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Using blogs to describe Intensive Care nurses’ experiences of caring for patients in the COVID-19 pandemic. Background: In 2019, coronavirus disease 2019 (COVID-19) broke out worldwide, causing a pandemic. Research shows that patients with COVID-19 in intensive care units (ICUs) require more nursing care than other patients in ICU. The ICU nurses who care for these patients have shown signs of psychological and physical strain and experienced a lower sense of work satisfaction and feelings of helplessness. Analysis of blog posts has been conducted, to illuminate ICU nurses’ experiences of caring for patients with COVID-19. Method: To acquire a deeper understanding of ICU nurses’ experiences of caring for patients with COVID-19 during the first wave of the pandemic, analyzing text from ICU nurses’ blog posts was suitable. Seventy blog posts from 13 bloggers in the United States, Great Britain, Finland, and Sweden were analyzed using a manifest qualitative content analysis. Results: The results revealed an overall theme: “An overturned existence under extreme conditions.” Furthermore, three categories – “The virus caused changes in work and private life,” “Unreasonable demands,” and “To hold on to caring ideals thanks to the support of others”, were identified. Conclusion: Collecting data through blogs makes it possible to obtain information from a wide geographic area at almost no cost. Based on the overwhelming situation at ICUs, during the pandemic, analyzing blogposts was suitable to catch ICU nurses experiences without taking any time or focus from the participants during their work. Caring for patients with COVID-19 during the first wave of the pandemic was demanding, and support from colleagues and teamwork were important.

Background: The COVID-19 pandemic continues to impact health care workers’ (HCWs’) mental health. Research show that psychological distress and hard challenges abide with strong commitment to contribute. Yet, in-depth understanding of HCWs’ experiences of the changed caring reality during the pandemic is missing. Mixed methods research (MMR) accommodates several features apart from employing either quantitative or qualitative methods. This presentation focuses on examples from the 13-step mixed method by Leech and Onwuegbuzie. Methods: 97 HCWs in one hard-hit region in Sweden answered a web-based questionnaire measuring symptoms of stress (using DSM-5 criteria for post-traumatic stress syndrome, PTSD) in relation to perceived sense of coherence (SOC-scale) and self-compassion (SCS) as well as HCWs’ experiences. First, qualitative data (experiences of the pandemic) was analyzed by qualitative content analysis, then quantitative data (associations between PTSD and SOC/SCS) were analyzed with linear regression adjusted for covariates. Thirdly, a synthesis, the meta-inference of qualitative and quantitative data, explained a new comprehensive understanding. Results: By analyzing the categories and subcategories from the qualitative analysis in relation to symtoms of stress and SOC/SCS, a synthesis emerged. This was undertaken by merging and comparing the findings and discussing the new comprehensive understanding. Finally, to fully outline the mixed methods approach, qualitative and quantitative data were synthesized into a new comprehensive whole, a meta inference. Conclusion: This study moves away from dichotomic traditions between qualitative or quantitative approaches. By broadening the methodological departure this study may provide a new comprehensive understanding and contribute to enhance quality in MMR., nurses’ lived experiences focusing on the meaning of stress and caring in hospital emergency settings during the COVID-19 pandemic.

Background In 2019, coronavirus disease 2019 (COVID-19) broke out worldwide, leading to a pandemic. Studies have shown that COVID-19 patients in intensive care units (ICUs) require more nursing care than other patients. ICU nurses who care for patients with COVID-19 have shown signs of psychological and physical strain.AimThe aim of this study was to illuminate ICU nurses' experiences of caring for patients with COVID-19 in ICUs during the first wave of the pandemic.DesignA qualitative, descriptive and inductive approach was used. Method A total of 70 blog posts from 13 bloggers in the United States, Great Britain, Finland and Sweden were analysed using qualitative inductive manifest content analysis. Results The results reveal an overall theme: ‘An overturned existence under extreme conditions’. Furthermore, three categories—‘the virus caused changes in work and private lives’, ‘unreasonable demands’, and to hold on to caring ideals thanks to the support of others’—and seven subcategories were identified. Conclusion Caring for patients with COVID-19 during the first wave of the pandemic was demanding because of a lack of knowledge about the disease and the severity of the illness. This led to ICU nurses experiencing extreme conditions that affected various aspects of their lives. Support from colleagues and teamwork were revealed to be particularly important for how nurses dealt with the demands of working during a pandemic, as was sufficient recovery time between work shifts. Relevance to Clinical Practice Work in ICUs was challenging and demanding, even before the pandemic. This study contributes to an understanding of the complex work environment that existed in hospitals during the first wave of the COVID-19 pandemic. The knowledge obtained from this study can be used to revise working conditions and identify health interventions for ICU nurses.

