Genetic polymorphism is by definition the occurrence in the same population of two or more alleles at one locus,each with a frequency high enough not to be maintained by recurrent mutation only (1). Among the human plasmaproteins two major categories of polymorphism have been described, allotypic and electrophoretic heterogeneity. Allotypy is defined by Oudin as individual antigenic differences among proteins within a species (2). The first discovered polymorphism of this category was the Gm system of immunoglobulin G by Grubb (3). The first described electrophoretic heterogeneity in plasma proteins was theHp (haptoglobin) system discovered by Smithies (4). Sincethen genetic variants of several other human plasma proteins have been found. This dissertation is concerned with thegenetic and immunological aspects of the polymorphism of the third component of human complement, C3. / digitalisering@umu.se

In the last decades, the prevalence of obesity increased dramatically worldwide. According to the WHO, 2.1 billion people (30% of the population) around the world are obese or overweight. Effective ways to decrease food intake are needed. Evidence from emerging studies indicates an association between feeding behavior and the IL-6 expression in the central nervous system (CNS). The study aimed to investigate the effect of IL-6 modulation on feeding behavior and learning and memory process in the ventral hippocampus (vHPC). To implement this aim two groups of rats were used, the first group was exposed to the reduction of the IL-6 expression (knockdown), whereas the other one to microinjections of exogenous IL-6 (EX IL-6). Both experimental groups were subjected to a set of behavioral and molecular tests specific for investigating memory process, emotional/affective behavior and food intakes like novel object recognition, Morris water maze, and social interaction test. The results for IL-6 knockdown (KD) showed improvement in the short term memory, but did not affect the food intake. On the other hand, EX IL-6 caused an increase in the locomotor’s activity and the food intake during the 24 hours, but at the same time caused impairment in the spatial and learning memory. Taken together, these results provide new insight on the role of IL-6 outside of inflammation highlighting its ability to modulate hippocampus-dependent mnemonic process, and affective and feeding behaviors in the vHPC, however several questions still remain not addressed and the study require further investigation.

No description available.

Cerebral stroke has become one of the leading causes of death and disability worldwide. During an ischemic stroke, oxygen and nutrient deprivation occurs, which combined lead to cell starvation, anoxia, and eventually cell death. However, when blood flow is restored, reperfusion damage occurs resulting in increased cell death through several mechanisms. One of the main reasons behind ischemia/reperfusion damage is oxidative stress due to elevated production of reactive oxygen species (ROS) during reperfusion. There are several proteins and processes that are thought to be involved in elevated oxidative stress and the formation of ROS during reperfusion, among which the NADPH oxidase (Nox) family is suggested to be the main contributor of ROS.To examine this hypothesis, in the present work, we inhibited activity of the Nox2 and Nox4 enzymes during ischemia/reperfusion with the Glucox Biotech AB (Sweden) inhibitors M4, M107, and M114 to evaluate whether reducing Nox activity could reduce the ischemia/reperfusion-induced cell death, hence be used as a potential stroke treatment, the cell viability was measured with MTS after ischemia/reperfusion induction and treatment with the Nox substances. We also examined the gene expression levels of the Nox enzymes Nox2 and Nox4 with qPCR after induced ischemia/reperfusion in the neuroblastoma cell line NB69.Our results showed a decrease in Nox4 gene expression after 1h ischemia/8h reperfusion and an increased expression after 1h ischemia/24h reperfusion in NB69 cells. Treatment with M114 resulted in increased cell viability after 2h ischemia/72h reperfusion. However, the toxic effect of ischemia/reperfusion-induced response was found to be inadequate, as indicated by extensive proliferation and lack of cell death. This unfavorable outcome is suggested to be excess of oxygen in medium, metabolization of L-glutamine, and effects of growth factors in the serum used in cell culture medium during the ischemic phase. Therefore, the cell culture protocol was modified to the use of PBS instead of glucose-free medium under serum-free condition during the ischemia. The altered ischemic conditions resulted in continuous reduction in cell viability at increasing ischemic time points with total cell death at 2h ischemia, suggesting applicable conditions for ischemia/reperfusion studies. Even though a conclusion could not be made about the inhibitors M4, M107, and M114 as the cell viability assay was performed under insufficient conditions; the Nox inhibitors shows high potential as future ischemic stroke treatments, which may help save lives and improve life quality for affected patients.

No description available.

