Pankreatit anses vara en vanligt förekommande sjukdom hos hundar och katter och kan diagnostiseras genom mätningar av koncentrationen Canine Pancreas-specific Lipase (cPL) respektive Feline Pancreas-specific Lipase (fPL) i serum. Utifrån dessa koncentrationer graderas patienten enligt normalvärde, gråzon eller indikation på pankreatit. Golden standardmetoden för att analysera cPL/fPL är Spec cPL respektive Spec fPL. Nya metoder har utvecklats för kvantitativ mätning av pankreas-specifik lipas såsom Vcheck V200, ett instrument, som analyserar cPL/fPL med en fluorescerande immunoassay. Syftet med studien var att utvärdera analys av cPL/fPL på Vcheck V200 samt jämföra resultatet från detta instrument med värdena från ett referenslaboratorium i Tyskland för att se om det fanns en signifikant skillnad mellan metoderna. Koncentrationen cPL i hundserum (n=37) och koncentrationen fPL i kattserum (n=29) analyserades på Vcheck V200. Dessa prover skickades även till referenslaboratoriet där analysen Spec cPL respektive Spec fPL utfördes. Spridningen var stor kring bias i Bland-Altman diagram för både cPL och fPL och jämförelsen mellan metoderna för de specifika koncentrationerna av cPL/fPL bedömdes vara statistiskt signifikant (p<0,05). 27% av hundproverna graderas olika enligt de båda metoderna och skillnaden var signifikant (p<0,05). 24% av kattproverna graderades olika men skillnaden var inte signifikant (p=0,257). Studien tyder på att jämförelsen mellan de båda metoderna var signifikant förutom vid graderingen av kattproverna. Beaktande detta och det faktum att kvalitetssäkringen brister vid analys av fPL på grund av avsaknad av kontroller kan i dagsläget inte cPL/fPL på Vcheck V200 ersätta den nuvarande golden standardmetoden, trots att ingen signifikant skillnad sågs vid gradering av kattprover. / Pancreatitis is a common disease in canine and felines and can be diagnosed by measuring the concentration of Canine Pancreas-specific Lipase (cPL) or Feline Pancreas-specific Lipase (fPL) in serum. Based on the concentration of cPL/fPL, the patient is then classified in different diagnostic categories (normal value, gray zone or indication of pancreatitis). Spec cPL and Spec fPL is currently the golden standard method for analysis of cPL and fPL. New methods have been developed for the quantitative measurement of pancreatic lipases. Vcheck V200, being one example, utilizing a fluorescent immunoassay for quantification of the lipase. The aim of this study was to evaluate the cPL and fPL analysis on the Vcheck V200 and to examine if there was a significant difference (p≤0,05) when comparing the result from Vcheck V200 with the results from a reference laboratory. The concentration of cPL (n=37) and fPL (n=29) in serum from canine or felines were analyzed using Vcheck V200. The samples were also sent to the reference laboratory where Spec cPL and Spec cPL were performed. A Bland-Altman plot comparison between the two methods showed a large spread for both analysis of cPL and fPL. Comparison of the specific values for analysis of cPL and fPL between the two methods revealed a significant difference (p<0,05). 27% of the dog samples were categorized differently according to the two methods and this difference was significant (p<0,05). 24% of the cat samples were categorized differently and no significant difference were observed (p=0,257). This study indicates that the difference between the two methods was significant, besides the classification of cat samples. Considering this and the lack of quality assurance regarding analysis of fPL due to lack of controls, the cPL/fPL analysis on Vcheck V200 cannot replace the Spec cPL or Spec fPL at present.

