Epithelial barrier protection : implications for HIV susceptibility

The majority of HIV infections today occur through sexual HIV transmission. The female genital mucosa offers a barrier against incoming pathogens. Although, studies show that the vaginal microbiome, co-current infections and local inflammation, the use of hormonal contraceptives and microbicides, can weaken this protective lining. In this thesis in situ digital image analysis workflows were developed and used together with protein profiling, to characterize the effects of such factors on the genital mucosal barrier integrity and the immune cells therein. Topically applied microbicides can protect against HIV. In paper I we introduced image analysis as a refined tool for evaluation of microbicide safety. We confirmed that a promising microbicide candidate, Q-GRFT, had no negative effect on the rectal epithelium while causing a small, but probably biologically negligible, increase in CD4+ HIV target cells. We also discovered potential effects of multiple biopsy sampling that should be considered when designing pre-clinical studies. Specific tissue resident immune cells with an effector memory phenotype have a rapid response against re-infections and may be important against HIV infection. In paper II, we observed that HIV infected women had increased levels of CD103-CD8+ tissue resident memory cells compared to uninfected women, and that this may be due to a recent influx of these effector cells that have not yet upregulated the CD103 retention molecule. In paper III we revealed that women taking the hormonal contraceptive DMPA, had a thinner superficial layer of the female genital mucosa. Lack of this protective layer in combination with having more HIV target cells located closer to the vaginal lumen, could contribute to the increased HIV risk in women taking DMPA. In paper IV we discovered that Lactobacillus non-iners dominated women had a more intact epithelium, and Gardnerella dominated women had a different spatial localization of CD4+ cells in the epithelium. Secreted protein profiles from Lactobacillus dominant women had elevated levels of anti-inflammatory and epithelial barrier proteins compared to non-Lactobacillus dominant women. These factors may contribute to reduced HIV risk in Lactobacillus-dominant women. The results of this thesis highlight the benefits of using digital image analysis as a tool for studying spatial and structural changes in the mucosal tissue barrier and the immune cell landscape therein. We showed potential mechanisms in how different factors increase the HIV risk. These findings will support development of interventions aimed to strengthen the mucosal barrier, and thereby reduce transmission of sexually transmitted infections

Habitat selection, elevational shift, and population trend in the Norwegian population of the Eurasian Dotterel (Charadrius morinellus)

Climate change occurs at a faster rate in alpine areas and in northern latitudes compared to other regions. However, we have too little knowledge of the consequences of such changes within alpine areas. Studies regarding this subject have to our knowledge been geographically restricted, mainly to central Europe and the western hemisphere. We conducted a study in Norway during the breeding season regarding the Eurasian Dotterel (Charadrius morinellus). We counted territories, registered site variables, and used historical data to address the population trend and to uncover a potential shift in elevation. We found a significant, negative relationship between the number of territories and the density of sheep. Additionally, different levels of bedrock nutrition influenced the number of territories, where areas with intermediate bedrock richness contained the most territories. The mean elevation of a territory was 1429 m a.s.l. (range: 1221-1656 m a.s.l.) We also found that the dotterel had experienced an elevational shift in Norway from 1970 to 2022, equalling an upward shift of 17.4 m a.s.l. for each decade. If this trend continues, the loss of breeding habitat in our study sites may amount to ca 20% during the next 50 years. However, we found no significant shift in elevation for the period 2008-2022 in the same region. The population trend within our study sites was also stable from 2008 to 2022. Even though we found a stable population trend, a continued shift in the elevation will likely lead to future population declines. A potential management tool that we present here, is to limit the density of grazing sheep in alpine breeding areas. It might also be crucial for management to gather more data regarding population trends in alpine birds and the potential drivers for elevational shifts. Such data could then be used to create mitigating efforts to reduce the effects of these drivers

Ojämlik behandling och diskriminering inom den svenska sjukvården : en undersökning av bidragande faktorers interaktion

The Swedish health care does not live up to the objective set in the legislation about health care, that every person has the equal right to health care according to need. New reports show that both discrimination and unequal treatment among patients occur within the Swedish health care. This study is confined to include only institutions for treatment of people with physical health problems. Previous research show that people with disabilities and chronic disease, low socioeconomic status, no or few family members, and people who differs from the norm concerning gender, culture, age, sexuality etc. gets inferior treatment and care at Swedish hospitals. The aim of this study is to analyse how different factors leading to the occurrence of discrimination and unequal treatment interact with each other. This is done with a literature study combined with qualitative interviews with Swedish doctors. The method is chosen to make it possible to answer the question regarding how the main factors interact with each other and possibly find a mutual source. Other material used for this study is the Swedish legislation and a report from the Swedish discrimination ombudsman, as well as previous findings from Doctor Nina Cavalli-­‐Björkman at the University of Uppsala. The results show that discrimination and unequal treatment depend on several interacting factors joint by the fact that the Swedish healthcare system is underfinanced. The Swedish healthcare supply cannot meet the total demand, which results in discrimination and unequal treatment. Underlying norms and power structures in society result in health personnel making priorities based on deep-­‐rooted norms, which results in unequal treatment and discrimination for certain groups