Aim: To illuminate the meaning of newly graduated registered nurses' experiences of caring for patients in emergency departments during the COVID-19 pandemic. Design: A phenomenological hermeneutical study guided by Lindseth and Norberg. Methods: In-depth one-on-one interviews with 14 nurses from five hospitals were conducted from March to November 2020 and analysed using thematic analysis. The consolidated criteria for reporting qualitative research (COREQ) were used as the reporting guideline. Results: The findings comprise one main theme Caring through barriers and three themes with sub-themes. In the first theme, having intention to care, participants revealed their dedication to care for patients during the pandemic despite extensive stress, little experience and skills. The second theme, with tied hands in human suffering, illuminates experiences of being disconnected from the patient, overwhelmed by responsibility and unable to relieve suffering. The third theme, feeling inadequate, reveals experiences of lack of support and doubts meaning less space to develop into the nurse one wants to be. Conclusion: Findings reveal a new understanding of new nurses' experiences during times of crisis. The essence of caring in the emergency department during the pandemic can be explained as mediated through spatial, temporal and emotional barriers preventing new nurses from providing holistic care. Impact: The results may be used as anticipatory guidance for new nurses and inform targeted support interventions to support new nurses entering the profession in crisis conditions. Public Contribution: This study involved new nurses in semi-structured interviews.

Aim: To describe newly graduated registered nurses' (NGRNs') experiences of encountering stress in emergency departments (EDs) during the COVID-19 pandemic. Design: A qualitative descriptive study. Methods: Data from 14 in-depth interviews with NGRNs working in an ED for 3-36 months after graduation was analysed by the means of qualitative content analysis as described by Graneheim and Lundman. Interviews were conducted from March to November 2020 covering the first two waves of the pandemic. Results: Data revealed three categories and nine subcategories comprised in the theme Battling extraordinary situations and conflicting emotions. Empowered by acknowledging themselves as important caregivers during the pandemic NGRNs struggle against limitations and exert themselves beyond their known limit. External stressors due to work overload in combination with understaffing force NGRNs into the role of the experienced nurse prematurely and internal stressors derives from part taking in less qualitative care.

Aim Previous research has mainly focused on the advantages of PCC and less on its disadvantages. Hence, there is a need to further explore the recent research regarding PCC from both sides. Therefore, the aim of this paper is to elucidate the advantages and disadvantages of PCC. Design Discussion paper. Methods We searched relevant literature published January 2000-March 2018 in PubMed, Medline, CHINAL, Scopus and Web of Science. Results The results showed that PCC can contribute toimproved health and well-being, improved mutual interaction in relationships, improved cost-effectiveness and improved work environment, while the disadvantages can involveincreased personal and financial costs, exclusion of certain groups, increased personal and financial costs, exclusion of staff's personhood and unfairness due to empathy. An analysis of the existing literature on PCC showed paradoxes, which call for further investigation.

Telephone nurses give advice and support and make assessments based on verbal communication only. Web-based decision support systems are often used to increase patient safety and make medically correct assessments. The aim of the present this study was to describe factors affecting the use of a decision support system and experiences with this system among telephone nurses in Swedish primary health care. Observations and semistructured interviews were conducted. Six registered nurses with at least 1 year of experience of telephone nursing participated. Field notes and interviews were analyzed by qualitative content analysis. The main findings of the present this study were factors that decrease the decision support system use or promote deviation from decision support system use, factors that are positive for decision support system use and the decision support system complicates the work. Underuse and deviations from decision support systems can be a safety risk, because decisions are based on too little information. Further research with observations of telephone nurses' use of decision support systems is needed to develop both telephone nursing and decision support systems.

Objective: To provide a synthesis of already synthesized literature on person-centered care and patient-centered care in order to identify similarities and differences between the two concepts. Methods: A synthesis of reviews was conducted to locate synthesized literature published between January 2000 and March 2017. A total of 21 articles deemed relevant to this overview were synthesized using a thematic analysis. Results: The analysis resulted in nine themes present in person-centered as well as in patient-centered care: (1) empathy, (2), respect (3), engagement, (4), relationship, (5) communication, (6) shared decision-making, (7) holistic focus, (8), individualized focus, and (9) coordinated care. The analysis also revealed that the goal of person-centered care is a meaningful life while the goal of patient-centered care is a functional life. Conclusions: While there are a number of similarities between the two concepts, the goals for person-centered and patient-centered care differ. The similarities are at the surface and there are important differences when the concepts are regarded in light of their different goals. Practice implications: Clarification of the concepts may assist practitioners to develop the relevant aspects of care. Person-centered care broadens and extends the perspective of patient-centered care by considering the whole life of the patient.

