Mucins and Pathogenic Mucin-Like Molecules Are Immunomodulators During Infection and Targets for Diagnostics and Vaccines

Mucins and mucin-like molecules are highly O-glycosylated proteins present on the cell surface of mammals and other organisms. These glycoproteins are highly diverse in the apoprotein and glycan cores and play a central role in many biological processes and diseases. Mucins are the most abundant macromolecules in mucus and are responsible for its biochemical and biophysical properties. Mucin-like molecules cover various protozoan parasites, fungi and viruses. In humans, modifications in mucin glycosylation are associated with tumors in epithelial tissue. These modifications allow the distinction between normal and abnormal cell conditions and represent important targets for vaccine development against some cancers. Mucins and mucin-like molecules derived from pathogens are potential diagnostic markers and targets for therapeutic agents. In this review, we summarize the distribution, structure, role as immunomodulators, and the correlation of human mucins with diseases and perform a comparative analysis of mucins with mucin-like molecules present in human pathogens. Furthermore, we review the methods to produce pathogenic and human mucins using chemical synthesis and expression systems. Finally, we present applications of mucin-like molecules in diagnosis and prevention of relevant human diseases

Motivation to study among students in the individual programme : Emphasizing the importance of the parents

Denna studie utgörs av en kvalitativ undersökning där sex stycken gymnasieelever intervjuas. Eleverna har tidigare gått på det individuella programmet och därefter påbörjat ett nationellt gymnasieprogram. Syftet med studien är att ta reda på vad som motiverar elever på det individuella programmet med fokus på hur elever uppfattar föräldrars möjlighet att påverka studiemotivationen. Respondenterna beskriver vad motivation betyder för dem och vad som motiverar dem i deras studier. Därefter läggs fokus på hur respondenterna upplever att deras föräldrar hjälpt dem i studierna samt föräldrarnas betydelse för studiemotivationen. I resultatet framgår att studiemotivationen påverkas av bland annat elevens viljestyrka, målsättningar, känslor och av andra människor. Resultatet beskriver både inre och yttre motivationsfaktorer. Hur mycket föräldrarna hjälper sina barn varierar, men de elever som får läxhjälp eller uppmuntran i studierna upplever att föräldrarna motiverar dem i studierna. Studiens resultat visar även att en god relation mellan eleven och föräldrarna bidrar både till högre studiemotivation hos eleven och till ökad praktisk hjälp från föräldrarna i elevens studier. För elever med mindre god relation till föräldrarna kan istället andra personer bli betydelsefulla för elevens studiemotivation.

Local release of lithium from sol-gel coated orthopaedic screws : an in vitro and in vivo study

  In orthopaedic practice, fractures are usually stabilised with metal screws or rods. This is done in order to keep the fracture parts in place during the rather slow healing process. The healing time can potentially be reduced by local- or systemic treatment with different bone promoting drugs. In later years, lithium, otherwise used to treat bipolar disease, has shown promise to be such a drug.   The aim of this master thesis was to find a way to coat metal bone screws with lithium and to characterise the coating. The coating was to be designed in such a way that it could release lithium to the surrounding bone tissue.   Lithium chloride was incorporated into a titanate sol-gel and attached to silicon wafers and stainless steel screws by dip coating. Wafers were used for initial in vitro studies of how lithium changed coating characteristics. This was studied using ellipsometry, AFM and SEM. Lithium is most probably physisorbed and not incorporated into the network building up the sol-gel. Coating structure is changed as more lithium is incorporated. For large amounts of lithium, the nanoparticles normally formed when curing the sol-gel are inhibited. One effect of this is reduced bioactivity, seen as a reduced ability for calcium phosphate crystals to nucleate on the coating when immersed in simulated body fluid. Lithium release was investigated using AAS. Lithium is released from the coating, showing a burst effect. By changing the number of coating layers used, the release profile can be partly altered. The coating was also applied to screws, showing good attachment, and the lithium release profile was similar to the one seen from wafers. Finally, a screw model was used in rats to assess the effect of local lithium treatment from screws and systemic lithium treatment on fracture healing. In the model, a screw was inserted in tibia, mimicking a fracture. When the bone around the screw was healed, a pullout test was performed, giving information about the strength of the bone surrounding the screw. No significant difference could be found for either local- or systemic lithium treatment compared to control. However, when evaluating the strength of intact bone in a similar way, a positive effect of systemic lithium treatment could be seen. Therefore, it is still likely that lithium has a positive effect on bone and further studies are needed to fully evaluate its role in fracture healing.  Vid behandling av benbrott stabiliseras vanligtvis frakturen internt med metallskruvar och metallstavar. Detta görs för att hålla brottbitarna på plats under den relativt långsamma läkprocessen. Det är möjligt att minska tiden för frakturläkning genom att lokalt eller systemiskt behandla med olika läkemedel som främjar bentillväxt. På senare år har det presenterats bevis för att litium, som annars används som psykofarmaka, fungerar som ett sådant läkemedel.   Syftet med detta examensarbete var att hitta en metod för att fästa litium på benimplantat. Litium skulle fästas på ett sådant sätt att frisläppning till omgivande vävnad blev möjlig.   Litiumklorid inkorporerades i en titanat-solgel och lager av detta lades på kiselytor och rostfria skruvar genom s.k. ”dip-coating”. Kiselytorna användes för initiala in vitro-studier av hur litium ändrade beläggningens egenskaper. Litium sitter antagligen fast på ytan av det tredimensionella nätverk som utgör solgelen, istället för att sitta inbundet i nätverket. Lagerstrukturen ändras ju mer litium som inkorporeras och vid stora mängder skapas inte de nanopartiklar som vanligtvis finns i en solgel-baserad beläggning. En följd av detta är reducerad bioaktivitet för beläggningen, dvs. en minskad förmåga för kalciumfosfatkristaller att bildas på ytan. Litium frisläpps från beläggningen, dock sker denna frisläppning snabbt. Genom att belägga ytan med flera lager av solgel kan frisläppningskinetiken delvis ändras. Solgelen kunde också med god vidhäftning appliceras på skruvar och frisläppningskinetiken från en skruv är liknande den från en kiselyta. Slutligen användes en skruvmodell i råtta för att undersöka vilken effekt lokal respektive systemisk litiumbehandling har på frakturläkning. I modellen efterliknas ett benbrott genom att en skruv sätts in i skenbenet.  När benvävnaden runt skruven har läkt görs ett utdragstest på skruven vilket ger information om benets styrka. Ingen signifikant skillnad i skruvens utdragskraft kunde ses mellan de båda försöksgrupperna och kontrollgruppen. Däremot hade gruppen som fick systemisk litiumbehandling fått starkare ben totalt, vilket indikerar att litium har effekt på intakt ben. På grund av dessa resultat finns det fortfarande skäl att tro att litium har en positiv påverkan på ben, varför dess effekt på frakturläkning bör undersökas ytterligare.