POPULAR SCIENTIFIC SUMMARY Systemic lupus erythematosus (SLE) is not your average disorder. It behaves like a mischievous troublemaker, wreaking havoc throughout the body, causing inflammation that affects multiple organs. SLE presents a puzzle that keeps health care professionals worldwide intrigued, searching for answers amidst its complex of immunologic manifestations and clinical symptoms. While we’ve made progress in understanding SLE, its specific cause remains a mystery. What we do know is that SLE triggers a fascinating interplay between genetic, hormonal, and environmental factors in susceptible individuals. Macrophages, specialized white blood cells, can be likened to moody actors on a stage wearing different masks and wielding functional props. Among them are M1 macrophages, fiery troublemakers who provoke pro-inflammatory responses, and M2 macrophages, peacemakers striving for balance by generating anti-inflammatory responses. Then there is NRF2, the vigilante, normally held by its captor, KEAP1. However, when cells stress NRF2 manages to break free from KEAP1 and spring into action, embarking on a crucial journey into the cell nucleus where DNA is stored. Once inside, NRF2 binds specific regions of the DNA, promoting genes associated with protective activities, including antioxidative responses and detoxification processes, thereby shielding cells from further harm. Now, let us envision a therapeutic strategy that utilizes this; if we can deliberately unleashNRF2 on command, triggering a powerful cascade of antioxidative responses throughout the body,such a treatment would offer tremendous promise and serve as a paradigm for patients sufferingfrom chronic inflammation. But the question remains: Is it possible? In this study, we investigated the effects of certain chemicals on macrophages in a controlledlab environment. Our goal was to explore their potential for therapeutic purposes. Excitingly, wediscovered that these chemicals can indeed influence macrophages to produce a stronger antiinflammatory and antioxidant response. These findings could be promising for developing futuretreatments, especially in patients diagnosed with conditions such as SLE. / ABSTRACT Systemic lupus erythematosus (SLE) is a multifaceted, chronic autoimmune disorder that leads to inflammation and affects various organs. A wide range of immunologic manifestations and clinical symptoms characterizes SLE. While the specific cause remains unknown, it is thought to result from a combination of genetic susceptibility and the intricate interplay between environmental and hormonal factors. A significant subset of SLE patients also experience renal manifestation, lupus nephritis (LN), characterized by distinct inflammatory responses in which macrophages play a role. Macrophages exhibit different functional characteristics depending on their environment, and generally display two contrasting phenotypes; M1, which elicits proinflammatory responses, and M2, which generates anti-inflammatory responses Homeostasis is vital, yet environmental stress is inevitable. NRF2, a transcription factor known for its involvement in oxidative stress response, plays a pivotal role. Under basal conditions, NRF2 resides in the cytoplasm and is targeted for degradation by the protein KEAP1. However, during cellular stress, the NRF2-KEAP1 complex dissociates, allowing NRF2 to translocate into the nucleus where it binds specific regulatory regions of genes that promote cytoprotective activities. The NRF2 pathway has gained attention as a potential target for therapeutic strategies in inflammatory conditions, including SLE. This study aimed to assess the effects of certain chemical NRF2 activators and a KEAP1 inhibitor on an in vitro model of M1 and M2 macrophage polarization. The objective was to investigate whether these compounds could enhance antioxidative response. To evaluate this, key genes and proteins involved in antioxidative pathways were analyzed. Gene expression was assessed using quantitative real-time PCR (qPCR), and protein presence was determined through immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA). The findings of this study indicate that stimulation of macrophage subgroups with the selected compounds promotes a shift towards anti-inflammatory and antioxidative response. / <p>Rektor tilldelade Leo Svahn stipendie Österby för <em>välartade obemedlade studier</em>.</p>

Chronic pain affects 20 % of the global population, causes suffering, is difficult to treat, and constitutes a large economic burden for society. So far, the characterization of molecular mechanisms of chronic pain-like behaviors in animal models has not translated into effective treatments. In this thesis, consisting of five studies, pain patient biofluids were analyzed with modern proteomic methods to identify biomarker candidates that can be used to improve our understanding of the pathophysiology chronic pain and lead to more effective treatments. Paper I is a proof of concept study, where a multiplex solid phase-proximity ligation assay (SP-PLA) was applied to cerebrospinal fluid (CSF) for the first time. CSF reference protein levels and four biomarker candidates for ALS were presented. The investigated proteins were not altered by spinal cord stimulation (SCS) treatment for neuropathic pain. In Paper II, patient CSF was explored by dimethyl and label-free mass spectrometric (MS) proteomic methods. Twelve proteins, known for their roles in neuroprotection, nociceptive signaling, immune regulation, and synaptic plasticity, were identified to be associated with SCS treatment of neuropathic pain. In Paper III, proximity extension assay (PEA) was used to analyze levels of 92 proteins in serum from patients one year after painful disc herniation. Patients with residual pain had significantly higher serum levels of 41 inflammatory proteins. In Paper IV, levels of 55 proteins were analyzed by a 100-plex antibody suspension bead array (ASBA) in CSF samples from two neuropathic pain patient cohorts, one cohort of fibromyalgia patients and two control cohorts. CSF protein profiles consisting of levels of apolipoprotein C1, ectonucleotide pyrophosphatase/phosphodiesterase family member 2, angiotensinogen, prostaglandin-H2 D-isomerase, neurexin-1, superoxide dismutases 1 and 3 were found to be associated with neuropathic pain and fibromyalgia. In Paper V, higher CSF levels of five chemokines and LAPTGF-beta-1were detected in two patient cohorts with neuropathic pain compared with healthy controls. In conclusion, we demonstrate that combining MS proteomic and multiplex antibody-based methods for analysis of patient biofluid samples is a viable approach for discovery of biomarker candidates for the pathophysiology and treatment of chronic pain. Several biomarker candidates possibly reflecting systemic inflammation, lipid metabolism, and neuroinflammation in different pain conditions were identified for further investigation. / Uppsala Berzelii Technology Centre for Neurodiagnostics