Results: Systemized nomenclature of medicine clinical terms (SNOMED CT) was chosen, and the annotations were linked to its codes and terms. Two hundred WSI were collected and annotated, resulting in 17,497 annotations, covering a total area of 302.19 cm2, equivalent to 107,7 gigapixels. Ninety-five unique SNOMED CT codes were used. The time taken to annotate a WSI varied fr

Thiopurines are widely used in the treatment of leukemia and inflammatory bowel diseases. Thiopurine metabolism varies among individuals because of differences in the polymorphic enzyme thiopurine methyltransferase (TPMT, EC 2.1.1.67), and to avoid severe adverse reactions caused by incorrect dosing it is recommended that the patients TPMT status be determined before the start of thiopurine treatment. This study describes the concordance between genotyping for common TPMT alleles and phenotyping in a Swedish cohort of 12,663 patients sampled before or during thiopurine treatment. The concordance between TPMT genotype and enzyme activity was 94.5%. Compared to the genotype, the first measurement of TPMT enzyme activity was lower than expected for 4.6% of the patients. Sequencing of all coding regions of the TPMT gene in genotype/phenotype discrepant individuals led to the identification of rare and novel TPMT alleles. Fifteen individuals (0.1%) with rare or novel genotypes were identified, and three TPMT alleles (TPMT*42, *43, and *44) are characterized here for the first time. These 15 patients would not have been detected as carrying a deviating TPMT genotype if only genotyping of the most common TPMT variants had been performed. This study highlights the benefit of combining TPMT genotype and phenotype determination in routine testing. More accurate dose recommendations can be made, which might decrease the number of adverse reactions and treatment failures during thiopurine treatment., Funding Agencies|Swedish Childrens Cancer Foundation; Medical Research Council of Southeast Sweden; Swedish Society of Medicine Linkoping; ostgOtaregionens cancerfond

The dataset consists of 101 H&E-stained colon whole slide images (WSI) - 52 abnormal and 49 benign cases. All significant abnormal findings identified are outlined and categorized into 15 types such as hyperplastic polyp, high grade adenocarcinoma and necrosis. Other tissue components such as mucosa, submucosa, as well as the surgical margin are delineated to create a complete histological map. In total, 756 separate annotations have been made to segment the different tissue structures and link them to ontological information., One physician was responsible for the manual annotations controlled by a second pathologist. Accurate annotations were made over the whole tissues. 756 separate annotations were made.Following abnormal findings were annotated for the malign cases: acute and chronic inflammation, acute inflammation, adenocarcinoma, atrophy, chronic inflammation, diverticula, diverticulitis, dysplasia, edema, fibrosis, granulations tissue, hemorrhage, hyalinization, hyperplasia, hyperplastic polyp, inflammation, lymphoma, mucinous adenocarcinoma, necrosis, serrated adenoma, stasis, tubular adenoma, tubulovillous adenoma and ulceration.Other areas annotated: abnormal, artifact, cecum, colon, colonic mucous membrane, colonic muscularis propria, colonic submucosa, colonic subserosa, descending colon, ileum, normal, rectum, sigmoid colon and transverse colon.For the benign cases following areas were annotated: artifact, cecum, colon, colonic mucous membrane, colonic muscularis propria, colonic submucosa, colonic subserosa, descending colon, ileum, normal, rectum, sigmoid colon and transverse colon.

Background Undergraduate nurse education needs to prepare student nurses to meet the demands and to have the necessary communication skills for caring for an increasing older population. The challenges involve how best to support and empower student nurses to learn the communication skills needed to care for older people. Objective The aim of this study was to investigate student nurses' views on the care of and communication with older people. Design A descriptive study with a mixed-method approach was conducted. Methods Quantitative and qualitative data were collected from a questionnaire completed by third-year Swedish student nurses in 2015. Results The student nurses reported positive attitudes to the care of and communication with older people. The findings focus on the central aspects related to relationship building, techniques for communication and external prerequisites. Conclusions Despite positive attitudes, student nurses had a limited view of communication with older people. Educators need to increase student nurses' capacity to communicate effectively with older people. Educational interventions to improve and evaluate the communication competency of nurses and student nurses are needed.

