna försöker legitimera sin bild av kon-flikten för omvärlden. Detta kommer göras genom att analysera diskursen som presente-ras i respektive media utifrån fyra nyckelbegrepp inom konflikten, dessa är: ”Human Rights”. ”Hong Kong Human Right and Democracy Act of 2019”, ”Protests” och ”Ba-sic Law”. För att kunna ge en så aktuell bild av konflikten med hänsyn till uppsatsen deadline, har artiklarna jag jobbar utifrån begränsats till novembermånad. Frågeställningen som undersökningen kommer att utgå ifrån är: Vilka sanningsnar-rativ presenteras kring protesterna i medierna Global Times och Hong Kong Free Press och vilka skillnader och gemensamma utgångpunkter finns i de båda narrati-ven? Diskursanalysen kommer kombineras med teorin ”hegemoni” detta för att visa hur Global Times och Hong Kong Free Press försöker skapa en hegemonisk intervention genom respektive diskurs. Resultatet av undersökningen var att de olika diskurser som Global Times och Hong Kong Free Press skapat två verklighetsuppfattningar som konstant kolliderar med varandra. Bristen på en delad uppfattning av betydelserna för nyckelbegrep-pen, presenterade i ovanstående stycke, har skapat olika sanningsnarrativ mellan de båda medierna

Implementering av affärskoder i medarbetarnas dagliga arbete

While the stakeholders’ awareness about how companies act sustainably has increased, the business code has gained greater importance. With a functioning business code in a company, it can help guide the behavior of its employees to maintain an ethical company culture. In the study, the companies’ implementation of the business code in the employees’ daily work based on ethical core values, a formal ethics program and ethical leadership has been examined. As previous research in the field deals with whether there is a business code or not, and whether it has an effect on the employees, it is therefore interesting to investigate in this study how the business code is actually implemented in daily operations and whether companies make full use of the business code.  In order to achieve the purpose of the study, the study has targeted larger companies with a clearly designed business code. The data collection consists of 10 interviews with respondents who are familiar with the business code. The theory consists of previous research in CSR and about which approach is required to implement the business code in daily work. Research in ethical core values, a formal ethics program and ethical leadership are also mentioned.  The result of this study shows that more emphasis is placed on the ethical core values and ethical leadership and that the business code is overshadowed. On the other hand, the business code is included indirectly in the employees’ daily with through everyday communication around the ethical core values and a value-driven leadership. The study also contributes to an understanding of how the three different areas complement each other and highlights the area’s complexity.

Implementering av affärskoder i medarbetarnas dagliga arbete

While the stakeholders’ awareness about how companies act sustainably has increased, the business code has gained greater importance. With a functioning business code in a company, it can help guide the behavior of its employees to maintain an ethical company culture. In the study, the companies’ implementation of the business code in the employees’ daily work based on ethical core values, a formal ethics program and ethical leadership has been examined. As previous research in the field deals with whether there is a business code or not, and whether it has an effect on the employees, it is therefore interesting to investigate in this study how the business code is actually implemented in daily operations and whether companies make full use of the business code.  In order to achieve the purpose of the study, the study has targeted larger companies with a clearly designed business code. The data collection consists of 10 interviews with respondents who are familiar with the business code. The theory consists of previous research in CSR and about which approach is required to implement the business code in daily work. Research in ethical core values, a formal ethics program and ethical leadership are also mentioned.  The result of this study shows that more emphasis is placed on the ethical core values and ethical leadership and that the business code is overshadowed. On the other hand, the business code is included indirectly in the employees’ daily with through everyday communication around the ethical core values and a value-driven leadership. The study also contributes to an understanding of how the three different areas complement each other and highlights the area’s complexity.

The glycine receptor alpha 3 subunit mRNA expression shows sex-dependent differences in the adult mouse brain

Abstract Background The glycinergic system plays an important inhibitory role in the mouse central nervous system, where glycine controls the excitability of spinal itch- and pain-mediating neurons. Impairments of the glycine receptors can cause motor and sensory deficits. Glycine exerts inhibition through interaction with ligand-gated ion channels composed of alpha and beta subunits. We have investigated the mRNA expression of the glycine receptor alpha 3 (Glra3) subunit in the nervous system as well as in several peripheral organs of female and male mice. Results Single-cell RNA sequencing (scRNA-seq) data analysis on the Zeisel et al. (2018) dataset indicated widespread but low expression of Glra3 in vesicular glutamate transporter 2 (Vglut2, Slc17a6) positive and vesicular inhibitory amino acid transporter (Viaat, Slc32a1)positive neurons of the mouse central nervous system. Highest occurrence of Glra3 expression was identified in the cortex, amygdala, and striatal regions, as well as in the hypothalamus, brainstem and spinal cord. Bulk quantitative real-time-PCR (qRT-PCR) analysis demonstrated Glra3 expression in cortex, amygdala, striatum, hypothalamus, thalamus, pituitary gland, hippocampus, cerebellum, brainstem, and spinal cord. Additionally, male mice expressed higher levels of Glra3 in all investigated brain areas compared with female mice. Lastly, RNAscope spatially validated Glra3 expression in the areas indicated by the single-cell and bulk analyses. Moreover, RNAscope analysis confirmed co-localization of Glra3 with Slc17a6 or Slc32a1 in the central nervous system areas suggested from the single-cell data. Conclusions Glra3 expression is low but widespread in the mouse central nervous system. Clear sex-dependent differences have been identified, indicating higher levels of Glra3 in several telencephalic and diencephalic areas, as well as in cerebellum and brainstem, in male mice compared with female mice

