The O’Brien lab has demonstrated that Heat shock protein 27 (HSP27)shows attenuated expression in human coronary arteries as the degree of atherosclerosis progresses. Moreover, over-expression of HSP27 reduces atherogenesis in mice. The precise mechanism(s) for HSP27-mediated "atheroprotection" are incompletely understood. Nuclear Factor-kappaB (NF-κB) is a key signaling modulator in atherogenesis. Hence, this project sought to determine if recombinant HSP27 (rHSP27) alters NF-κB signaling to affect atheroprotection. Treatment of THP1 macrophages with rHSP27 resulted in degradation of IκBα, coincided with nuclear translocation of the p65 subunit and produced transcriptional evidence of activation of NF-κB signaling. When the transcriptional profile of THP1 macrophages treated with rHSP27 was analyzed using NF-κB-pathway-specific qRT-PCR arrays, among the regulated genes, IL-10 and GM-CSF mRNA levels were markedly increased, as were parallel translational effects observed. These data provide new mechanistic insights into the atheroprotective effects of HSP27.

No description available.

El trasplantament d'illots pancreàtics és una teràpia emergent per la curació de la diabetis mellitus. Una de les limitacions radica en la baixa disponibilitat d'òrgans i l'elevada demanda existent, que queda agreujada amb l'elevat nombre d'illots que són necessaris per restablir la normoglucèmia del pacient. Estudis recents del nostre grup, han mostrat que en els primers dies després del trasplantament hi ha un augment de l'expressió d'IL-1beta en els empelts d'illots.La hipòtesi de treball és que la citocina proinflamatòria, IL-1, està implicada en la fallada del trasplantament. Designant la sobreexpressió d'IL-1Ra com l'estratègia a seguir per millorar el pronòstic del trasplantament singènic d'illots pancreàtics. Per tant, l'objectiu general de l'estudi va ser determinar si la sobreexpressió d'IL-1Ra en els illots pancreàtics protegeix les cèl·lules beta pancreàtiques dels efectes deleteris d'IL-1 en els illots i millora el pronòstic del trasplantament.L'estudi dels efectes d'IL-1beta i de la sobreexpressió d'IL-1Ra in vitro es va realitzar amb un cultiu primari d'illots de rata que van ser exposats durant 48h a 5.5 o 22.2 mM de glucosa en presència o absència de 50U/ml d'IL-1beta. I la inserció del gen exogen a les cèl·lules dels illots es va fer utilitzant un adenovirus V recombinant.La proliferació de les cèl·lules beta (determinada per incorporació de BrdU) va disminuir dràsticament quan es van exposar els illots a 50 U/ml d'IL-1beta, tant a 5.5 mM com a 22.2 mM de glucosa. Aquest efecte d'IL-1beta va quedar completament abolit per la sobreexpressió d'IL-1Ra en els illots que havien estat infectats amb l'adenovirus que codificava per l'antagonista, a les dues concentracions de glucosa utilitzades.L'apoptosi de les cèl·lules beta (determinada per immunohistoquímica mitjançant la tècnica del TUNEL i per citometria de flux, marcant les cèl·lules amb anexina V i iodur de propidi) estava significativament augmentada en els illots exposats a IL-1beta, però no en els illots que sobreexpressaven IL-1Ra.L'estudi dels efectes de la sobreexpressió d'IL-1Ra en els illots trasplantats es va realitzar utilitzant un model de trasplantament singènic. Grups de 500 illots control (no infectats) o que sobreexpressaven IL-1Ra van ser trasplantats sota la càpsula renal de rates Lewis diabètiques. 500 illots són una massa beta clarament insuficient per restablir la normoglucèmia, així doncs els animals d'ambdós grups es van mantenir hiperglucèmics durant tot l'estudi. Els empelts es van recuperar després de 3, 10 i 28 dies del trasplantament i es van processar per fer estudis histològics.La sobreexpressió d'IL-1Ra en els illots trasplantats va fer augmentar significativament la proliferació de les cèl·lules beta dels empelts de 3, 10 i 28 dies i va protegir parcialment les cèl·lules beta de l'increment d'apoptosi detectat després del trasplantament, tant a curt com a llarg termini. L'àrea individual de les cèl·lules beta estava augmentada de manera similar tant en els empelts d'illots control com en els illots que sobreexpressaven IL-1Ra