Oligodendrocyte (OL) death may contribute to white matter pathology, a common cause of network dysfunction and persistent cognitive problems in patients with traumatic brain injury (TBI). Oligodendrocyte progenitor cells (OPCs) persist throughout the adult CNS and may replace dead OLs. OL death and OPCs were analyzed by immunohistochemistry of human brain tissue samples, surgically removed due to life-threatening contusions and/or focal brain swelling at 60.6 ± 75 hours (range 4–192 hours) postinjury in 10 severe TBI patients (age 51.7 ± 18.5 years). Control brain tissue was obtained postmortem from 5 age-matched patients without CNS disorders. TUNEL and CC1 co-labeling was used to analyze apoptotic OLs, which were increased in injured brain tissue (p < 0.05), without correlation with time from injury until surgery. The OPC markers Olig2, A2B5, NG2, and PDGFR-α were used. In contrast to the number of single-labeled Olig2, A2B5, NG2, and PDGFR-α-positive cells, numbers of Olig2 and A2B5 co-labeled cells were increased in TBI samples (p < 0.05); this was inversely correlated with time from injury to surgery (r = -0.8, p < 0.05). These results indicate that severe focal human TBI results in OL death and increases in OPCs postinjury, which may influence white matter function following TBI.

BACKGROUND: CYP3A metabolic activity varies between individuals and is therefore a possible candidate of influence on the therapeutic outcome of the tyrosine kinase inhibitor imatinib in chronic myeloid leukemia (CML) patients. The aim of this study was to investigate the influence of CYP3A metabolic activity on the plasma concentration and outcome of imatinib in CML patients. METHODS: Forty-three CML patients were phenotyped for CYP3A activity using quinine as a probe drug and evaluated for clinical response parameters. Plasma concentrations of imatinib and its main metabolite, CGP74588, were determined using liquid chromatography-mass spectrometry. RESULTS: Patients with optimal response to imatinib after 12 months of therapy did not differ in CYP3A activity compared to non-optimal responders (quinine metabolic ratio of 14.69 and 14.70, respectively; P=0.966). Neither the imatinib plasma concentration nor the CGP74588/imatinib ratio was significantly associated with CYP3A activity. CONCLUSIONS: CYP3A activity does not influence imatinib plasma concentrations or the therapeutic outcome. These results indicate that even though imatinib is metabolized by CYP3A enzymes, this activity is not the rate-limiting step in imatinib metabolism and excretion. Future studies should focus on other pharmacokinetic processes so as to identify the major contributor to patient variability in imatinib plasma concentrations.

Noninvasive tracking of biomaterials is vital for determining the fate and degradation of an implant in vivo, and to show its role in tissue regeneration. Current biomaterials have no inherent capacity to enable tracing but require labeling with, for example, fluorescent dyes, or nanoparticles. Here a novel biocompatible fully conjugated electrospun scaffold is described, based on a semiconducting luminescent polymer that can be visualized in situ after implantation using fluorescence imaging. The polymer, poly [2,3-bis-(3-octyloxyphenyl)quinoxaline-5,8-diyl-alt -thiophene-2,5-diyl] (TQ1), is electrospun to form a fibrous mat. The fibers display fluorescence emission in the near-infrared region with lifetimes in the sub-nanosecond range, optimal for in situ imaging. The material shows no cytotoxic behaviors for embryonic chicken cardiomyocytes and mouse myoblasts, and cells migrate onto the TQ1 fibers even in the presence of a collagen substrate. Subcutaneous implantations of the material in rats show incorporation of the TQ1 fibers within the tissue, with limited inflammation and a preponderance of small capillaries around the fibers. The fluorescent properties of the TQ1 fibers are fully retained for up to 90 d following implantation and they can be clearly visualized in tissue using fluorescence and lifetime imaging, thus making it both a pro-angiogenic and traceable biomaterial., Funding Agencies|Linkoping University; Swedish Foundation for Strategic Research; Swedish Research Council

