Bc. Klára Horáčková DIPLOMA THESIS Mechanisms of immune dysregulation leading to inflammatory bowel disease Abstract Inflammatory bowel disease (IBD) is a complex disorder characterized by chronic inflammation of the gastrointestinal tract. Classical IBD is a multifactorial disease with adulthood or later-childhood onset. However, children with very early onset IBD (VEO-IBD, before 6 years of age) are a specific cohort, whose pathology can be caused by severe genetic defects in genes connected to immune homeostasis in the gut. We aimed to identify the causal genetic variants in 20 pediatric patients diagnosed with IBD (age of onset from 3 to 154 months) using whole exome sequencing (WES). We evaluated several bioinformatical approaches for WES data analysis. This included a comparison of two methods of variant identification using VarScan2 or GATK4-based tools. Furthermore, we compared 4 gene lists ("virtual panels") for variant filtering, one of which was compiled purposefully for this thesis. We identified and validated via segregation analysis 5 causal variants in 4 genes (DUOX2 compound heterozygote, FOXP3, NLRP3 and NOD2) accounting for 20 % of the cohort. NOD2 (p.A755V) variant has already been reported in IBD cases, while DUOX2 (p.R1216W + p.A1131T), FOXP3 (p.H400L) and NLRP3 (p.V200M) were newly...

This diploma thesis is focused on patients who are undergoing surgery - intestinal resection. In a surgical patient, malnutrition is a frequent and significant negative factor that affects postoperative morbidity and lethality. Proper nutritional care has an irreplaceable role in the prevention and treatment of malnutrition. Nutritional elements are part of the perioperative care of the modern ERAS concept, which leads to reduction in the frequency of postoperative complications and a shortening of the hospitalization duration. The concept emphasizes early physical rehabilitation and shortening of the fasting period around the time of the operation, which leads to a significant saving of body protein. The theoretical part describes particular diseases that lead to surgical treatment. Special attention is paid to idiopathic intestinal inflammations and colorectal cancer. It also includes current recommendations of professional companies. The primary objective of the practical part is to compare two groups of patients who underwent intestinal resection. To find out whether targeted nutritional intervention (before, during and after surgery) has an impact on postoperative convalescence, the frequency of complications and the overall length of hospitalization. The aim was also to improve the quality of...

Background: Inflammatory Bowel Disease (IBD) is a chronic, relapsing, autoimmune disease, affecting one in every 150 Canadians. Failure to induce remission of IBD with pharmacotherapy can necessitate surgical interventions, such as the creation of an ostomy. Ostomy surgery can help manage severe IBD and thus improve quality of life; however, individuals living with IBD report the possibility of ostomy surgery as a top concern, which can lead them to refuse or delay this decision until the disease becomes life threatening. Research Objective: The aim of this study is to understand what factors influence the decision to have ostomy surgery in young women with IBD, how the perception of the surgery compares to the reality of living with an ostomy, and the role healthcare professionals play in this decision. Methods: Nine participants who (1) identify as female, (2) are between the ages of 19 and 30, and (3) are currently living with an ostomy to treat IBD were recruited for this study. Additionally, seven healthcare professionals who work with IBD patients were recruited. Participants were invited for an individual, semi-structured interview. Findings: Young adult women living with an ostomy to treat their IBD reflected on their initial fears and concerns about undergoing surgery. Due to the severity of their illness, the majority of participants had requested surgery after having some time to adjust to the idea. This request, however, was often met with resistance or obstacles in the healthcare system. Healthcare professionals share mixed perceptions of ostomy surgery, with some viewing it as a last resort and others perceiving it as a treatment option. Once surgery had been performed, young adult women describe some challenges adjusting to life with an ostomy; but the majority report experiencing an overall improvement in quality of life. Conclusion: Understanding the perceptions that influence how young women perceive ostomies prior to versus after surgery will help identify the factors that influence the decision-making process for ostomy surgery, such as gender, age and stigma. Challenging current beliefs and assumptions may allow more supportive conversations between healthcare professionals and patients and provide insight on the actual lived experience of young women living with an ostomy. / Graduate / 2023-01-13