als 10 i 28 dies d'evolució. Finalment, la sobreexpressió d'IL-1Ra resultà en una recuperació de la massa beta inicialment trasplantada.Per tal d'estudiar si els efectes beneficiosos de la sobreexpressió d'IL-1Ra aconseguien reduir el nombre d'illots necessaris per restablir la normoglucèmia, es va trasplantar una massa beta marginal (800 illots) d'illots control i Ad-IL-1Ra a animals diabètics. El 100% dels animals trasplantats amb illots Ad-IL-1Ra eren normoglucèmics després de 14 dies del trasplantament i només un 40% dels animals trasplantats amb illots control assoliren l'euglucèmia en aquest dia.En aquest treball es mostra que la citocina proinflamatòria IL-1beta indueix clarament apoptosi a les cèl·lules beta dels illots de rata en cultiu i inhibeix dràsticament la replicació d'aquestes cèl·lules. La sobreexpressió d'IL-1Ra protegeix les cèl·lules beta dels efectes deleteris d'aquesta citocina i amplifica la resposta replicativa de les cèl·lules beta exposades a concentracions altes de glucosa. La sobreexpressió d'IL-1Ra en els illots augmenta la replicació de les cèl·lules beta trasplantades, les protegeix de l'apoptosi induïda després del trasplantament, i preserva la massa beta inicialment trasplantada. Els efectes beneficiosos de la sobreexpressió d'IL-1Ra observats en els illots trasplantats permeten reduir el nombre d'illots necessaris per restablir la normoglucèmia dels animals diabètics.Aquests resultats suggereixen que la IL-1 juga un paper important en l'evolució dels empelts d'illots, ja que el seu bloqueig implica una millora dels illots trasplantats. / BACKGROUND AND AIMS: IL-1beta could contribute to the dramatic beta cell loss that takes place after islet transplantation. It is known that exposure to sustained hyperglycemia has a deleterious effect on transplanted islets. Moreover, it has been recently reported that IL-1beta expression is increased in islets exposed to high glucose levels. IL-1Ra is a naturally occurring inhibitor of IL-1 action and its overexpression protects pancreatic islets from the deleterious effects of IL-1â on beta cell replication, apoptosis and function. The aim of this study was to determine whether viral gene transfer of the IL-1Ra gene into rat islets ex vivo could have a beneficial effect on beta cell replication and mass of transplanted islets.METHODS:Lewis rat islets were infected for 2h with 6.25 × 106 pfu of Ad-IL-1Ra and streptozotocin-diabetic Lewis rats were transplanted with 500 Ad-IL-1Ra infected islets (Ad-IL-1Ra group) or 500 uninfected islets (control group) under the kidney capsule. Grafts were removed 3 (n = 12), 10 (n = 12) and 28 (n = 12) days after transplantation and beta cell replication, apoptosis and mass were determined.RESULTS:500 islets is an insufficient mass to restore normoglycemia and therefore, all animals but one (IL-1Ra group) remained hyperglycemic until the end of the study. Beta cell replication (determined by BrdU incorporation) was significantly increased in Ad-IL-1Ra group on days 3 (0.78 ± 0.23%), 10 (1.15 ± 0.16%) and 28 (1.22 ± 0.2%) after islet transplantation compared to beta cell replication in normal pancreas (0.24 ± 0.04%; p< 0.05). In contrast, in control group, beta cell replication was not increased on day 3 after transplantation (0.41 ± 0.11%), and although it increased on day 10 (0.89 ± 0.18%; p< 0.01) it was reduced again on day 28 (0.59 ± 0.10%) in agreement with previous reports of limited beta cell replication with persistent hyperglycemia. Beta cell apoptosis (determined by TUNEL method) was significantly increased in transplanted islets from both groups compared to pancreas. Although Ad-IL-1Ra group showed lower beta cell apoptotic levels than control group, differences did not reach statistical significance. The initially transplanted â-cell mass (1.34 ± 0.03 mg) was similarly reduced in both control (0.32 ± 0.06 mg) and Ad-IL-1Ra groups (0.45 ± 0.10 mg) (p<0.001) on day 3 after transplantation. In Ad-IL-1Ra islet grafts, beta cell mass increased after 10 (1.04 ± 0.091 mg; p< 0.010) and 28 (0.8 ± 0.24 mg) days of transplantation. In contrast, beta cell mass of control group was also increased on day 10 after transplantation (0.69 ± 0.12 mg), but it dropped again on day 28 (0.41 ± 0.05 mg) paralleling with the evolution of beta cell replication in this group. CONCLUSIONS:Islets overexpressing IL-1Ra showed an increased beta cell replication and a preserved beta cell mass after transplantation, that was maintained even after longterm exposure to hyperglycemia.