Nociceptive afferent axons of the stomach send signals centrally to the brain and locally to stomach tissues. Nociceptive axons can be detected with a variety of different markers. In particular, the neuropeptide substance P (SP) is one of the most commonly used markers for nociceptive nerves in the somatic and visceral organs. In addition, the Local release of SP contributes to tissue protection and repair in response to noxious stimuli. However, the topographical distribution and morphological structure of SP-immunoreactive (SP-IR) axons and terminals in the whole stomach have not yet been well determined. In this study, we labeled SP-IR axons and terminals in flat-mounts and serial sections of the antrum, pyloric sphincter, and duodenum (APD) of the mouse stomach. The flat-mount stomachs included the external muscles (longitudinal and circular layers) and the myenteric ganglionic plexus, whereas APD serial sections also included the submucosa and mucosa. Tissue samples were scanned using confocal and Zeiss M2 imager microscopy to acquire detailed high-resolution images of SP innervation and produce montages of the whole stomach flat-mount and APD serial sections. Additionally, we double-labeled the samples with vesicular acetylcholine transporter (VAChT) and tyrosine hydroxylase (TH), and injected the stomach walls with DiI-retrograde tracer to determine the connectivity map between SP-IR axons and their possible extrinsic origin. We found that: 1) SP-IR axons innervated all layers including the external muscles, myenteric ganglia, submucosa, submucosal ganglia, and mucosa. Many axons were immunoreactive for VAChT but not TH. 2) SP-IR axons and terminals formed an extensive network in the muscular layers and ran in parallel with the long axis of the external muscles. 3) SP-IR axons formed very dense terminal varicosities encircling individual neurons in the myenteric plexus 4) In the submucosa, SP-IR axons innervated blood vessels and submucosal ganglia and formed a nerve network around duodenal Brunner's glands. 5) In the mucosa, SP-IR varicose axons were organized into bundles in the muscularis mucosa and lamina propria at the base of the mucosa. Some SP-IR axons were found in the gastric subepithelial level and duodenal villi. 6) SP-IR axon density in the muscles and myenteric plexus was much higher than in the submucosa and mucosa. 7) The density of SP-IR axons and terminals in the flat-mounts varied in different stomach regions in the following order from high to low: antrum-pylorus, corpus, fundus, and cardia. 8) In the APD area, the muscular wall of the antrum and duodenum showed a higher SP-IR axon density than in the pyloric sphincter. 9) The extrinsic origin of SP-IR axons in the stomach derived mainly from the spinal T7-T11 dorsal root ganglia with a lesser contribution from the vagal nodose ganglia, but not from the celiac ganglia, the dorsal motor nucleus of vagus, or the nucleus tractus solitarius. Collectively, our data provide for the first time a map of the topographical distribution and morphology of SP-IR axons and terminals in the whole stomach with single-cell/axon/synapse resolution. This work will establish an anatomical foundation for functional mapping of the SP-IR nociceptive afferent innervation of the stomach and its pathological remodeling in gastrointestinal diseases.

Doxorubicin (Dox) has been one of the most effective chemotherapeutic drugs against a wide range of cancers. The use of Dox is restricted due to severe clinical side effects including Dox-induced skeletal muscle toxicity (DIMT). The exact molecular mechanisms in the progression and development of DIMT have not been established. We investigated the roles of inflammatory cell deaths pyroptosis and necroptosis in DIMT. Additionally, we explored the therapeutic potential of embryonic stem cell-derived exosomes (ES-Exos) in diminishing DIMT. Mice were used to develop DIMT and pre-diabetic animal models of muscle toxicity. Mice were subjected to muscle function and glucose tolerance testing. Soleus and gastrocnemius tissues were collected for protein expression assays including immunohistochemistry, western blot, ELISA, and RT-PCR to determine pyroptosis, inflammasome formation, M1 and M2 macrophages, necroptosis and necroinflammation levels. Histology staining was performed to determine atrophy and fibrosis. Muscle strength of mice were significantly (p < 0.05) diminished for Dox vs. controls whereas ES-Exos significantly (p < 0.05) mitigated this muscle dysfunction by enhancing muscle strength. In parallel, our results showed a significant (p < 0.05) increase in muscle atrophy, fibrosis, inflammasome formation leading to inflammation-induced cell death pyroptosis, necroinflammation, and pro-inflammatory M1 macrophage markers for Dox compared with controls. Treatment with ES-Exos significantly (p < 0.05) reduced atrophy, fibrosis, inflammation, inflammatory cytokines, and pyroptosis. Noticeably, ES-Exos treatment significantly enhanced anti-inflammatory M2 macrophages in the muscle. Necroptosis and necroinflammation markers, along with atrophy and fibrosis, were significantly (p < 0.05) increased in the combined model of Dox and pre-diabetes compared with pre-diabetes alone, suggesting that Dox administration in pre-diabetic conditions have enhanced detrimental effects. Overall, this novel study establishes the mechanistic roles of inflammatory cell deaths pyroptosis and necroptosis, inflammatory cytokines, and inflammatory macrophages in skeletal muscle toxicity. Moreover, we established the protective effects of Exos in muscle toxicity models involving Dox.