Bakgrund: Inflammatorisk tarmsjukdom (IBD) inkluderar Ulcerös kolit och Crohns sjukdom som kännetecknas av återkommande inflammation i tarmen. Etiologin för sjukdomarna är fortfarande okänd och forskning visar en ökning, framför allt i Europa. För att främja hälsa trots en livslång sjukdom behöver individen ta ansvar för att anpassa sin sjukdom genom egenvård, detta för att förebygga exacerbation. Syfte: Syftet med studien var att sammanställa personers upplevelser av att anpassa sitt liv utifrån sin kroniska inflammatoriska tarmsjukdom.  Metod: En beskrivande litteraturstudie baserat på 12 kvalitativa studier. Databaserna som användes vid artikelsökningarna var PubMed och CINAHL. Sökbegränsningarna var engelska, människor och 10 år.  Resultat: En återkommande upplevelse bland personerna med IBD var att livet blev begränsat både gällde skola- och arbetsliv, det sociala livet och kosten. För att anpassa sitt liv med en IBD-sjukdom upplevde personerna att kunskap och stöd från andra var betydande. För att hålla sjukdomen i remission upplevdes behandlingar och kostanpassningar vara en viktig fördel. Upplevelsen av sig själv förändrades där en känsla av att inte vara normal var vanligt, att acceptera och hitta mening i livet var en viktig anpassning och resulterade i ett bättre mentalt välbefinnande. Slutsats: Sjukdomen upplevdes skapa begränsningar i det dagliga livet och ändrade självbilden hos personerna. Att acceptera sitt liv med IBD ansågs vara en viktig anpassning för att skapa möjlighet till att leva sitt liv utan dessa upplevda begränsningar. Enligt författarna kommer resultatet ge sjuksköterskor en ökad förståelse gällande omvårdnaden för personer med en IBD-sjukdom samt möjliggöra en fungerande egenvård för dessa personer. / Background: Inflammatory bowel disease (IBD) include Ulcerative Colitis and Crohn’s disease who are characterized by recurrent inflammation in the bowel. The etiology of the diseases is still unknown, and research shows that it’s increasing, especially in Europe. To promote health despite a lifelong illness, the individual needs to take responsibility for their self-care to keep the disease in check. Aim: The aim of this study was to describe persons experience of adapting their life with their inflammatory bowel disease. Method: A descriptive literature study based on 12 qualitative studies. The databases used were PubMed and CINAHL. The study's limitations were English, humans, and 10 years.  Results: A recurring experience among people with IBD was that life was limited in school, work, social life and diet. Knowledge and support from others were important to adapt to a life with IBD. To keep the disease in remission, treatments and dietary adjustments were perceived as an important advantage. The experience of yourself changed, where a feeling of not being normal was common. Patients who were able to accept their disease, and who also transcended to find a higher purpose, ultimately improved their mental health.   Conclusion: The disease creates limitations in daily life and changes the self-image of people. Accepting a life with IBD was an important adjustment to create the opportunity to live one's life without these perceived limitations. According to the authors, the results will give nurses an increased understanding of caregiving and improved self-care for people with IBD.

Cancer treatments including chemotherapy and radiotherapy treat cancer by targeting rapidly dividing cells. Although these forms of treatment damage rapidly dividing cancer cells, they are also toxic to the cells of the gastrointestinal tract, leading to inflammation of the mucosal layer (mucositis) and causing nausea, vomiting, diarrhea, and abdominal pain. Improvement in symptoms may allow patients to have better performance status permitting ongoing treatment and possibly a better prognosis. This article describes the pathophysiology of chemotherapy-induced mucositis and includes 3 case reports of treatment of mucositis with serum bovine immunoglobulin.