B cells can be activated by T cell-dependent stimuli, such as CD40 ligation and cytokines, which induce extensive proliferation, class switch recombination and somatic hypermutation. Epstein-Barr virus (EBV) can also induce B cell activation by mimicking T cell help through its main oncoprotein, latent membrane protein 1 (LMP-1). It is regulated by another EBV-encoded protein, EBV nuclear antigen 2 (EBNA-2), which is absent in Hodgkin and Burkitt lymphomas. We have studied LMP-1 induction by cytokines in vitro and shown that LMP-1 is induced through the transcription factor signal transducer and activator of transcription (STAT6) and a newly defined high-affinity STAT6-binding site. When IL-4 is added together with lipopolysaccharide (LPS) or α-CD40 to B cells, it induces homotypic round and tight aggregates in vitro, whereas LPS alone does not induce such morphological changes. I describe here attempts to identify the molecules that regulate these responses. I have shown that the Rho GTPase Cdc42 controls the spreading of B cells, whereas two other molecules in the same family, Rac1 and Rac2, control homotypic adhesion. Further, I have shown by conditional deletion of Cdc42 in B cells that it is important in the humoral immune response.  Dock10 is a guanosine nucleotide exchange factor (GEF) for Cdc42. It is expressed through all differentiation stages of B cell development. However, targeted deletion of Dock10 in B cells does not result in an aberrant phenotype. Furthermore, by studying conditional knockout mice for Dock10, Cdc42, Rac1 and Rac2, I have elucidated the mechanism of cytoskeletal changes during B cell activation, leading to adhesion and motility. My results may lead to a better understanding of normal B cell activation and of EBV infection, which is associated with many human tumours and may help to understand cancer development and progression in B cells. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Manuscript. Paper 3: Manuscript.</p>

In 1984 the Swedish government introduced a special law which gave tax benefits to foreign researchers who for a limited time was engaged in research or development work in Sweden. The purpose was to facilitate for companies to recruit skilled employees from outside of Sweden and to make Sweden more attractive to this important group of researchers and to strengthen the Swedish competitiveness. In 2001 the law for foreign researchers was reformed to include foreign experts, researchers, managers and other key personnel. In Sweden called “expert tax”. As a result of this; problem with the predictability of who may have the potential to apply for the expert tax benefits was developed. This thesis deal with the tax rules for foreign expert, focusing on the predictability in order to apply for the expert tax. The focus has been on analyse the case law in this area to see if the case law has expanded the tax purposes or if the problem lies in the tax basic design in terms of predictability. Problems today are found in the preambles, case law and doctrine. The law and preambles for the expert tax rules have a vague formulation. Hence, this is why so many interpretations have been done by Foskarskattenämnden and the Swedish courts. This has led to the purpose of the tax being enlarged and do not exclude any business or employees today. This is why it is so difficult to predict who would become suitable for the expert tax, which indicates that the tax is vague formulated and not precise enough. The complexity has lead to interpreting the law in each case which may produce results that can differ among cases of similar nature. We need stronger legislation and a more clear picture of what the tax rules for foreign expert stand for, in order to obtain a tax functioning and avoid predictability problems.