En utredning av kommunala markanvisningspolicyer : Påverkan av lag 2014:899

Det är sedan tidigare tydligt att det funnits problem och delade åsikter kring arbetet med markanvisningar innan lag 2014:899 trädde i kraft. Arbetets syfte var att undersöka om arbetet har förändrats efter att lag 2014:899 trädde i kraft. Målet med arbetet är att utreda de valda kommunernas markanvisningspolicyer och deras erfarenheter av arbetet med dessa. Förbättringsåtgärder föreslås efter att ha belyst svagheter i de markanvisningspolicyer som undersöktes, med utgångspunkt i lag 2014:899 samt punkter specificerade av Caesar et al. (2013).  Ett dokument med förslag till vad en markanvisningspolicy bör innehålla har tagits fram. Dokumentet kan underlätta kommuners arbete med markanvisningspolicyer i fortsättningen

Horse-related trauma in children and adults during a two year period

Introduction: Horse riding, with almost 200,000 participants, is the eighth most popular sport in Sweden. Severe injuries can occur with horse riding accidents which is well documented. This study was undertaken to investigate if injuries associated with horse riding are common, which type of injuries occur, what mechanisms are involved and to estimate the costs to the society. Material and methods: All patients attending the emergency department at Link ping University Hospital, during the years 2003-2004, due to horse related trauma were prospectively recorded. The patients were divided into two groups according to age, 147 children and 141 adults. The medical records were retrospectively scrutinized. Results: The most common mechanism of injury was falling from the horse. Most commonly, minor sprains and soft tissue injuries were seen, but also minor head injuries and fractures, mainly located in the upper limb. In total 26 adults and 37 children were admitted. Of these 63 patients 19 were considered having a serious injury. In total, four patients needed treatment in intensive care units. The total cost in each group was 200,000 Euro/year. Conclusion: Horse riding is a sport with well known risks. Our results corresponds to the literature, however we have not observed the same incidence of serious injuries. In contrast we find these to be fairly uncommon. The injuries are mainly minor, with a small risk of long term morbidity. Over time regulations and safety equipment seem to have decreased the number of serious accidents

Acoustic monitoring of changes in well-defined hyaluronan layers exposed to chondrocytes.

