The role of viral sensing in the development of autoimmunity

Type I diabetes (T1D) is an autoimmune disease caused by a combination of genetic and environmental factors. Previous studies have linked infections with enteroviruses, such as coxsackievirus B4 (CVB4), with an increased risk of T1D. This dissertation examines the impact of both exogenous virus (i.e. CVB4) and endogenous retroviruses (ERVs) residing in the mammalian genome on the development of chronic inflammation and autoimmunity. Following CVB4 infection, T1D-susceptible mice displayed exaggerated tissue damage and signs of chronic inflammation after viral clearance. Ongoing inflammation in these mouse strains was linked to an increased infiltration of T follicular helper cells (Tfh) and germinal centre-like B cells in the pancreas. Furthermore, genome-wide expression analyses of pancreatic beta cells from CVB4-infected mice revealed the activation of virus response genes associated with type I/II interferon (IFN) signalling, accompanied by tissue-specific induction of ERVs. In addition, our data highlight an important role for ERVs in the transcriptional regulation of immune response genes. Remarkably, pancreatic beta cells from uninfected T1D-susceptible mice revealed similar gene enrichment profiles to those found in CVB4-infected mice, emphasizing the role of viral sensing in autoimmune diabetes. To further explore the role of viral recognition in autoimmunity, we made use of a novel mutant mouse strain that carries a point mutation within the MAVS (mitochondrial antiviral-signalling) protein, a crucial mediator of cytoplasmic nucleic acid sensing. MAVSlos mice immunised with RNA substrates exhibited diminished production of type I IFN and subsequent reduction of Tfh expansion and germinal centre formation. These findings demonstrate the significant role of (viral) nucleic acid sensing for appropriate activation of effector cells, but also illustrate how dysregulation of innate anti-viral signalling may translate into inappropriate (diminished or exaggerated) adaptive immunity

In vitro activity of immunosuppressive drugs against Plasmodium falciparum

Background Solid organ transplant recipients are particularly vulnerable for infectious diseases due to prolonged immunosuppressive treatment. Residents of endemic regions and travellers may be exposed to malaria and may, therefore, require prolonged antimalarial chemoprophylaxis. The hypothesis of this study was that certain immunosuppressive drugs may exert clinically relevant anti- malarial activity. It was therefore designed to assess the intrinsic anti- malarial activity of everolimus, mycophenolic acid, and rapamycin against Plasmodium falciparum in an in vitro model. Methods Three laboratory adapted clones of P. falciparum and two isolates were used to assess the potential of mycophenolic acid, rapamycin and everolimus to inhibit in vitro growth of P. falciparum. The standard histidine rich protein 2 assay was employed and inhibitory drug concentrations (IC) were computed by non-linear regression analysis. Results All drugs were associated with complete inhibition of P. falciparum growth in in vitro assays. Mycophenolic acid demonstrated IC50 and IC90 values of 5.4 μmol/L and 15.3 μmol/L. Rapamycin inhibited P. falciparum growth at 7.2 μmol/L (IC50) and 12.5 μmol/L (IC90), respectively. Finally, everolimus displayed IC50 and IC90 values of 6.2 μmol/L and 11.5 μmol/L. There was no difference in in vitro activity against chloroquine sensitive or chloroquine resistant parasites. Conclusions All immunosuppressive drugs evaluated in this in vitro study demonstrated activity against P. falciparum. Inhibitory concentrations of mycophenolic acid are within clinically achievable plasma concentrations when used in solid organ transplant recipients. Further in vivo evaluation of mycophenolic acid either alone or in combination regimens may prove promising for the concomitant prevention of P. falciparum in solid organ transplant recipients living or travelling in malaria endemic regions

Machine-learning-derived classifier predicts absence of persistent pain after breast cancer surgery with high accuracy