In the present study our objectives were (1) to follow the temporal patterns of testicular LH and FSH receptor (LH-R and FSH-R) concentrations and affinity (Ka) during sexual maturation in bulls, to see if such patterns could explain the control of rapid testicular growth that occurs after 25 weeks of age, when serum gonadotropin concentrations are low; (2) to see if transformation growth factors (TGF- alpha and beta 1, 2 and 3) and interleukins (IL-1 and IL-6) are produced in the developing bovine testis and if their concentrations change during development; (3) to see if the onset of puberty could be hastened by treating bull calves subcutaneously (sc) with 3 mg of bLH (n=6) or 4 mg of bFSH (n=6) once every 2 days, from 4 to 8 weeks after birth. Mean LH-R concentrations decreased from 13 to 25 weeks of age and increased to 56 weeks of age (P<0.05). LH-RKa decreased from 9 to 17 weeks of age, increased to 29 weeks and declined to 33 weeks of age (P<0.05). FSH-R concentrations declined from 17 to 25 weeks of age then increased to 56 weeks of age (P<0.05). FSH-RKa increased from 17 to 25 weeks of age (P<0.05). Testicular TGF-alpha concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks and from 33 to 56 weeks of age (P<0.05). Testicular TGF-beta 1 concentrations decreased from 17 to 21 weeks of age, increased to 25 weeks and decreased from 25 to 29 weeks of age (P<0.05). Testicular TGF-beta 2 concentrations increased from 5 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks and decreased at 29 weeks of age (P<0.05). Testicular TGF-beta 3 concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks of age and from 25 to 29 weeks of age (P<0.05). Mean testicular IL-1 alpha concentrations decreased from 5 to 9 weeks of age and 13 to 21 weeks of age (P<0.01) while mean testicular IL-1 beta concentrations decreased from 13 to 17 weeks and 29 to 33 weeks of age (P<0.01). Mean IL-1 bioactivity increased from 13 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks, decreased to 29 weeks and decreased from 33 to 56 weeks of age (P<0.05). Mean testicular IL-6 concentrations decreased (P<0.05) from 9 to 13 weeks of age, increased (P<0.05) to 21 weeks, decreased (P<0.05) to 25 weeks of age, increased (P<0.05) to 29 weeks and decreased (P<0.01) to 56 weeks of age. <p>We concluded that high concentrations of gonadotropin receptors might be critical to initiate postnatal testis growth and support it after 25 weeks of age in the face of low serum gonadotropin concentrations. Testicular TGF-alpha concentrations were higher in calves than adults while concentrations of TGF-beta and IL-1 were higher in the early postnatal period than the peripubertal period. The changes in testicular concentrations of TGFs and ILs led us to suggest a possible local regulatory role in development. Testicular IL-6 concentrations were higher in prepubertal calves than adults. Treatment of bull calves with bFSH from 4 to 8 weeks of age increased testicular growth (SC), hastened the onset of puberty (SC ≥ 28 cm), and enhanced spermatogenesis.