ObjectiveThe tyrosine kinase inhibitors (TKIs) used in the treatment of chronic myeloid leukemia are substrates for the efflux transport protein ATP-binding cassette subfamily G member 2 (ABCG2). Variations in ABCG2 activity might influence pharmacokinetics and therapeutic outcome of TKIs. The role of ABCG2 single-nucleotide polymorphisms (SNPs) in TKI treatment is not clear and functional in-vitro studies are lacking. The aim of this study was to investigate the consequences of ABCG2 SNPs for transport and efficacy of TKIs [imatinib, N-desmethyl imatinib (CGP74588), dasatinib, nilotinib, and bosutinib].Materials and methodsABCG2 SNPs 34Ggreater thanA, 421Cgreater thanA, 623Tgreater thanC, 886Ggreater thanC, 1574Tgreater thanG, and 1582Ggreater thanA were constructed from ABCG2 wild-type cDNA and transduced to K562 cells by retroviral gene transfer. Variant ABCG2 expression in cell membranes was evaluated and the effects of ABCG2 SNPs on transport and efficacy of TKIs were measured as the ability of ABCG2 variants to protect against TKI cytotoxicity.ResultsWild-type ABCG2 had a protective effect against the cytotoxicity of all investigated compounds except bosutinib. It was found that ABCG2 expression provided better protection against CGP74588 than its parent compound, imatinib. ABCG2 421Cgreater thanA, 623Tgreater thanC, 886Ggreater thanC, and 1574Tgreater thanG reduced cell membrane expression of ABCG2 and the protective effect of ABCG2 against imatinib, CGP74588, dasatinib, and nilotinib cytotoxicity.ConclusionThese findings show that the ABCG2 SNPs 421Cgreater thanA, 623Tgreater thanC, 886Ggreater thanC, and 1574Tgreater thanG increase the efficacy of investigated TKIs, indicating a reduced transport function that might influence TKI pharmacokinetics in vivo. Furthermore, the active imatinib metabolite CGP74588 is influenced by ABCG2 expression to a greater extent than the parent compound.

Single-nucleotide polymorphisms (SNPs) in the gene coding for the efflux-transport protein ABCB1 (P-glycoprotein) are commonly inherited as haplotypes. ABCB1 SNPs and haplotypes have been suggested to influence the pharmacokinetics and therapeutic outcome of the tyrosine kinase inhibitor (TKI) imatinib, used for treatment of chronic myeloid leukemia (CML). However, no consensus has yet been reached with respect to the significance of variant ABCB1 in CML treatment. Functional studies of variant ABCB1 transport of imatinib as well as other TKIs might aid the interpretation of results from in vivo association studies, but are currently lacking. The aim of this study was to investigate the consequences of ABCB1 variant haplotypes for transport and efficacy of TKIs (imatinib, its major metabolite N-desmethyl imatinib [CGP74588], dasatinib, nilotinib, and bosutinib) in CML cells. Variant haplotypes - including the 61A>G, 1199G>A, 1236C>T, 1795G>A, 2677G>T/A, and 3435T>C SNPs - were constructed in ABCB1 complementary DNA and transduced to K562 cells using retroviral gene transfer. The ability of variant cells to express ABCB1 protein and protect against TKI cytotoxicity was investigated. It was found that dasatinib and the imatinib metabolite CGP74588 are effectively transported by ABCB1, while imatinib, nilotinib, and bosutinib are comparatively weaker ABCB1 substrates. None of the investigated haplotypes altered the protective effect of ABCB1 expression against TKI cytotoxicity. These findings imply that the ABCB1 haplotypes investigated here are not likely to influence TKI pharmacokinetics or therapeutic efficacy in vivo.

Karin Skoglund,1 Samuel Boiso Moreno,1 Maria Baytar,1 Jan-Ingvar Jönsson,2 Henrik Gréen1,3 1Department of Medical and Health Sciences, 2Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden; 3Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden Abstract: Single-nucleotide polymorphisms (SNPs) in the gene coding for the efflux-transport protein ABCB1 (P-glycoprotein) are commonly inherited as haplotypes. ABCB1 SNPs and haplotypes have been suggested to influence the pharmacokinetics and therapeutic outcome of the tyrosine kinase inhibitor (TKI) imatinib, used for treatment of chronic myeloid leukemia (CML). However, no consensus has yet been reached with respect to the significance of variant ABCB1 in CML treatment. Functional studies of variant ABCB1 transport of imatinib as well as other TKIs might aid the interpretation of results from in vivo association studies, but are currently lacking. The aim of this study was to investigate the consequences of ABCB1 variant haplotypes for transport and efficacy of TKIs (imatinib, its major metabolite N-desmethyl imatinib [CGP74588], dasatinib, nilotinib, and bosutinib) in CML cells. Variant haplotypes – including the 61A>G, 1199G>A, 1236C>T, 1795G>A, 2677G>T/A, and 3435T>C SNPs – were constructed in ABCB1 complementary DNA and transduced to K562 cells using retroviral gene transfer. The ability of variant cells to express ABCB1 protein and protect against TKI cytotoxicity was investigated. It was found that dasatinib and the imatinib metabolite CGP74588 are effectively transported by ABCB1, while imatinib, nilotinib, and bosutinib are comparatively weaker ABCB1 substrates. None of the investigated haplotypes altered the protective effect of ABCB1 expression against TKI cytotoxicity. These findings imply that the ABCB1 haplo

