Simultaneous Visualization of Both Signaling Cascade Activity and End-Point Gene Expression in Single Cells

We have developed an approach for simultaneous detection of individual endogenous protein modifications and mRNA molecules in single cells in situ. For this purpose we combined two methods previously developed in our lab: in situ proximity ligation assay for the detection of individual protein interactions and -modifications and in situ detection of single mRNA molecules using padlock probes. As proof-of-principle, we demonstrated the utility of the method for simultaneous detection of phosphorylated PDGFRβ and DUSP6/MKP-3 mRNA molecules in individual human fibroblasts upon PDGF-BB stimulation. Further we applied drugs disrupting the PDGFRβ signaling pathway at various sites to show that this combined method can concurrently monitor the molecular effect of the drugs, i.e. inhibition of downstream signaling from the targeted node in the signaling pathway. Due to its ability to detect different types of molecules in single cells in situ the method presented here can contribute to a deeper understanding of cell-to-cell variations and can be applied to e.g. pinpoint effector sites of drugs in a signaling pathway

Genital Chronic Graft-versus-Host Disease in Females: A Cross-Sectional Study

AbstractUsing the National Institutes of Health (NIH) consensus criteria for chronic graft-versus-host disease (cGVHD), we assessed the prevalence, symptoms, and clinical signs of female genital cGVHD in a cross-sectional population-based study. Forty-two women were evaluated at a median of 80 months (range, 13 to 148 months) after undergoing hematopoietic stem cell transplantation (HSCT). Medical history, ongoing medications, and genital signs and symptoms were recorded. Gynecologic examination for the diagnosis and clinical scoring of genital cGVHD was combined with clinical scoring of extragenital cGVHD for the estimation of each patient's global cGVHD score. Biopsy specimens from the genital mucosa were obtained from 38 patients. Genital cGVHD was diagnosed in 22 of 42 patients (52%). Its presence was associated with systemic corticoid steroid treatment of extragenital cGVHD (P = .001), older age (P = .07), and HSCT from a sibling donor (P = .002). Five patients had isolated genital cGVHD. Dryness, pain, smarting pain (P < .05 for all), and dyspareunia (P = .001) were observed more frequently in the women with genital cGVHD. Twelve patients had advanced genital cGVHD (clinical score 3), which was the main factor explaining the high rate (15 of 42) of severe global cGVHD. The rate of genital cGVHD was similar (P = .37) in patients with a follow-up of ≥80 months (10 of 22) and those with a follow-up of <80 months (12 of 20). We found no convincing relationship between clinical diagnosis and histopathological assessment of mucosal biopsy specimens. In our group of women with a long follow-up after HSCT, genital cGVHD was common and in many cases incorrectly diagnosed. Genital cGVHD causes genital symptoms and affects sexual life, and may present without any other cGVHD, warranting early and continuous gynecologic surveillance in all women after HSCT

Functional loss of IKBE leads to NF-KB deregulation in aggressive chronic lymphocytic leukemia

NF-?B is constitutively activated in chronic lymphocytic leukemia (CLL); however, the implicated molecular mechanisms remain largely unknown. Thus, we performed targeted deep sequencing of 18 core complex genes within the NF-?B pathway in a discovery and validation CLL cohort totaling 315 cases. The most frequently mutated gene was NFKBIE (21/315 cases; 7%), which encodes I?B?, a negative regulator of NF-?B in normal B cells. Strikingly, 13 of these cases carried an identical 4-bp frameshift deletion, resulting in a truncated protein. Screening of an additional 377 CLL cases revealed that NFKBIE aberrations predominated in poor-prognostic patients and were associated with inferior outcome. Minor subclones and/or clonal evolution were also observed, thus potentially linking this recurrent event to disease progression. Compared with wild-type patients, NFKBIE-deleted cases showed reduced I?B? protein levels and decreased p65 inhibition, along with increased phosphorylation and nuclear translocation of p65. Considering the central role of B cell receptor (BcR) signaling in CLL pathobiology, it is notable that I?B? loss was enriched in aggressive cases with distinctive stereotyped BcR, likely contributing to their poor prognosis, and leading to an altered response to BcR inhibitors. Because NFKBIE deletions were observed in several other B cell lymphomas, our findings suggest a novel common mechanism of NF-?B deregulation during lymphomagenesis. <br/

Functional loss of IκBε leads to NF-κB deregulation in aggressive chronic lymphocytic leukemia

NF-κB is constitutively activated in chronic lymphocytic leukemia (CLL); however, the implicated molecular mechanisms remain largely unknown. Thus, we performed targeted deep sequencing of 18 core complex genes within the NF-κB pathway in a discovery and validation CLL cohort totaling 315 cases. The most frequently mutated gene was NFKBIE (21/315 cases; 7%), which encodes IκBε, a negative regulator of NF-κB in normal B cells. Strikingly, 13 of these cases carried an identical 4-bp frameshift deletion, resulting in a truncated protein. Screening of an additional 377 CLL cases revealed that NFKBIE aberrations predominated in poor-prognostic patients and were associated with inferior outcome. Minor subclones and/or clonal evolution were also observed, thus potentially linking this recurrent event to disease progression. Compared with wild-type patients, NFKBIE-deleted cases showed reduced IκBε protein levels and decreased p65 inhibition, along with increased phosphorylation and nuclear translocation of p65. Considering the central role of B cell receptor (BcR) signaling in CLL pathobiology, it is notable that IκBε loss was enriched in aggressive cases with distinctive stereotyped BcR, likely contributing to their poor prognosis, and leading to an altered response to BcR inhibitors. Because NFKBIE deletions were observed in several other B cell lymphomas, our findings suggest a novel common mechanism of NF-κB deregulation during lymphomagenesis