Background Prevention of persistent pain following breast cancer surgery, via early identification of patients at high risk, is a clinical need. Supervised machine-learning was used to identify parameters that predict persistence of significant pain. Methods Over 500 demographic, clinical and psychological parameters were acquired up to 6 months after surgery from 1,000 women (aged 28-75 years) who were treated for breast cancer. Pain was assessed using an 11-point numerical rating scale before surgery and at months 1, 6, 12, 24, and 36. The ratings at months 12, 24, and 36 were used to allocate patents to either "persisting pain" or "non-persisting pain" groups. Unsupervised machine learning was applied to map the parameters to these diagnoses. Results A symbolic rule-based classifier tool was created that comprised 21 single or aggregated parameters, including demographic features, psychological and pain-related parameters, forming a questionnaire with "yes/no" items (decision rules). If at least 10 of the 21 rules applied, persisting pain was predicted at a cross-validated accuracy of 86% and a negative predictive value of approximately 95%. Conclusions The present machine-learned analysis showed that, even with a large set of parameters acquired from a large cohort, early identification of these patients is only partly successful. This indicates that more parameters are needed for accurate prediction of persisting pain. However, with the current parameters it is possible, with a certainty of almost 95%, to exclude the possibility of persistent pain developing in a woman being treated for breast cancer.Peer reviewe

The endothelial glycocalyx prefers albumin for evoking shear stress-induced, nitric oxide-mediated coronary dilatation

Background: Shear stress induces coronary dilatation via production of nitric oxide ( NO). This should involve the endothelial glycocalyx ( EG). A greater effect was expected of albumin versus hydroxyethyl starch ( HES) perfusion, because albumin seals coronary leaks more effectively than HES in an EG-dependent way. Methods: Isolated hearts ( guinea pigs) were perfused at constant pressure with Krebs-Henseleit buffer augmented with 1/3 volume 5% human albumin or 6% HES ( 200/0.5 or 450/0.7). Coronary flow was also determined after EG digestion ( heparinase) and with nitro-L-arginine ( NO-L-Ag). Results: Coronary flow ( 9.50 +/- 1.09, 5.10 +/- 0.49, 4.87 +/- 1.19 and 4.15 +/- 0.09 ml/ min/ g for `albumin', `HES 200', `HES 450' and `control', respectively, n = 5-6) did not correlate with perfusate viscosity ( 0.83, 1.02, 1.24 and 0.77 cP, respectively). NO-L-Ag and heparinase diminished dilatation by albumin, but not additively. Alone NO-L-Ag suppressed coronary flow during infusion of HES 450. Electron microscopy revealed a coronary EG of 300 nm, reduced to 20 nm after heparinase. Cultured endothelial cells possessed an EG of 20 nm to begin with. Conclusions: Albumin induces greater endothelial shear stress than HES, despite lower viscosity, provided the EG contains negative groups. HES 450 causes some NO-mediated dilatation via even a rudimentary EG. Cultured endothelial cells express only a rudimentary glycocalyx, limiting their usefulness as a model system. Copyright (c) 2007 S. Karger AG, Basel

Temperature Effects on Biomass and Regeneration of Vegetation in a Geothermal Area.

Understanding the effects of increasing temperature is central in explaining the effects of climate change on vegetation. Here, we investigate how warming affects vegetation regeneration and root biomass and if there is an interactive effect of warming with other environmental variables. We also examine if geothermal warming effects on vegetation regeneration and root biomass can be used in climate change experiments. Monitoring plots were arranged in a grid across the study area to cover a range of soil temperatures. The plots were cleared of vegetation and root-free ingrowth cores were installed to assess above and below-ground regeneration rates. Temperature sensors were buried in the plots for continued soil temperature monitoring. Soil moisture, pH, and soil chemistry of the plots were also recorded. Data were analyzed using least absolute shrinkage and selection operator and linear regression to identify the environmental variable with the greatest influence on vegetation regeneration and root biomass. There was lower root biomass and slower vegetation regeneration in high temperature plots. Soil temperature was positively correlated with soil moisture and negatively correlated with soil pH. Iron and sulfate were present in the soil in the highest quantities compared to other measured soil chemicals and had a strong positive relationship with soil temperature. Our findings suggest that soil temperature had a major impact on root biomass and vegetation regeneration. In geothermal fields, vegetation establishment and growth can be restricted by low soil moisture, low soil pH, and an imbalance in soil chemistry. The correlation between soil moisture, pH, chemistry, and plant regeneration was chiefly driven by soil temperature. Soil temperature was negatively correlated to the distance from the geothermal features. Apart from characterizing plant regeneration on geothermal soils, this study further demonstrates a novel approach to global warming experiments, which could be particularly useful in low heat flow geothermal systems that more realistically mimic soil warming