Avsikten med min uppsats är att analysera Levinas begrepp Il y a, det finns, som han utvecklar som en del av sin kritik mot Heideggers fundamentalontologi. Levinas diskussion om det finns är riktad mot Heideggers idé att ångesten utsätter oss för intet, något som Levinas motsätter sig eftersom han menar att man aldrig kan nå intet i livet. Den centrala frågeställningen i denna uppsats är varför det finns är ett nödvändigt begrepp, vi har ju redan Heideggers begrepp varat? I min uppsats jämför jag även Levinas idé om det finns med Heideggers idé om långtråkigheten, ångesten och kastadheten. I sitt letande efter varat utforskar Heidegger två sinnestillstånd: det ena, långtråkigheten, uppenbarar varat. För Heidegger är långtråkigheten en stämning bland andra möjliga, där varat som sådant framträder för oss just för att det varande drar sig tillbaka, på ett liknande sätt som i ångesten, den motsatta sidan av samma mynt. I den uppenbaras intet. Mot detta ställer Levinas en annan upplevelse: sömnlösheten, som synliggör det han kallar det finns.

The story of Joseph (in the Bible), Yusuf (in the Quran), has inspired literatures in many languages. This paper explores how some Swahili writers and translators have dealt with this inspiration, the implications for their language use and the way they have interpreted Yusuf as a theme for their writings. After a brief introduction on the importance of the story itself and putting the focus on a major theme of the plot, the following works are discussed: the new Quran translation by Sh Ali Muhsin (1995), a short novel by Mzee Salim A. Kibao (1975), two short stories by Amur bin Nasur il-Omeiri (1894), the utenzi Qissat-il Yusuf (l913) and Abdulrazak Gurnah\'s English written novel Paradise (1995). The paper concludes with the observation that in analyzing how these Swahili writers have integrated the story of Yusuf in their writings, prose as well as poetry, it becomes clear that attempts in defining what is ‘foreign’ (or ‘Oriental’) and what is ‘indigenous’ (or ‘African’) are bound to fail.

<p>The role of two cell extrinsic signals, T cell receptor (TCR) ligation and interleukin-7, in promoting glucose uptake and survival of T lymphocytes is examined in this work. Both of these signals are capable of regulating the uptake and fate of glucose, but the requirement of this regulation for T cell homeostasis and functionality remains unclear. To examine the role of TCR mediated increases of glucose metabolism and the signals involved, primary murine T cells were activated in vitro and the role and regulation of glucose uptake was examined. We show that glucose uptake is limiting in T cell activation and that CD28 costimulation is required for maximal glucose uptake following TCR stimulation by upregulating expression and promoting the cell surface trafficking of the glucose transporter Glut1. Regulation of T cell glucose uptake and Glut1 was critical, as low glucose prevented appropriate T cell responses. Additionally, transgenic expression of Glut1 augmented T cell activation, and led to accumulation of readily activated memory-phenotype T cells with signs of autoimmunity in aged mice. To further examine the regulation of glucose uptake, we analyzed CD28 activation of Akt, which appeared necessary for maximal glucose uptake of stimulated cells and which we have shown can promote Glut1 cell surface trafficking. Consistent with a role for Akt in Glut1 trafficking, transgenic expression of constitutively active Akt (mAkt) increased glucose uptake of resting T cells, but did not alter Glut1 protein levels. Therefore, CD28 appeared to promote Akt-independent upregulation of Glut1 protein and Akt-dependent Glut1 cell surface trafficking. In support of this model, co-expression of Glut1 and mAkt transgenes resulted in a synergistic increase in glucose uptake and accumulation of activated T cells in vivo that were largely independent of CD28. Induction of Glut1 protein and Akt regulation of Glut1 trafficking are therefore separable functions of CD28 costimulation that cooperate to promote glucose metabolism necessary for T cell activation and proliferation.