The Immemorial Trauma of Emmanuel Levinas : On the relation to death in the philosophy of Emmanuel Levinas

This paper argues that the notion of death is omnipresent and essential for Emmanuel Levinas through all periods of his thought, although not always explicitly thematized. It tries to show this through a close reading of, firstly, his early period as an explicit polemic against Martin Heidegger’s analysis of death in Being and Time; secondly, his middle period, as an implicit polemic against Alexandre Kojèves influential readings of Hegel; and thirdly, his late period, in the light of psychoanalytical theories about trauma, loss, mourning and survival. This paper further argues that this notion of death, explicitly or implicitly present through all periods of Levinas’s thought, can be characterized as a traumatic survival experience in which a refusal to mourn holds sway. Finally, the paper sketches some consequences that this survival experience of death might have for contemporary post-Levinasian philosophy, to be examined in future research

Plan för multifunktionella buffertzoner längs Vramsån påMalörten AB : s jordbruksfastigheter

To create multifunctional buffer strips along a watercourse in an economically sustainable way requires consideration of many different factors. In this plan we have selected pollutant reduction as the main purpose of the buffer strip. We have also strived to achieve minimal income loss due to reduced production, increased biological diversity, good game preservation that leads to higher income from hunting, aesthetical satisfaction and possibility to profit from available subsidies. This is done mainly to make landowners/farmers more interested in creating buffer strips along water courses. As sand is the dominant type of soil in the area, the water course is less affected by surface runoff compared to if the soil would be clay or some other finer texture. Game preservation and subsidies have therefore been the main factors when establishing the width of the strip, because these require wider buffer strips in some cases. Most of the buffer strips have been left as fallow fields and multilayered vegetation systems have been strived for. This means combined tree, bush and grass/herb zones. To be allowed to plant bushes on a fallow field and also receive subsidies we suggest that perennial energy plants should be planted in some parts of the buffer strips. Perennial energy plants have several benefits, such as income when harvested and game preservation. In addition we have suggested planting and self rejuvenation of the local trees and bushes

Entreprenörskap : Starta eget med Drivhusets hjälp

Denna uppsats behandlar ämnet entreprenörskap, svårigheter med att starta eget och vilken hjälp Drivhuset i Karlstad har bidragit med vid nyföretagande. Uppdraget gavs av Drivhuset, en entreprenörskapsorganisation som inriktar sitt arbete mot studenter. Syftet med uppsatsen var att skapa en bild av vilka problem som kan förekomma vid uppstart av ett företag och på vilket sätt Drivhuset kan hjälpa till i dessa situationer. Vi har även undersökt universitetets eventuella inverkan på entreprenörerna. Arbetets inriktning har delats upp på dels en kvalitativ och en kvantitativ analys, där vi skall stå för den kvalitativa delen medan den kvantitativa analysen görs av en annan uppsatsgrupp på universitetet. Datainsamling har skett i form av intervjuer, där vi har haft kontakt med fyra företag som startat sin verksamhet med hjälp av Drivhuset. Intervjuerna har varit relativt öppna och genomförts med hjälp av en delvis strukturerad intervjuguide. De data vi samlat in har vi analyserat i tre steg och sedan sammanställts i vårt resultat. I teoriavsnittet behandlar vi olika syner på vad som är entreprenörskap och dess roll i samhället samt redogör för Drivhuset och andra organisationer i närområdet som arbetar för att främja nyföretagandet. Vi beskriver också kortfattat de vanligaste företagsformerna samt svårigheter med att starta och driva eget företag. De företag vi valt att intervjua är Attityd i Karlstad som är verksamma inom marknadsundersökningsbranschen, Finanslab som arbetar med makro- och fondanalyser, Wellcase som jobbar med produktutveckling samt tagit fram ett CD- och Dvd-fodral och slutligen Propria som tidigare bedrev en internetbaserad bokhandel. I det avslutande kapitlet har vi diskuterat resultatet av intervjuerna och bland annat kommit fram till att antagligen alla kan starta eget företag men en viktig egenskap är att kunna marknadsföra och sälja sin produkt. Det finns vissa förkunskaper som kan vara bra att ha, men de flesta kunskaper kan förvärvas efterhand. Huvudanledningen till att starta eget anses vara friheten att kunna planera sitt arbete på egen hand och ingen av respondenterna menade att det var ett svårt beslut. De mest uppskattade delarna i Drivhusets verksamhet är hjälp med praktiska problem och att ha någon att diskutera sina idéer med. En annan viktig del är förmedlingen av kontakter även om detta inte värdesätts riktigt lika högt. Ett antal idéer till förbättring har även framkommit under intervjuerna. När vi ser till Karlstads Universitets roll kan det konstateras att ingen av respondenterna har fått någon direkt hjälp av dem vid uppstarten av företaget

Hantering av miljömål i nationell plan för transportinfrastruktur

Flera nationella miljömål har bäring på transportsektorn. Tidigare forskning har visat ett gap mellan riksdagens miljömål för transporterna, och de beslut som fattas om konkreta investeringar i vägnät och järnväg. För att nå målen krävs bättre fordon och bränslen samt beteendeförändringar. Men det behövs också en förändrad samhällsplanering som bidrar till minskande behov av transporter, bättre skydd av värdefulla landskap och mer hållbara städer. Denna rapport beskriver hur nationella miljömål tas omhand i den nationella planen för transportinfrastruktur