Lignocellulosic biomass has potential to chip in the chemical and biofuels supplies in future societies,even though lignocellulose is a recalcitrant structure that has to be treated in several steps. After theirproper life cycle, wood-derived materials such as particleboards have few outcomes today apart fromenergy recovery for heat production. Then, they may be used as lignocellulosic biomass sources in theproduction of molecules of interest. Fermentation from wood-derived monosaccharides imposespreliminary sugar retrieval, for instance through pre-treatment and enzymatic hydrolysis. This studyfocuses on the potential of particleboards waste for chemical and biofuel production by comparingsaccharification through simulated steam explosion pre-treatment and enzymatic hydrolysis betweennative and particleboard-derived wood, with an insight in subsequent fermentation by Saccharomycescerevisiae. Urea-Formaldehyde bound particleboard was investigated, as well as some aspects ofMelamine-Urea-Formaldehyde bound particleboard.Pre-treatment resulted in apparition of lignocellulosic degraded compounds in a much larger extent innative wood than in particleboard, which seemed to be only superficially impacted. Formation ofdegraded compounds from sugars – furfural and 5-hydroxymethylfurfural – was enhanced when pretreatmentwas prolonged. Removal of a substantial fraction of the adhesive contained in theparticleboards was observed, leading to comparable concentrations in free urea, its degradedproducts, and formaldehyde between native wood and particleboards during enzymatic hydrolysis.Enzymatic hydrolysis with cellulases and hemicellulases highlighted a critical role of pre-treatment toenhance final yields, both in native wood and in Urea-Formaldehyde particleboard. Adding 20 minutessteam-explosion type pre-treatment at 160 °C resulted in glucose yields increase from 18.5 % to 32.8% for native wood and from 15.6 % to 37.4 % for particleboard. Prolonging pre-treatment residencetime to 35 minutes resulted in much better glucose extraction for native wood but only slight progressfor the particleboard, as glucose yields reached 64.5 % and 41.1 % respectively. Maximalconcentrations achieved were 277 and 184 mg/gbiomass respectively.Fermentation brought to light high inhibition from both native wood and particleboard sources ofmedia, which were attributed to components or degraded products of lignocellulose that were notanalysed in this project. Ethanol was formed during fermentation, with reduced productivity butincreased yields as compared with the control sample. Inhibition was so strong that no difference couldbe given between native and particleboard wood. In this situation, no inhibition potential of resin orits degradation products could be proved. / Lignocellulosic biomassa har potential att bidra till kemikalier och biobränsletillförsel i framtidasamhällen, trots att lignocellulosa är en rekalcitrant struktur som måste behandlas i flera steg. Idagträmaterial som spånskivor bara används för energiåtervinning och värmeproduktion efter deraslivscykel. De kan därför användas som råvara för framställning av värdefulla molekyler.Fermenteringsprocesser behöver frisättningen av trä monosackarider genom förbehandlingsprocesseroch enzymatisk hydrolys. Studien fokuserar på potentialen för avfall från spånskivor för kemisk ochbiobränsleproduktion. Vi har jämfört sackarifiering mellan nativt trä och spånskivor genom simuleradångaxplosion och enzymatisk hydrolys, med en inblick i efterföljande fermentering av Saccharomycescerevisiae. Spånskivor bunden av urea-formaldehyd undersöktes, liksom vissa aspekter av spånskivorbundna med melamin-urea-formaldehyd.Förbehandlingen producerade högre koncentration av lignocellulosa nedbrytningsprodukter frånnativt trä jämfört med spånskivor. Bildningen av nedbrytningsprodukter från sockerarter - furfural och5-hydroxymethylfurfural - ökade med längre förbehandlingar. En väsentlig fraktion av limmet borttogsfrån spånskivorna, vilket ledde till jämförbara koncentrationer i fri urea, dess nedbrytningsprodukteroch formaldehyd mellan naturligt trä och spånskivor under enzymatisk hydrolys.Enzymatisk hydrolys med cellulaser och hemicellulaser avslöjade den kritiska rollen av förbehandlingför att förbättra utbytet, både i naturligt trä och i urea-formaldehyd spånskiva. Längre (20 minuter)ångexplosion vid 160° C resulterade i högre glukosutbytet (från 18,5% till 32,8% för naturligt trä ochfrån 15,6% till 37,4% för spånskivor). Förlängning av uppehållstiden före behandlingen till 35 minuterresulterade i mycket bättre glukosekstraktion för nativt trä (64,5%) men endast liten framsteg förspånskivan (41,1%). Detta resulterade i maximalt utbyte av 277 mg Glc/g biomassa och 184 mg Glc/ gbiomassa för nativt trä och spånskivor, respektive.Fermentering visade hög hämning från lignocellulosa nedbrytningsprodukter som inte analyserades iprojektet för både nativt trä och spånskällor för media. Etanol bildades under fermentering medreducerad produktivitet men ökade utbyten jämfört med kontrollprovet. Hämningen var så stark attingen skillnad kunde ges mellan naturligt trä och spånskivor. I denna situation kunde ingenhämningspotential för lim eller dess nedbrytningsprodukter bevisas.

