Identification of gliadin-binding peptides by phage display

BACKGROUND: Coeliac disease (CD) is a common and complex disorder of the small intestine caused by intolerance to wheat gluten and related edible cereals like barley and rye. Peptides originating from incomplete gliadin digestion activate the lamina propria infiltrating T cells to release proinflammatory cytokines, which in turn cause profound tissue remodelling of the small intestinal wall. There is no cure for CD except refraining from consuming gluten-containing products.RESULTS: Phage from a random oligomer display library were enriched by repeated pannings against immobilised gliadin proteins. Phage from the final panning round were plated, individual plaques picked, incubated with host bacteria, amplified to a population size of 1011 to 1012 and purified. DNA was isolated from 1000 purified phage populations and the region covering the 36 bp oligonucleotide insert from which the displayed peptides were translated, was sequenced. Altogether more than 150 different peptide-encoding sequences were identified, many of which were repeatedly isolated under various experimental conditions. Amplified phage populations, each expressing a single peptide, were tested first in pools and then one by one for their ability to inhibit binding of human anti-gliadin antibodies in ELISA assays. These experiments showed that several of the different peptide-expressing phage tested inhibited the interaction between gliadin and anti-gliadin antibodies. Finally, four different peptide-encoding sequences were selected for further analysis, and the corresponding 12-mer peptides were synthesised in vitro. By ELISA assays it was demonstrated that several of the peptides inhibited the interaction between gliadin molecules and serum anti-gliadin antibodies. Moreover, ELISA competition experiments as well as dot-blot and western blot revealed that the different peptides interacted with different molecular sites of gliadin.CONCLUSIONS: We believe that several of the isolated and characterised gliadin-binding peptides described here could provide valuable tools for researchers in the field of CD by facilitating studies on localisation and uptake of various gliadin peptides in the small intestine. In future work, the potential of these peptides to detoxify gluten will be investigated

Immunophenotype in orofacial granulomatosis with and without Crohn's disease

Objectives: The aim of this investigation was to characterise and compare the inflammatory infiltrates in patients with orofacial granulomatosis solely (OFG-S) and OFG with coexisting Crohn's disease (OFG+CD). Study Design: Biopsy specimens with granulomas were obtained from patients with OFG-S (n=11) and OFG+CD (n=11) and immunostained with antibodies against CD1a, CD3, CD4, CD8, CD11c, CD20, CD68 and mast cell tryptase, followed by quantitative analysis. Results: Analyses of the connective tissue revealed a significantly higher number of CD3- expressing T cells and CD11c-expressing dendritic cells in the connective tissue of patients with OFG-S compared to patients with OFG+CD. Mast cells displayed a high level of activation, although no significant difference was detected when comparing the two groups. Conclusions: The results show a different composition of the inflammatory infiltrate in patients with OFG-S compared to patients with OFG+CD. The present observations support that partly divergent immune mechanisms are involved in these two different subcategories of OFG

pH-dependent structural transitions in cationic ionizable lipid mesophases are critical for lipid nanoparticle function

Lipid nanoparticles (LNPs) are advanced core-shell particles for messenger RNA (mRNA) based therapies that are made of polyethylene glycol (PEG) lipid, distearoylphosphatidylcholine (DSPC), cationic ionizable lipid (CIL), cholesterol (chol), and mRNA. Yet the mechanism of pH-dependent response that is believed to cause endosomal release of LNPs is not well understood. Here, we show that eGFP (enhanced green fluorescent protein) protein expression in the mouse liver mediated by the ionizable lipids DLin-MC3-DMA (MC3), DLin-KC2-DMA (KC2), and DLinDMA (DD) ranks MC3 ≥ KC2 > DD despite similar delivery of mRNA per cell in all cell fractions isolated. We hypothesize that the three CIL-LNPs react differently to pH changes and hence study the structure of CIL/chol bulk phases in water. Using synchrotron X-ray scattering a sequence of ordered CIL/chol mesophases with lowering pH values are observed. These phases show isotropic inverse micellar, cubic Fd3m inverse micellar, inverse hexagonal and bicontinuous cubic Pn3m symmetry. If polyadenylic acid, as mRNA surrogate, is added to CIL/chol, excess lipid coexists with a condensed nucleic acid lipid phase. The next-neighbor distance in the excess phase shows a discontinuity at the Fd3m inverse micellar to inverse hexagonal transition occurring at pH 6 with distinctly larger spacing and hydration for DD vs. MC3 and KC2. In mRNA LNPs, DD showed larger internal spacing, as well as retarded onset and reduced level of DD-LNP-mediated eGFP expression in vitro compared to MC3 and KC2. Our data suggest that the pH-driven Fd3m- transition in bulk phases is a hallmark of CIL-specific differences in mRNA LNP efficacy.publishedVersio

Unga långtidssjuka klienters erfarenheter av sjukhusvistelse och möjlighet till aktivitet