The interaction of human-derived chondrocytes and thin hyaluronan layers was studied using the quartz crystal microbalance with dissipation (QCM-D) technique combined with light microscopy. This approach allowed unique real-time monitoring of the interface between the cells and the sensor surface. Our results suggest that the hyaluronan layer is rapidly degraded by chondrocytes.peerReviewe

Immobilization of chondroitin sulfate to lipid membranes and its interactions with ECM proteins

Glycosaminoglycans (GAGs) in the extracellular matrix (ECM) have multiple functions in tissues including providing support, mediating cell division and differentiation, and taking part in important interactions with proteins, e.g. growth factors. Studying GAG related interactions is inherently difficult and requires suitable interaction platforms. We show two strategies to covalently couple the GAG chondroitin sulfate (CS) to supported lipid bilayers (SLBs), either by (a) activating carboxy-functionalized phospholipids in the lipid bilayer, followed by the addition of hydrazide-functionalized CS, or by (b) activating naturally occurring carboxyl groups on CS prior to addition to an amino-functionalized SLB. Bilayer formation and subsequent immobilization was followed in real-time using the Quartz Crystal Microbalance with Dissipation monitoring, a technique that provides unique information when studying highly hydrated molecular films. The two strategies yielded thin CS films (in the nanometer range) with similar viscoelastic properties. Fluidity of the lipid bilayer was retained when CS was coupled. The application of the CS interaction platform was exemplified for type I collagen and the bone inducing growth factor bone morphogenetic protein-2 (BMP-2). The addition of collagen to immoblized CS resulted in soft layers whereas layers formed by addition of BMP-2 were denser, independent on the immobilization strategy used

Study on multilayer structures prepared from heparin and semi-synthetic cellulose sulfates as polyanions and their influence on cellular response

Multilayer coatings of polycationic chitosan paired with polyanionic semi-synthetic cellulose sulfates or heparin were prepared by the layer-by-layer method. Two different cellulose sulfates (CS) with high (CS2.6) and intermediate (CS1.6) sulfation degree were prepared by sulfation of cellulose. Multilayers were fabricated at pH 4 and the resulting films were characterized by several methods. The multilayer 'optical' mass, measured by surface plasmon resonance, showed little differences in the total mass adsorbed irrespective of which polyanion was used. In contrast, 'acoustic' mass, calculated from quartz crystal micro balance with dissipation monitoring, showed the lowest mass and dissipation values for CS2.6 (highest sulfation degree) multilayers indicating formation of stiffer layers compared to heparin and CS1.6 layers which led to higher mass and dissipation values. Water contact angle and zeta potential measurements indicated formation of more distinct layers with using heparin as polyanion, while use of CS1.6 and CS2.6 resulted into more fuzzy intermingled multilayers. CS1.6 multilayers significantly supported adhesion and growth of C2C12 cells where as only few cells attached and started to spread initially on CS2.6 layers but favoured long term cell growth. Contrastingly cells adhered and grew poorly on to the layers of heparin. This present study shows that cellulose sulfates are attractive candidates for multilayer formation as potential substratum for controlled cell adhesion. Since a peculiar interaction of cellulose sulfates with growth factors was found during previous studies, immobilization of cellulose sulfate in multilayer systems might be of great interest for tissue engineering applications

Probing the biofunctionality of biotinylated hyaluronan and chondroitin sulfate by hyaluronidase degradation and aggrecan interaction

Available online 5 June 2013Molecular interactions involving glycosaminoglycans (GAGs) are important for biological processes in the extracellular matrix (ECM) and at cell surfaces, and also in biotechnological applications. Enzymes in the ECM constantly modulate the molecular structure and the amount of GAGs in our tissues. Specifically, the changeable sulfation patterns of many GAGs are expected to be important in interactions with proteins. Biotinylation is a convenient method for immobilizing molecules to surfaces. When studying interactions at the molecular, cell and tissue level, the native properties of the immobilized molecule, i.e. its biofunctionality, need to be retained upon immobilization. Here, the GAGs hyaluronan (HA) and chondroitin sulfate (CS), and synthetically sulfated derivatives of the two, were immobilized using biotin-streptavidin binding. The degree of biotinylation and the placement of biotin groups (end-on/side-on) were varied. The introduction of biotin groups could have unwanted effects on the studied molecule, but this aspect that is not always straightforward to evaluate. Hyaluronidase, an enzyme that degrades HA and CS in the ECM, was investigated as a probe to evaluate the biofunctionality of the immobilized GAGs, using both quartz crystal microbalance and high-performance liquid chromatography. Our results showed that end-on biotinylated HA was efficiently degraded by hyaluronidase, whereas already a low degree of side-on biotinylation destroyed the degrading ability of the enzyme. Synthetically introduced sulfate groups also had this effect. Hence hyaluronidase degradation is a cheap and easy way to investigate how molecular function is influenced by the introduced functional groups. Binding experiments with the proteoglycan aggrecan emphasized the influence of protein size and surface orientation of the GAGs for in-depth studies of GAG behavior.The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/20072013) under grant agreement no NMP4-SL2009-229292 ("Find & Bind"). J.B., S.M and M.S. further acknowledge financial support by the Deutsche Forschungsgesellschft, DFG (TRR 67), and S.S. acknowledges financial support provided by the Swedish Research Council (Linnaeus program SUPRA). Gabriel Ohlsson is acknowledged for help with sputter-coating. Anton Paar, Austria, is acknowledged for providing the SurPASS instrument used for the surface zeta-potential measurements