Northern Crossings

This open access book uses Swedish literature and the Swedish publishing field as recurring examples todescribe and analyse the role of the literary semi-peripheral position in world literature from various perspectives and on meso, micro and macro levels, using both quantitative and qualitative methods. This includes the role of translation in the semi-periphery and the conditions under which literature travels to and from that position. The focus is not on Sweden, as such, but rather on the semi-peripheral transitional space as exemplified by the Swedish case. Consisting of three co-written chapters, this study sheds light on what might be called the semi-peripheral condition or the semi-periphery as an area of transition. As part of the Cosmopolitan and Vernacular Dynamics in World Literatures series, it makes continuous use of the concepts of 'cosmopolitan' and 'vernacular' – or rather, the processual terms, cosmopolitanization and vernacularization – which provide an overall structure to the analysis of literature and literary phenomena. In this way, the authors show that the semi-periphery is an ideal point of departure to further the understanding of world literature, because it is a place where the cosmopolitan (the literary universal) and the vernacular (the rootedness in a particular culture or place) interact in ways that have not yet been thoroughly explored. The open access edition of this book is available under a CC BY-NC-ND 3.0 licence on www.bloomsburycollections.com. Open access was funded by Riksbankens Jubileumsfond

Northern Crossings

This open access book uses Swedish literature and the Swedish publishing field as recurring examples todescribe and analyse the role of the literary semi-peripheral position in world literature from various perspectives and on meso, micro and macro levels, using both quantitative and qualitative methods. This includes the role of translation in the semi-periphery and the conditions under which literature travels to and from that position. The focus is not on Sweden, as such, but rather on the semi-peripheral transitional space as exemplified by the Swedish case. Consisting of three co-written chapters, this study sheds light on what might be called the semi-peripheral condition or the semi-periphery as an area of transition. As part of the Cosmopolitan and Vernacular Dynamics in World Literatures series, it makes continuous use of the concepts of 'cosmopolitan' and 'vernacular' – or rather, the processual terms, cosmopolitanization and vernacularization – which provide an overall structure to the analysis of literature and literary phenomena. In this way, the authors show that the semi-periphery is an ideal point of departure to further the understanding of world literature, because it is a place where the cosmopolitan (the literary universal) and the vernacular (the rootedness in a particular culture or place) interact in ways that have not yet been thoroughly explored. The open access edition of this book is available under a CC BY-NC-ND 3.0 licence on www.bloomsburycollections.com. Open access was funded by Riksbankens Jubileumsfond

Interbase-FRET binding assay for pre-microRNAs

The aberrant expression of microRNAs (miRs) has been linked to several human diseases. A promising approach for targeting these anomalies is the use of small-molecule inhibitors of miR biogenesis. These inhibitors have the potential to (i) dissect miR mechanisms of action, (ii) discover new drug targets, and (iii) function as new therapeutic agents. Here, we designed Forster resonance energy transfer (FRET)-labeled oligoribonucleotides of the precursor of the oncogenic miR-21 (pre-miR-21) and used them together with a set of aminoglycosides to develop an interbase-FRET assay to detect ligand binding to pre-miRs. Our interbase-FRET assay accurately reports structural changes of the RNA oligonucleotide induced by ligand binding. We demonstrate its application in a rapid, qualitative drug candidate screen by assessing the relative binding affinity between 12 aminoglycoside antibiotics and pre-miR-21. Surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) were used to validate our new FRET method, and the accuracy of our FRET assay was shown to be similar to the established techniques. With its advantages over SPR and ITC owing to its high sensitivity, small sample size, straightforward technique and the possibility for high-throughput expansion, we envision that our solution-based method can be applied in pre-miRNA-target binding studies

Application of Optical Biosensors in Small-Molecule Screening Activities

The last two decades have seen remarkable progress and improvements in optical biosensor systems such that those are currently seen as an important and value-adding component of modern drug screening activities. In particular the introduction of microplate-based biosensor systems holds the promise to match the required throughput without compromising on data quality thus representing a sought-after complement to traditional fluidic systems. This article aims to highlight the application of the two most prominent optical biosensor technologies, namely surface plasmon resonance (SPR) and optical waveguide grating (OWG), in small-molecule screening and will present, review and discuss the advantages and disadvantages of different assay formats on these platforms. A particular focus will be on the specific advantages of the inhibition in solution assay (ISA) format in contrast to traditional direct binding assays (DBA). Furthermore we will discuss different application areas for both fluidic as well as plate-based biosensor systems by considering the individual strength of the platforms

Study of disease-relevant polymorphisms in the TLR4 and TLR9 genes: a novel method applied to the analysis of the Portuguese population

Toll-like receptors (TLRs) are cellular receptors that mediate recognition of microbial challenges and the subsequent inflammatory response. Genetic variations within these inflammation-associated genes may alter host-pathogen defence mechanisms affecting susceptibility towards infectious diseases. Taking into account the significance of these genes, we developed a simple and rapid method based in the bi-directional PCR amplification of specific alleles (Bi-PASA) for genotyping known sequence variants in TLR4 (Asp299Gly and Thr399Ile) and TLR9 (T-1237C) genes. This method allows genotype determination in a single reaction and is amenable to large-scale analysis. We used Bi-PASA to characterize the distribution of these polymorphisms in the Portuguese population. A total of 388 randomly selected blood donors of Portuguese origin (203 females and 185 males) were genotyped and allele frequencies were determined. Among the tested individuals, 11.1% and 10.8% were heterozygous for Asp299Gly and Thr399Ile, respectively. In what concerns the T-1237C variation in TLR9, the variant allele was present in 19.4% of the individuals tested. Besides confirming the usefulness of the Bi-PASA in polymorphism analysis, the data presented provide valuable information on TLR polymorphisms in the Portuguese population that can be used to stratify risk patients with increased susceptibility to infection.Carvalho A. was financially supported by a fellowship from Fundação para a Ciência e Tecnologia, Portugal (contract SFRH/BD/11837/2003). This study was supported by Fundação para a Ciência e Tecnologia, Portugal (POCI/SAU-ESP/61080/2004)