To Örebro University

BACKGROUND: For unidentified reasons, possibly due to increased immune surveillance, patients with collagenous colitis (CC) and lymphocytic colitis (LC), both forms of microscopic colitis (MC), have lower risk of colorectal cancer than controls and ulcerative colitis (UC) patients. Levels of secreted and cell-bound mediators in MC patients with active disease and in histological remission (HR) compared to UC patients and controls were investigated.

METHODS: Median fluorescence intensity of 54 analytes in colonic biopsies from patients with active CC (n = 21), LC (n = 11), and UC (n = 19); CC-HR (n = 6), LC-HR (n = 9), UC in remission (n = 19), non-diarrhea controls (n = 48), and diarrhea controls (n = 25) was measured using Luminex.

RESULTS: Granzyme B and CCL5 levels were higher in active CC than in UC, whereas CCL4 and CD163 levels were similar in CC and UC, and both groups had higher levels of matrix metalloproteinase (MMP)-1, MMP-3, and tumor necrosis factor receptor II than both control groups. APRIL, BAFF, BCMA, CCL20, CXCL8, chitinase 3-like 1, pentraxin-3, Fas, and IL-33 were higher in UC than MC. Increases in 4-1BB and perforin in MC compared to controls were lower than in UC. Levels of gp130 and IL-6Rα were decreased in MC but increased in UC compared to controls.

CONCLUSIONS: Microscopic colitis patients exhibit increased levels of several analytes, including some associated with CD8+ T lymphocytes, suggesting a different pathogenesis of MC compared to UC. Higher levels of MMP-1 and MMP-3 in CC than LC indicate separate disease entities.

Purpose: To assess vision outcomes, risk of complications and whether age, gender, specific OCT changes and preoperative vision affect the prognosis for postoperative vision development. We seek to clarify the prognosis of LMH after surgery and assess whether complications can be reduced with appropriate management.

Methods: This retrospective study includes consecutive Lamellar Macular Hole (LMH) surgeries at Örebro University Hospital (2013– 2019), re-evaluated using the OCT-based consensus definition. Cases were classified as Epiretinal Membrane Foveoschisis (ERM-F) or LMH. Pre- and postoperative data were collected from medical records. Statistical analyses were performed with support from medical statisticians.

Results: Both the ERM-F and LMH groups showed Highly significant visual development (p-adj = 0.0010 and 0.0012 respectively), with a median visual improvement of 90% and 38%, respectively. Even the older LMH group, in which the majority (81%) exhibited pathological OCT findings, showed a median improvement of 61% and a significant visual improvement (adjusted p = 0.026).

Conclusion: The results indicate good potential for visual improvement in both groups. Even if the LMH group experiences positive vision improvement, these patients would experience further improvement without the aforementioned OCT changes. Early surgery is highly recommended, ideally before photoreceptor damage occurs; if already present, it should be performed as soon as possible to prevent further OCT pathology.

BACKGROUND: In conjunction with post placement in root-filled teeth with periapical healthy conditions, root canal retreatment may be performed to improve the seal of the root canal. Whether root canal retreatment for technical reasons (retreatments in teeth without apical periodontitis (AP)) results in lower frequency of AP is unknown.

OBJECTIVE: To examine whether there is a difference in frequency of AP between roots with root canals retreated for technical reasons, and roots with root canals not retreated before post placement, with a minimum follow-up of 5 years. Also, to examine changes in root filling quality following root canal retreatment for technical reasons.

METHODS: This retrospective study included radiographs of 441 root-filled roots without periapical radiolucencies at baseline, scheduled for post and core treatment. Follow-up data for a minimum of 5 years were available for 305 roots (loss to follow-up 30.8%), 46 of which were retreated for technical reasons. Two calibrated observers assessed root filling sealing quality and length, respectively, and periapical status according to the Periapical Index. The main outcome of the study, AP, was used as the dependent variable and all analyses were performed at root level.

RESULTS: The overall frequency of AP at follow-up was 13.8%. The difference in frequency of AP between retreated (4.3%) and nonretreated (15.4%) root canals was not statistically significant, p = .061. Analyses including only roots with preoperatively inadequate root filling quality showed a statistically significant difference (p = .017) between the two treatment groups (2.4% vs. 22.9%).

CONCLUSIONS: Root canal retreatment for technical reasons before post and core placement significantly reduces the frequency of AP in roots with inadequate root filling quality.