SAMMANFATTNING Livet förändras både fysiskt och emotionellt när individen får en diagnos med en kronisk sjukdom. Den sjuke tvingas att anpassa och acceptera att livet förändras. Hur livet med sjukdomen upplevs och hur personen anpassar sig, är individuellt. Även anhöriga och vänner runt den sjuke drabbas. Vården kan bli en viktig del i den sjukes livsvärld och det är viktigt att lyfta fram hur den som är sjuk upplever sin sjukdom och sin vardag. Syfte: Syftet var att beskriva personers upplevelser av att leva med kronisk inflammatorisk tarmsjukdom och hur sjukdomen inverkar på deras vardag. Metod: En litteraturöversikt valdes som metod. Resultatet består av 16 vetenskapliga artiklar, publicerade mellan 2005-2016 vilka passade till studiens syfte och frågeställning. I analysen hittade författarna meningsenheter som omvandlades till koder, koderna delades in i sub-kategorier och dessa delades in i huvudkategorier. Resultat: Tre huvudkategorier kunde utläsas efter analysarbetet; begränsningar i vardagen, emotionell påverkan och kontakter med vården. Begränsningar i vardagen beskrevs vara okontrollerad tarm, fysiska symtom, begränsning av att inte kunna medverka vid aktiviteter, arbetet och nutritionen. Emotionell påverkan var kopplad till maktlöshet, oro, skuld och bristande förståelse. Kontakter med vården beskrivs utifrån vårdens stöd och behandlingar. Slutsats: Stora delar av livet påverkas för personer som lever med kronisk inflammatorisk tarmsjukdom. Varje person upplever sin situation individuellt och har olika erfarenheter och tillgång till stöd. För att skapa kontroll av sin sjukdom krävs det att flera faktorer samspelar med varandra. Det gäller att prova sig fram till vad som fungerar bäst för varje individ. Här är det viktigt att vården är lyhörd och anpassar vården utefter varje individs individuella behov. / ABSTRACT Life changes both physically and emotionally when an individual is diagnosed with a chronical illness. The person is forced to adapt and accept that life changes. How life as ill is perceived individually and how the individual adapts to a new situation differs. Even relatives and friends around the patient suffer. The care can be an important part of the patient’s life-world and it is important to highlight how the patient experience their illness and their daily lives. Aim: The aim was to describe peoples’ experiences of living with chronic inflammatory bowel disease and how the disease affects their daily lives. Method: A literature review was chosen as method. The result consists of 16 scientific articles, published between 2005-2016 which were suited to the purpose of the study and research question. In the analysis aouther found meaning units that were converted into codes, the codes were divided into sub-categories and these were divided into major categories. Results: Three main categories could be seen after the analysis; limitations in everyday life, emotional impact and contacts with healthcare. Limitations in everyday life were described to be uncontrollable bowel, physical symptoms, and limitation of not being able to participate in activities, work and nutrition. Emotional impact was linked to powerlessness, anxiety, guilt and lack of understanding. Contacts with healthcare were based on healthcare support and treatment. Conclusion: Big parts of life is affected for people living with chronic inflammatory bowel disease. Every individual experience their situation differently and have different experiences and access to support. In order to control their illness they require several factors that interact with each other. It’s up to every individual to try and find out what works best for them. Here it’s important that the healthcare personal is responsive and adapt healthcare along each person's individual needs.

Bakgrund: IBD är en kronisk inflammatorisk tarmsjukdom som innefattar sjukdomarna ulcerös kolit och Crohns sjukdom. Sjukdomen har ökat kraftigt i förekomst de senaste tre decennierna, även i Sverige. En person som har en kronisk sjukdom får en påverkad tillvaro och behöver stöd utöver bara kunskap om sjukdomen för att kunna hantera den. Professionell attityd och empati är viktiga aspekter för att skapa ömsesidigt förtroende och respektfull vård för personer med IBD. Syfte: Litteraturstudiens syfte var att belysa personers (≥18 år) upplevelser av att leva med inflammatorisk tarmsjukdom (IBD). Metod: En litteraturstudie med kvalitativ ansats, som omfattade 11 vetenskapliga artiklar med kvalitativ metodik, genomfördes. Databaserna CINAHL och PubMed användes för databassökning. Analysen genomfördes som en stegvis process inspirerad av innehållsanalys. Resultat: Personernas upplevelse av att leva med IBD sammanställdes i två kategorier; ‘Upplevelser ur ett inre perspektiv’ och ‘Upplevelser ur ett relationellt perspektiv’. Kategorierna delades därefter in i nio subkategorier; ‘Oförutsägbarhet och kontrollförlust’, ‘Oro för framtiden’, ‘Kroppsbild och stigma’, ‘Fysiska påfrestningar’, ‘Optimism’, ‘Sociala situationer’, ‘Familj och partner’, ‘Arbete samt ‘Sjukvård’. Konklusion: Personer med IBD upplever både fysiska och psykiska påfrestningar som kräver en helhetssyn i omvårdnaden. En individanpassad omvårdnad är avgörande för att upprätthålla förtroendet för vården och främja positiva resultat för personens behandling och hälsa. Sjuksköterskor bör vara lyhörda för de individuella behoven och ta hänsyn till de psykiska påfrestningar som sjukdomen kan bidra till. Det är också viktigt att skapa en trygg miljö för personerna, då sjuksköterskan kan fungera som en stödjande faktor vid sociala problem. Litteraturstudien visar därmed att omvårdnad spelar en betydande och värdefull roll i personers upplevelser av att leva med IBD. / Background: IBD is a chronic inflammatory bowel disease that includes ulcerative colitis and Crohn disease. It has significantly increased in incidence over the past three decades, in Sweden as well. A person with chronic illness experiences an impacted life and requires support beyond just knowledge of the disease in order to be able to manage it. Professional attitude and empathy are important aspects to create mutual trust and respectful care for people with IBD. Aim: The aim was to illuminate people’s (≥18 years) experiences of living with inflammatory bowel disease (IBD). Method: A literature review with a qualitative approach, which included 11 scientific articles with qualitative methodology. The databases CINAHL and PubMed were used for the database searching. The analysis was conducted as a stepwise process inspired by content analysis. Results: The people’s experience of living with IBD was compiled into two categories; ‘Experiences from an inner perspective’ & ‘Experiences from a relational perspective’. The categories were then divided into 9 subcategories; ‘Unpredictability and loss of control’, ‘Anxiety about the future’, ‘Body image and stigma’, ‘Physical stress’, ‘Optimism’, ‘Social situations’, ‘Family and partner’, ‘Work’ and ‘Healthcare’. Conclusion: People with IBD experience both physical and psychological stress, which requires a holistic view in nursing. Individualized care is crucial to maintaining trust and promoting positive outcomes for a person's treatment and health. Nurses should be attentive to individual needs and consider the psychological impacts of the disease. Additionally, creating a safe environment is important, as nurses can act as a supportive factor in case of social problems. The literature review highlights the significant and valuable role of nursing care in the experiences of individuals living with IBD.