Lignocellulosic biomass has potential to chip in the chemical and biofuels supplies in future societies,even though lignocellulose is a recalcitrant structure that has to be treated in several steps. After theirproper life cycle, wood-derived materials such as particleboards have few outcomes today apart fromenergy recovery for heat production. Then, they may be used as lignocellulosic biomass sources in theproduction of molecules of interest. Fermentation from wood-derived monosaccharides imposespreliminary sugar retrieval, for instance through pre-treatment and enzymatic hydrolysis. This studyfocuses on the potential of particleboards waste for chemical and biofuel production by comparingsaccharification through simulated steam explosion pre-treatment and enzymatic hydrolysis betweennative and particleboard-derived wood, with an insight in subsequent fermentation by Saccharomycescerevisiae. Urea-Formaldehyde bound particleboard was investigated, as well as some aspects ofMelamine-Urea-Formaldehyde bound particleboard. Pre-treatment resulted in apparition of lignocellulosic degraded compounds in a much larger extent innative wood than in particleboard, which seemed to be only superficially impacted. Formation ofdegraded compounds from sugars – furfural and 5-hydroxymethylfurfural – was enhanced when pretreatmentwas prolonged. Removal of a substantial fraction of the adhesive contained in theparticleboards was observed, leading to comparable concentrations in free urea, its degradedproducts, and formaldehyde between native wood and particleboards during enzymatic hydrolysis.Enzymatic hydrolysis with cellulases and hemicellulases highlighted a critical role of pre-treatment toenhance final yields, both in native wood and in Urea-Formaldehyde particleboard. Adding 20 minutessteam-explosion type pre-treatment at 160 °C resulted in glucose yields increase from 18.5 % to 32.8% for native wood and from 15.6 % to 37.4 % for particleboard. Prolonging pre-treatment residencetime to 35 minutes resulted in much better glucose extraction for native wood but only slight progressfor the particleboard, as glucose yields reached 64.5 % and 41.1 % respectively. Maximalconcentrations achieved were 277 and 184 mg/gbiomass respectively. Fermentation brought to light high inhibition from both native wood and particleboard sources ofmedia, which were attributed to components or degraded products of lignocellulose that were notanalysed in this project. Ethanol was formed during fermentation, with reduced productivity butincreased yields as compared with the control sample. Inhibition was so strong that no difference couldbe given between native and particleboard wood. In this situation, no inhibition potential of resin orits degradation products could be proved. / Lignocellulosic biomassa har potential att bidra till kemikalier och biobränsletillförsel i framtidasamhällen, trots att lignocellulosa är en rekalcitrant struktur som måste behandlas i flera steg. Idagträmaterial som spånskivor bara används för energiåtervinning och värmeproduktion efter deraslivscykel. De kan därför användas som råvara för framställning av värdefulla molekyler.Fermenteringsprocesser behöver frisättningen av trä monosackarider genom förbehandlingsprocesseroch enzymatisk hydrolys. Studien fokuserar på potentialen för avfall från spånskivor för kemisk ochbiobränsleproduktion. Vi har jämfört sackarifiering mellan nativt trä och spånskivor genom simuleradångaxplosion och enzymatisk hydrolys, med en inblick i efterföljande fermentering av Saccharomycescerevisiae. Spånskivor bunden av urea-formaldehyd undersöktes, liksom vissa aspekter av spånskivorbundna med melamin-urea-formaldehyd. Förbehandlingen producerade högre koncentration av lignocellulosa nedbrytningsprodukter frånnativt trä jämfört med spånskivor. Bildningen av nedbrytningsprodukter från sockerarter - furfural och5-hydroxymethylfurfural - ökade med längre förbehandlingar. En väsentlig fraktion av limmet borttogsfrån spånskivorna, vilket ledde till jämförbara koncentrationer i fri urea, dess nedbrytningsprodukteroch formaldehyd mellan naturligt trä och spånskivor under enzymatisk hydrolys. Enzymatisk hydrolys med cellulaser och hemicellulaser avslöjade den kritiska rollen av förbehandlingför att förbättra utbytet, både i naturligt trä och i urea-formaldehyd spånskiva. Längre (20 minuter)ångexplosion vid 160° C resulterade i högre glukosutbytet (från 18,5% till 32,8% för naturligt trä ochfrån 15,6% till 37,4% för spånskivor). Förlängning av uppehållstiden före behandlingen till 35 minuterresulterade i mycket bättre glukosekstraktion för nativt trä (64,5%) men endast liten framsteg förspånskivan (41,1%). Detta resulterade i maximalt utbyte av 277 mg Glc/g biomassa och 184 mg Glc/ gbiomassa för nativt trä och spånskivor, respektive. Fermentering visade hög hämning från lignocellulosa nedbrytningsprodukter som inte analyserades iprojektet för både nativt trä och spånskällor för media. Etanol bildades under fermentering medreducerad produktivitet men ökade utbyten jämfört med kontrollprovet. Hämningen var så stark attingen skillnad kunde ges mellan naturligt trä och spånskivor. I denna situation kunde ingenhämningspotential för lim eller dess nedbrytningsprodukter bevisas.