Objectives: The "neurological wake-up test" is needed to evaluate the level of consciousness in patients with severe traumatic brain injury. However, the neurological wake-up test requires interruption of continuous sedation and may induce a stress response and its use in neurocritical care is controversial. We hypothesized that the neurological wake-up test induces an additional biochemical stress response in patients with severe traumatic brain injury. Patients: Twenty-four patients who received continuous propofol sedation and mechanical ventilation after moderate to severe traumatic brain injury (Glasgow Coma Scale score <= 8; patient age 18-71 yrs old) were analyzed. Exclusion criteria were age <18 yrs old, ongoing pentobarbital infusion, or markedly increased intracranial pressure on interruption of continuous sedation. Design: Single-center prospective study. During postinjury days 1-8, 65 neurological wake-up tests were evaluated. Adrenocorticotrophic hormone, epinephrine, and norepinephrine levels in plasma and cortisol levels in saliva were analyzed at baseline (during continuous intravenous propofol sedation) and during neurological wake-up test. Data are presented using medians and 25th and 75th percentiles. Setting: The study was performed in a university hospital neurocritical care unit. Interventions: None. Measurements and Main Results: At baseline, adrenocorticotrophic hormone and cortisol levels were 10.6 (6.0-19.4) ng/L and 16.0 (10.7-31.8) nmol/L, respectively. Immediately after the neurological wake-up test, adrenocorticotrophic hormone levels increased to 20.5 (11.1-48.4) ng/L (p < .05) and cortisol levels in saliva increased to 24.0 (12.3-42.5) nmol/L (p < .05). The plasma epinephrine and norepinephrine levels increased from a baseline of 0.3 (0.3-0.6) and 1.6 (0.9-2.3) nmol/L, respectively, to 0.75 (0.3-1.4) and 2.8 (1.28-3.58) nmol/L, respectively (both p < .05). Conclusions: The neurological wake-up test induces a biochemical

The enzyme thiopurine S-methyltransferase (TPMT) is involved in the metabolism of thiopurine drugs used to treat acute lymphoblastic leukemia and inflammatory bowel disease. Thus far, at least 29 variants of the TPMT gene have been described, many of which encode proteins that have low enzyme activity and in some cases become more prone to aggregation and degradation. Here, the two naturally occurring variants, TPMT*2 (Ala80 → Pro) and TPMT*5 (Leu49 → Ser), were cloned and expressed in Escherichia coli. Far-UV circular dichroism spectroscopy showed that TPMT*2 was substantially destabilized whereas TPMT*5 showed much greater stability comparable to that of wild-type TPMT (TPMTwt). The extrinsic fluorescent molecule anilinonaphthalene sulfonate (ANS) was used to probe the tertiary structure during thermal denaturation. In contrast to TPMTwt, neither of the variants bound ANS to a large extent. To explore the morphology of the TPMT aggregates, we performed luminescent conjugated oligothiophene staining and showed fibril formation for TPMT*2 and TPMT*5. The differences in the flexibility of TPMTwt, TPMT*2, and TPMT*5 were evaluated in a limited proteolysis experiment to pinpoint stable regions. Even though there is only one amino acid difference between the analyzed TPMT variants, a clear disparity in the cleavage patterns was observed. TPMT*2 displays a protected region in the C-terminus, which differs from TPMTwt, whereas the protected regions in TPMT*5 are located mainly in the N-terminus close to the active site. In conclusion, this in vitro study, conducted to probe structural changes during unfolding of TPMT*2 and TPMT*5, demonstrates that the various causes of the low enzyme activity in vivo could be explained on a molecular level., funding agencies|Swedish Childhood Cancer Foundation||Swedish Research Council||EMBO