Introduction Inflammatory bowel disease (IBD) and its complications have been well-established. The literature shows an association between IBD and decreased bone mineral density in the adult population. However, most studies have reported an association between IBD and osteoporosis, while the risk of fractures has not been well-studied. The aim of this meta-analysis is to summarize the best available evidence regarding IBS and osteoporotic fractures. Methods A review of the literature using the MEDLINE and EMBASE databases was performed during November 2017. We included cross-sectional and cohort studies that reported the relative risks, odds ratios, and hazard ratios comparing the risk of developing osteoporotic fractures among patients with IBD patients, both ulcerative colitis (UC) and Crohn's disease (CD), versus patients without IBD as controls. The pooled odds ratio (OR) and 95% confidence interval (CI) were calculated using the generic inverse-variance method. Results After a review of the literature, seven studies fulfilled the eligibility criteria established during the analysis. A significant association was found between IBD and osteoporosis, with a pooled OR of 1.32 (95% CI, 1.2 - 1.4). Low heterogeneity among the studies was found, I=42.3. No publication bias was found using the Egger regression test p=0.18. Sensitivity analysis showed that the inclusion of data on children by Kappelman et al. (2007) did not change the results. Conclusion A significant association between IBD and the risk of developing osteoporotic fractures was observed in this study. There is a 32% increased risk, which is consistent with different cohort studies previously done.

Colitis ulcerosa (CU) und Morbus Crohn (MC) zählen zur Gruppe der chronischen Darmerkrankungen (CED). Im Gegensatz zu CU lässt sich bei MC eine transmurale Entzündung und eine Ummantelung des entzündeten Dünndarms mit mesenterialen Fettgewebe, dem sogenannten “creeping fat”, feststellen. In dieser Arbeit wurden zwei verschiedene Mechanismen der Immunregulation in der Pathogenese von CED untersucht: 1) Der SOCE-Signalweg (store-operated Ca2+ entry) stellt eine wichtige Signalkaskade dar, die die Aktivierung von T-Zellen steuert. Wir haben die Auswirkung einer pharmakologischen Blockade von SOCE auf die Funktion von Immunzellen untersucht, die aus der intestinalen Mukosa therapierefraktärer CED-Patienten isoliert wurden. Anschließend konnten wir die Sicherheit einer systemischen Verabreichung von SOCE-Inhibitoren in vivo im einem murinen IBD-Modell bestätigen und zeigen, dass die Blockade von SOCE eine therapeutische Option für die Behandlung von CED darstellen könnte. 2) Darüber hinaus untersuchten wir, ob das Fehlen von Fettgewebe eine entzündungsfördernde oder -hemmende Rolle bei der Entstehung von CED spielt. Daher wurde die Zusammensetzung des mukosalen Immunsystems sowie die Funktion der intestinalen Epithelbarriere in einem Mausmodell mit Adipozytenatrophie sowohl im steady-state als auch nach induzierter Kolitis verglichen. Wir konnten zeigen, dass eine Fettgewebsatrophie vor dem Ausbruch einer Kolitis schützt, und führte zu einer erhöhten Resistenz der intestinalen Barriere. Schließlich verglichen wir die Merkmale des lipoatrophischen Mausmodells mit denen eines seltenen Patienten mit erworbener Lipodystrophie und MC und kamen zu dem Schluss, dass die chirurgische Resektion von mesenterialen Fettgewebes für Patienten mit einem MC, bei denen eine intestinale Resektionen durchgeführt wird, sinnvoll sein könnte. / Inflammatory bowel disease (IBD) is a group of chronic intestinal autoimmune disorders, including ulcerative colitis (UC) and Crohn's disease (CD). In contrast to UC, CD is characterized by transmural inflammation and mesenteric adipose tissue wrapping the inflamed small intestine, known as "creeping fat." Despite all currently available drug therapies, treating IBD remains a major challenge, underlying the necessity of identifying new therapeutic targets. In this work, two different mechanisms of immune regulation in the pathogenesis of IBD were investigated: First, because the store-operated Ca2+ entry (SOCE) signaling pathway is a crucial Ca2+ signaling cascade for T cell activation, we investigated the effect of pharmacological SOCE-blockade on intestinal immune cells isolated from therapy refractory IBD patients. Subsequently, we confirmed the efficacy and safety of systemic administration of SOCE inhibitors in vivo in an IBD murine model, demonstrating that the blockade of SOCE may represent a therapeutic option for treating IBD. Second, we investigated whether adipose tissue plays a pro- or anti-inflammatory role in the development of IBD. Therefore, we characterized the mucosal immune system and intestinal epithelial barrier in a murine model affected by adipocyte atrophy both at steady-state and after induction of colitis. We demonstrated that lipodystrophy protected against the onset of colitis and increased intestinal barrier resistance. Finally, we compared the lipoatrophic mouse model with a rare patient with acquired generalized lipodystrophy and CD, concluding that adipokines might play a pro-inflammatory role in IBD. Therefore, we suggest that surgical resection of mesenteric adipose tissue in CD patients might be a beneficial intervention in patients undergoing bowel resection.