Tumor Necrosis Factor-Like Ligand 1a (TL1A) is a cytokine encoded by Tumor Necrosis Factor Super Family 15 gene (TNFSF15) gene mostly in endothelial cells which binds to T-cells and foments the production of pro-inflammatory cytokines including TNF-α, IL-6, IL-1b, IFN- γ and IL-13. TL1A level is elevated in inflammatory diseases including Crohn's Disease (CD). Although Single Nucleotide Polymorphisms (SNPs) in TNFSF15 have been reported in CD, no studies have investigated the effect of these SNPs on TL1A, inflammation, and susceptibility to Mycobacterium avium subspecies paratuberculosis (MAP) infection. MAP is a strong candidate in CD pathogenesis. This study is designed to elucidate the combined effect of MAP and SNPs in TNFSF15 (rs4263839, rs7848647, rs6478108, or rs6478109) on TL1A secretion and downstream effect on pro-inflammatory cytokines. Peripheral blood from CD and healthy subjects was analyzed for MAP DNA, TNFSF15 genotyping, circulating TL1A level, and IFN- γ and TNF-α gene expression. Our data is first to report that rs4263839, rs7848647, rs6478108, and rs6478109 in TNFSF15 resulted in increase in circulating TL1A level in healthy and CD samples. Specifically, in CD samples with rs7848647, the average TL1A level was 146.9 pg/mL ± 124.5 compared 62.4 pg/mL ± 82.8 in normal samples. Similarly, TL1A level in CD samples with rs6478109 was 141.9 pg/mL ± 127.7 compared to 71.5 pg/mL ± 88.4 in normal samples (p < 0.05). All 4 SNPs resulted in significant elevation in TL1A level in healthy samples (p < 0.05). Moreover, IFN-γ expression was significantly higher, by approximately 1.6-fold in CD patients with SNPs relative to CD patients with no SNPs (p < 0.05). Interestingly, SNPs in TNFS15 had no significant effect on TNF-α expression. MAP was detected in the blood of 63% of CD compared to 6% healthy subjects (p < .001). The data did not support a correlation between MAP presence and circulating TL1A levels, and no correlation between SNPs in TNSF15 and MAP susceptibility. This study strongly suggests, that SNPs in TNFSF15 increase TL1A levels and may be a contributory factor to the inflammation experienced by CD patients. Over all, the study emphasizes the need for a pharmacogenomic approach in treatment delivery for patients with CD by using TNFSF15 SNPs to identify patients that would benefit from biologics targeting TL1A rather than TNF-α for more efficacious treatment regiments for CD patients.