In eusocial insects, the molecular basis of worker reproductivity, including how it changes with eusocial complexity, remains relatively poorly understood. To address this, we used mRNA-seq to isolate genes differentially expressed between ovary-active and ovary-inactive workers in the intermediately eusocial bumblebee Bombus terrestris. By comparisons with data from the advanced eusocial honeybee Apis mellifera, which shows reduced worker reproductivity, we characterized gene expression differences associated with change in worker reproductivity as a function of eusocial complexity. By comparisons with genes associated with queen-worker caste development in B. terrestris larvae, we tested the behavioral-morphological caste homology hypothesis, which proposes co-option of genes influencing reproductive division of labor in adults in morphological caste evolution. We conducted comparisons having isolated genes expressed in B. terrestris worker-laid eggs to remove the potential confound caused by gene expression in eggs. Gene expression differences between the B. terrestris worker phenotypes were mainly in fat body and ovary, not brain. Many genes (86%) more highly expressed in ovary of ovary-active workers were also expressed in worker-laid eggs, confirming egg-expressed genes were potentially confounding. Comparisons across B. terrestris and A. mellifera, and with B. terrestris larvae, returned significant percentage overlaps in differentially expressed genes and/or enriched Gene Ontology terms, suggesting conserved gene functions underpin worker reproductivity as it declines with increasing eusocial complexity and providing support for the behavioral-morphological caste homology hypothesis. Therefore, within bees, both a degree of conserved gene use and gene co-option appear to underlie the molecular basis of worker reproductivity and morphological caste evolution.

The queen-worker caste system of eusocial insects represents a prime example of developmental polyphenism (environmentally-induced phenotypic polymorphism) and is intrinsic to the evolution of advanced eusociality. However, the comparative molecular basis of larval caste determination and subsequent differentiation in the eusocial Hymenoptera remains poorly known. To address this issue within bees, we profiled caste-associated gene expression in female larvae of the intermediately eusocial bumblebee Bombus terrestris. In B. terrestris, female larvae experience a queen-dependent period during which their caste fate as adults is determined followed by a nutrition-sensitive period also potentially affecting caste fate but for which the evidence is weaker. We used mRNA-seq and qRT-PCR validation to isolate genes differentially expressed between each caste pathway in larvae at developmental stages before and after each of these periods. We show that differences in gene expression between caste pathways are small in totipotent larvae, then peak after the queen-dependent period. Relatively few novel (i.e. taxonomically-restricted) genes were differentially expressed between castes, though novel genes were significantly enriched in late-instar larvae in the worker pathway. We compared sets of caste-associated genes in B. terrestris with those reported from the advanced eusocial honeybee, Apis mellifera, and found significant but relatively low levels of overlap of gene lists between the two species. These results suggest both the existence of low numbers of shared toolkit genes and substantial divergence in caste-associated genes between Bombus and the advanced eusocial Apis since their last common eusocial ancestor.

Introduction: Microscopic colitis (MC) is an inflammatory bowel condition with two subtypes, lymphocytic colitis (LC) and collagenous colitis (CC). Unlike patients with ulcerative colitis (UC) and non-inflamed individuals, MC patients have reduced risk of developing colorectal cancer, possibly due to increased immune surveillance in MC patients.

Aim: To examine differences in levels of immunomodulatory molecules, including those involved in immune checkpoint mechanisms, in sera from patients with MC and in colonic biopsies from patients with MC and UC compared with controls.

Methods: Using Luminex, 23 analytes (4-1BB, 4-1BBL, APRIL, BAFF, BTLA, CD27, CD28, CD80, CTLA-4, E-cadherin, Galectin-3, GITR, HVEM, IDO, IL-2Rα, LAG-3, MICA, MICB, PD-1, PD-L1, PD-L2, sCD40L and TIM-3) were studied in serum from patients with active MC (n = 35) and controls (n = 23), and in colonic biopsies from patients with active LC (n = 9), active CC (n = 16) and MC in histological remission (LC n = 6, CC n = 6), active UC (n = 15) and UC in remission (n = 12) and controls (n = 58).

Results: In serum, IDO, PD-1, TIM-3, 4-1BB, CD27, and CD80 were decreased whereas 4-1BBL and IL-2Rα were increased in MC patients compared with controls. In contrast, in biopsies, levels of PD-L2 and 4-1BB were increased in MC and UC patients with active disease. Furthermore, in biopsies from CC and UC but not LC patients with active disease, CTLA-4, PD-1, APRIL, BAFF, and IL-2Rα were increased compared with controls. PD-L1 was increased in CC but not UC or LC patients. CD27 and TIM-3 were decreased in biopsies from MC patients in comparison to controls whereas levels of MICB were decreased in patients with active UC compared with controls.

Conclusions: Compared with non-inflamed controls, levels of soluble and membrane-bound immunomodulatory molecules were systemically and locally altered in MC and UC patients, with most analytes being decreased in serum but enhanced in colonic biopsies. These findings contribute to knowledge about checkpoint molecules and their role as biomarkers in MC and may also contribute to knowledge about possible mechanisms behind the seemingly protective effects of MC against colorectal cancer.