Serine Protease Inhibitors Restrict Host Susceptibility to SARS-CoV-2 Infections

Identification of host factors affecting individual SARS-CoV-2 susceptibility will provide a better understanding of the large variations in disease severity and will identify potential factors that can be used, or targeted, in antiviral drug development. With the use of an advanced lung cell model established from several human donors, we identified cellular protease inhibitors, serpins, as host factors that restrict SARS-CoV-2 infection. The coronavirus disease 2019, COVID-19, is a complex disease with a wide range of symptoms from asymptomatic infections to severe acute respiratory syndrome with lethal outcome. Individual factors such as age, sex, and comorbidities increase the risk for severe infections, but other aspects, such as genetic variations, are also likely to affect the susceptibility to SARS-CoV-2 infection and disease severity. Here, we used a human 3D lung cell model based on primary cells derived from multiple donors to identity host factors that regulate SARS-CoV-2 infection. With a transcriptomics-based approach, we found that less susceptible donors show a higher expression level of serine protease inhibitors SERPINA1, SERPINE1, and SERPINE2, identifying variation in cellular serpin levels as restricting host factors for SARS-CoV-2 infection. We pinpoint their antiviral mechanism of action to inhibition of the cellular serine protease, TMPRSS2, thereby preventing cleavage of the viral spike protein and TMPRSS2-mediated entry into the target cells. By means of single-cell RNA sequencing, we further locate the expression of the individual serpins to basal, ciliated, club, and goblet cells. Our results add to the importance of genetic variations as determinants for SARS-CoV-2 susceptibility and suggest that genetic deficiencies of cellular serpins might represent risk factors for severe COVID-19. Our study further highlights TMPRSS2 as a promising target for antiviral intervention and opens the door for the usage of locally administered serpins as a treatment against COVID-19. IMPORTANCE Identification of host factors affecting individual SARS-CoV-2 susceptibility will provide a better understanding of the large variations in disease severity and will identify potential factors that can be used, or targeted, in antiviral drug development. With the use of an advanced lung cell model established from several human donors, we identified cellular protease inhibitors, serpins, as host factors that restrict SARS-CoV-2 infection. The antiviral mechanism was found to be mediated by the inhibition of a serine protease, TMPRSS2, which results in a blockage of viral entry into target cells. Potential treatments with these serpins would not only reduce the overall viral burden in the patients, but also block the infection at an early time point, reducing the risk for the hyperactive immune response common in patients with severe COVID-19

Type I interferon shapes brain distribution and tropism of tick-borne flavivirus

Viral tropism within the brain and the role(s) of vertebrate immune response to neurotropic flaviviruses infection is largely understudied. We combine multimodal imaging (cm-nm scale) with single nuclei RNA-sequencing to study Langat virus in wildtype and interferon alpha/beta receptor knockout (Ifnar-/-) mice to visualize viral pathogenesis and define molecular mechanisms. Whole brain viral infection is imaged by Optical Projection Tomography coregistered to ex vivo MRI. Infection is limited to grey matter of sensory systems in wildtype mice, but extends into white matter, meninges and choroid plexus in Ifnar-/- mice. Cells in wildtype display strong type I and II IFN responses, likely due to Ifnb expressing astrocytes, infiltration of macrophages and Ifng-expressing CD8+ NK cells, whereas in Ifnar-/-, the absence of this response contributes to a shift in cellular tropism towards non-activated resident microglia. Multimodal imaging-transcriptomics exemplifies a powerful way to characterize mechanisms of viral pathogenesis and tropism

Type I Interferon Autoantibodies Correlate With Cellular Immune Alterations in Severe COVID-19

BackgroundInfection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can lead to severe disease with increased morbidity and mortality among certain risk groups. The presence of autoantibodies against type I interferons (aIFN-Abs) is one mechanism that contributes to severe coronavirus disease 2019 (COVID-19).MethodsThis study aimed to investigate the presence of aIFN-Abs in relation to the soluble proteome, circulating immune cell numbers, and cellular phenotypes, as well as development of adaptive immunity.ResultsaIFN-Abs were more prevalent in critical compared to severe COVID-19 but largely absent in the other viral and bacterial infections studied here. The antibody and T-cell response to SARS-CoV-2 remained largely unaffected by the presence aIFN-Abs. Similarly, the inflammatory response in COVID-19 was comparable in individuals with and without aIFN-Abs. Instead, presence of aIFN-Abs had an impact on cellular immune system composition and skewing of cellular immune pathways.ConclusionsOur data suggest that aIFN-Abs do not significantly influence development of adaptive immunity but covary with alterations in immune cell numbers