Syftet med examensarbetet är att genom tidigare forskning kartlägga långtidssjuka unga klienters aktivitetsmöjligheter och erfarenheter under sjukhusvistelse. Med hjälp av frågeställningarna undersöks hurudana faktorer som blir viktiga angående aktivitetsmöjligheter för långtidssjuka unga på sjukhus. Som metod används en systematisk litteraturstudie. Som referensram används Value and Meaning of Occupation (ValMO) modellen. Referensramen byggs upp av tre värdetriader: aktivitetstriad, värdetriad, perspektivtriad. Den tar även upp mening och balans i aktivitet. Antalet artiklar som inkluderas efter kvalitetsgranskningen är 15 stycken. Innehållsanalys används för att analysera resultatet. Resultatet sammanfattas i fyra huvudteman; miljöns påverkan av möjlighet till aktivitet, möjlighet till meningsfulla aktiviteter, aktivitetsmönster och erfarenheter av sjukhusvistelsen. Resultatet lyfter fram långtidssjuka ungas positiva och negativa erfarenheter bl.a. angående kommunikation med personal. I resultatet kommer även fram hur miljön kan möjliggöra samt begränsa möjligheter till aktivitet, hur en ung individs aktivitetsmönster ändrar under en sjukhusvistelse och hurudana aktivitetsmöjligheter unga önskar att det fanns vid sjukhus. Examensarbetet är av relevans med tanke på utveckling av en mera möjliggörande sjukhusmiljö samt känsla av välbefinnande och hälsa hos långtidssjuka unga med hjälp av meningsfulla aktiviteter.The aim of the study is to examine how research describes experiences of hospitalization and activity opportunities among chronically ill youth. With the support of the research questions it examines what type of elements becomes important regarding occupational opportunities among hospitalized youth. As a method a systematic literature review was implemented. The Value and Meaning of Occupation (ValMO) model is used as a theoretical framework. The main concepts of the framework are value & meaning of occupation and occupational balance. The number of articles that are included after the quality review are 15. Content analysis is used to analyze the results. The results is divided into four main themes; environmental factors regarding occupational opportunities, possibility to attend meaningful occupations, occupational patterns and experiences of hospitalization. The results highlights hospitalized youths positive and negative experiences of communication with hospital staff, how environmental factors can enable and limit occupational opportunities, how hospitalized youths occupational patterns change during hospitalization and what type of occupational possibilities youths wish hospitals would provide. The work is of relevance regarding development of a more enabling hospital environment and sense of well-being and health of chronically ill youth through meaningful activities

How can you control a relationship? : A study of the management control systems ofinterorganizational relationships between small and medium sized enterprises

Bakgrund: Interorganisatoriska relationer har blivit allt viktigare de senaste åren, företag köper exempelvis in IT-tjänster för att kunna förbättra sin kärnverksamhet. Det är dock vanligt att interorganisatoriska relationer misslyckas, en lösning till problem med dessa relationer är att använda ekonomisk styrning. En typ av relation som är speciellt utsatt för problematiken är relationen mellan små och medelstora företag, vilket är en grupp det saknas forskning och empiriska undersökningar om, trots dess stora inverkan på den svenska ekonomin. Ännu mer problematiskt blir det när dessa företag köper eller säljer tjänster, då hantering av tjänster kräver en större administrativ kunskap som mindre företag generellt inte besitter. Därmed är det av intresse att studera hur relationen mellan små och medelstora företag som hanterar tjänster ska styras på bästa sätt. Syfte: Syftet med studien är att skapa en modell över hur styrningen av interorganisatoriska relationer utformas och används av små och medelstora företag. Genomförande: Studien är en kvalitativ fallstudie med ett abduktivt angreppsätt. Empirisk data har samlats in genom semistrukturerade intervjuer på fem olika små och medelstora företag som köper och säljer tjänster, med syfte att framhäva perspektivet från båda partner i relationen.  Resultat: Studiens resultat visar att det krävs en kombination av formella och informella styrmedel för att ha en fungerande interorganisatorisk relation. Avvägningen av olika styrverktyg beror på olika bakomliggande faktorer som präglar relationen mellan små och medelstora företag men resultatet tydliggör även att tillit och kommunikation har en stor inverkan på behovet av olika slags styrning. Samtidigt har kontraktet en väsentlig del av de studerade interorganisatoriska relationerna. Det är i kontraktet som bland annat finansiella och ickefinansiella målsättningar kan specificeras och kontraktet används som ett underlag för utförandet av tjänsten, uppföljning och avstämning, vilket inte alltid utförs i praktiken. Tilliten som finns mellan parterna i en relation påverkar hur kontraktet sedan används. Studien visar att formella styrmedel och tillit i vissa delar av relationen kan användas som substitut till varandra.Background: The importance of interorganizational relationships has increased in recent years. Companies outsource peripheral components of their operations, such as IT, in order to focus on the main purpose of the company. It is common, however, for these relationships to fail and one solution to the problems arising from interorganizational relationships is the use of management control systems. Relationships between small and medium sized enterprises, in particular, present significant issues. Although they have a great impact on the Swedish economy, there are few scientific studies on them. These companies have an even harder time coping with interorganizational relationships when they are dealing with services, as they require more administrative knowledge than they don’t possess. It is therefore of elevated interest to study the relationship between small and medium sized enterprises that handles services and what type of management control system is best suited for the relationship.  Purpose: The purpose of this study is to create a model of the management control systems of interorganizational relationships between small and medium sized enterprises.  Design: The study is a qualitative case study with an abductive approach. Empirical data has been collected through semi-structured interviews at five different small and medium sized enterprises that sell and buy services in order to elucidate both parties’ perspectives.  Conclusion: The results from the study demonstrate that it is necessary to have a combination of formal and informal management control systems in order to maintain a functioning interorganizational relationship. The balance between different control systems depends on underlying factors arising from the relationship between small and medium sized enterprises. The study also shows that trust and communication has a great impact on the need for different types of control, however, the contract has simultaneously proven to be an essential part of the interorganizational relationships that have been studied. The contract can specify the financial and non-financial aims of the parties and can also be used as a model for how the service should be provided. The contract can also be used for future reviews of the working relationship, although this is not always the case, as the level of trust between the two parties will influence how the contract is actually used. The study shows that formal control systems and trust can be used as substitutes to one another throughout the relationship