A growth-related QTL on chicken chromosome 1 has previously been shown to influence domestication behaviour in chickens. In this study, we used Red Junglefowl (RJF) and White Leghorn (WL) as well as the intercross between them to investigate whether stress affects the way birds allocate their time between familiar and unfamiliar conspecifics in a social preference test ('social support seeking'), and how this is related to genotype at specific loci within the growth QTL. Red Junglefowl males spent more time with unfamiliar chickens before the stressful event compared to the other birds, whereas all birds except WL males tended to spend less time with unfamiliar ones after stress. A significant QTL locus was found to influence both social preference under undisturbed circumstances and social support seeking. The WL allele at this QTL was associated not only with a preference for unfamiliar individuals but also with a shift towards familiar ones in response to stress (social support seeking). A second, suggestive QTL also affected social support seeking, but in the opposite direction; the WL allele was associated with increased time spent with unfamiliar individuals. The region contains several possible candidate genes, and gene expression analysis of a number of them showed differential expression between RJF and WL of AVPR2 (receptor for vasotocin), and possibly AVPR1a (another vasotocin receptor) and NRCAM (involved in neural development) in the lower frontal lobes of the brains of RJF and WL animals. These three genes continue to be interesting candidates for the observed behavioural effects. 

Studying animal domestication offers an opportunity to understand the mechanisms of evolution. Domestication is associated with a change in selection pressures; selection for production traits is introduced, and animals are faced with larger and denser social groups. It is not unexpected then that domestication produces a simultaneous change in a number of traits, both physiological and behavioural. This correlated change in traits, e.g. egg production and social behaviour has been termed the “domestic phenotype”. However, it has been shown that selection for one trait alone among the many associated with the domestic phenotype can lead to simultaneous changes in others. This may be a result of such traits being inherited together because of pleiotropy or close linkage of several genes affecting different traits. A chicken growth QTL has previously been found in an intercross between White Leghorn layers (WL) and their main wild ancestor, the red junglefowl (RJF). This QTL has also been found to influence explorative and social behaviours. This thesis aims to characterize this QTL further with respect to social and emotional behaviours, and tries to clarify whether pleiotropy or linkage is responsible for the many observed effects. This is done using behavioural phenotyping, genetic marker genotyping, QTL- and gene expression analysis of an intercross line between RJF and WL, and to some extent of the parental RJF and WL lines themselves. The results show that domestication in these chickens has led to increased social tolerance to unfamiliar conspecifics and a tendency to a decrease in the propensity of chickens to explore the environment, and that these effects are partly explained by the previously described growth QTL. The results also indicate that close linkage of genes, rather than pleiotropy, may be responsible for the multiple effect of the QTL, as different traits to some extent seem to be influenced by different areas within the larger QTL region. This information, in combination with that of other studies and with existing and upcoming genetic research techniques, may be used in the design of future breeding programs that take animal behaviour and welfare as well as production traits into account. Findings like these may also be of use in directing research in human psychiatric genetics.

Domestication of animals, regardless of species, is often accompanied by simultaneous changes in several physiological and behavioral traits (e.g. growth rate and fearfulness). In this study we compared the social behavior and emotional reactivity, as measured in a battery of behavioral tests, of two groups of chickens selected from a common genetic background, an advanced intercross line between the ancestral red junglefowl (“RJF”) and the domesticated White Leghorn layer (“WL”). The birds were selected for homozygosity for alternative alleles at one locus (a microsatellite marker), centrally positioned in a previously identified pleiotropic growth QTL on chromosome 1, closely linked to one major candidate gene (AVPR1a) for certain aspects of social behavior. Birds homozygous for the WL allele (“WL genotype”) had a modified pattern of social and emotional reactions than birds homozygous for the RJF allele (“RJF genotype”), shown by different scores in a principal components analysis. These results suggest that the growth QTL affects a number of domestication related behavioral traits, and may have been a primary target of selection during domestication. The QTL contains a multitude of genes, several of which have been linked to social behavior (for example the vasotocin receptor AVPR1a targeted in this experiment). Future studies aimed at making a higher resolution genotypic characterization of the QTL should give more information about which of these genes may be considered the strongest candidates for bringing about the behavioral changes associated with animal domestication.

Domestication is an evolutionary process in which animals become adapted to a life in close proximity to humans. There are typically specific selection pressures associated with this, including living in larger social groups than their wild ancestors. We hypothesized that the genotype at a major growth QTL could affect aspects of social behavior in chickens as well. We performed social behavior tests in red junglefowl (RJF) and White Leghorn (WL) chickens and in chickens from a selected advanced intercross line (SAIL) between RJF and WL, selected for different genotypes at a microsatellite marker locus within the QTL region. Four-week-old pure WL inspected strangers significantly more than pure RJF. Male 4-wk-old SAIL birds, homozygous for the WL allele at the marker locus, differed from those with RJF alleles in a similar way as the pure WL differed from RJF. Furthermore, 155- to 170-d-old male SAIL birds homozygous for the WL allele at the marker locus were less aggressive to unfamiliar conspecifics in a dominance test. The results suggest that domestication has caused changes in social behavior, which, in males, may partly depend on variations in the genotype at the growth QTL where the avian homolog of the arginine vasopressin receptor 1a (AVPR1a) is located. This gene is therefore one of several putative candidate genes for future research.