</p><p>Glucose uptake is dramatically increased in response to TCR and costimulation signaling, however, glucose uptake must be maintained at a low level in naive T cells to promote survival and homeostasis. Interleukin-7 (IL-7) plays a central role in maintaining naive T cell homeostasis, and mediates this effect in vivo at least in part through control of homeostatic proliferation and inhibition of apoptosis. IL-7 can promote glucose uptake and glycolysis in vitro and may also promote glucose metabolism in vivo to maintain T cell survival. To determine if IL-7 regulates T cell metabolism in vivo, we generated a transgenic model for conditional IL-7 receptor (IL 7R) expression on IL-7R-/- T cells. T cells in this model developed normally and, consistent with previous work, deletion of the IL-7R transgene in vivo led to cell death even in an otherwise normal lymphoid compartment. Importantly, in vivo deletion of IL 7R also led to decreased cell size and glycolytic flux. However, glucose uptake was not altered following deletion of the IL-7R indicating that while not essential for glucose uptake, IL-7 is required for maintenance of glycolysis. These data are the first to identify a signal required in vivo to regulate lymphocyte metabolism and demonstrate that in addition to its well-defined roles in homeostatic proliferation and cell survival, IL-7 plays a key and non-redundant role to maintain T cell glycolysis. Together, these data concerning the role of TCR, costimulation, and IL-7 in the regulation of glucose uptake and metabolism exemplify the importance of cell extrinsic signals and the regulation of glucose utilization.</p> / Dissertation

<p>Selenium (Se) is an essential trace element and metalloid involved in several key metabolic activities: protection against oxidative damage, regulation of immune and thyroid function, and fertility. Several recent lines of evidence from epidemiology, genetic, and transgenic animal studies suggest that Se may play a protective role in Osteoarthritis (OA). However, the exact protective mechanism of Se is still unclear. </p><p>In this study, we hypothesized that Se exerts its chondroprotective benefit via an anti-oxidative and anti-inflammatory effect mediated by specific selenoproteins that neutralize cytokine-induced inflammatory responses in chondrocytes. We established an in vitro system for studying the effect of Se in the chondrosarcoma cell line SW-1353 and in human primary chondrocytes. Selenomethionine (SeMet) induced gene expression and enzyme activity of both antioxidative enzymes glutathione peroxidase (GPX) and thioredoxin reductase (TR) in SW-1353 cells. Our data suggest that Se may be protective against oxidative stress through regulation of the activity of these antioxidative enzymes.</p><p>As IL-1&beta; is one of the primary pro-inflammatory cytokines contributing to the progression in OA, we next investigated the effect of Se on the gene expression induced by physiological doses of IL-1&beta;. SeMet inhibited IL-1&beta; induced catabolic gene expression of matrix metalloproteinase 1 (MMP1) and MMP13 as well as total MMP activity in chondrocytes. Similarly, SeMet inhibited chondrocyte gene expression of IL-1&beta; induced nitric oxide synthase (iNOS) and cyclooxygenase (COX2) with corresponding reductions in nitric oxide (NO) and prostaglandin E2 (PGE2) production. In addition, SeMet pretreatment attenuated the IL-1&beta; induced activation of p38 MAPK but not the ERK, JNK or NFkB pathways. Taken together, our results suggest that Se inhibits IL-1&beta; induced expression of inflammatory and catabolic genes, partly through inhibition of IL-1&beta; cell signaling. </p><p>Since Se may function through selenoproteins, we evaluated the role of three specific major selenoproteins, GPX1, TR1 and DIO2, in modifying the inflammatory response stimulated by IL-1&beta; in chondrocytes by RNA interference. Based on RNA interference results, DIO2 and TR1 mediated the inhibitory effect of SeMet on IL-1&beta; induced COX2 gene expression, while GPX1 did not show a significant inhibitory effect on Se. Depletion of DIO2 increased the IL-1&beta; induced COX2 gene expression. This suggests that DIO2 may negatively modulate the IL-1&beta; response. Our data also suggest that part of this inhibitory effect of DIO2 could be through regulation of IL-1&beta; gene expression itself. These results highlight a potential new role of DIO2 in modulating the inflammatory response in chondrocytes </p><p>In summary, the result of this study suggests that Se may exert its chondroprotective effect through specific selenoproteins which neutralize oxidative stress and modify the inflammatory response in chondrocytes.</p> / Dissertation