Environmental factors such as diet contribute to the pathogenesis of inflammatory bowel disease (IBD). Epidemiological evidence suggests a robust linkage between IBD and the Western diet, which is often characterized by a high intake of food additives. These additives, including synthetic colourants, are widely used, leading to significant human exposure. Allura Red AC (AR) is one of the most popular synthetic colourants, yet little is known about its impact on human health and the role of AR in the pathogenesis of colitis remains elusive. Serotonin (5-hydroxytryptamine; 5-HT), which regulates various gut physiological processes, has been shown to modulate the gut microbiota and enhance susceptibility to colitis. In this thesis, it was discovered that chronic exposure to AR, at a dose found in commonly consumed dietary products, exacerbated dextran sulfate sodium (DSS)-induced colitis and triggered early onset of disease in the CD4+CD45RBhigh T cell-induced colitis model. AR also induced low grade colonic inflammation in naïve C57BL/6 mice. Exposure to AR was associated with increased colonic 5-HT levels and impaired intestinal barrier function via activation of the myosin light chain kinase (MLCK) pathway. However, AR did not promote colitis in mice lacking tryptophan hydroxylase 1 (Tph1), the rate-limiting enzyme responsible for colonic 5-HT synthesis. Further, AR increased colonic 5-HT levels in germ-free (GF) mice and perturbed the gut microbiota composition in specific pathogen-free (SPF) mice. Transfer of this altered microbiota from the dye-exposed SPF mice to GF mice conferred enhanced susceptibility to DSS-induced colitis. Mechanistically, AR induced reactive oxygen species (ROS) generation and promoted 5-HT secretion via the NF-κB pathway in BON cells. Data in this thesis indicate that the widely used synthetic colourant, AR, promotes colitis via colonic 5-HT in microbiota-dependent and -independent pathways. Collectively, these findings provide important information on enhancing public awareness of its detrimental effects on human health. / Thesis / Candidate in Philosophy / Epidemiological and experimental studies suggest a potential link between inflammatory bowel disease (IBD) and diet. The Western diet, often characterized by a high intake of processed foods, is associated with the growing incidence of IBD. Allura Red AC (AR) is a popular artificial food dye found in highly common processed foods, yet little is known about its impact on human health and disease. Serotonin, a key molecule in the gut, has been implicated in large bowel inflammation. Herein, the potential role of AR in the development of colitis was examined. Across multiple models, AR exposure heightened vulnerability to colitis in mice, an effect attenuated by reduced serotonin production in the gut. The effect of AR in enhancing colitis vulnerability occurred via gut microbiota-dependent and -independent pathways. These studies have identified how AR promotes colitis, findings that may advance public health awareness and impact the health of patients with IBD.