Genetic variants in the NOD2/CARD15 gene are associated with early mortality in sepsis patients

Objective: Genetic variants in the NOD2/CARD15 gene resulting in adiminished capacity to activate NF-κB in response to bacterial cell wall products have been associated with Crohn's disease (CD). Recently, we found an association between the variant Leu1007fsinsC of the NOD2/CARD15 gene (SNP13) and asignificantly increased rate of transplant related mortality (TRM) due to intestinal and pulmonary complications in stem cell transplantation (SCT). To assess apossible contribution of variants in the NOD2/CARD15 gene to sepsis related mortality (SRM) we investigated 132 prospectively characterised, consecutive patients with sepsis. Design and patients: The three most common NOD2/CARD15 variants (Arg702Trp, Gly908Arg, and Leu1007fsinsC) were determined in 132 prospectively characterised patients with sepsis attended to three intensive care units at the University of Regensburg by Taqman PCR. NOD2/CARD15 genotype and major patients' characteristics were correlated with SRM. Results: Patient groups with and without NOD2/CARD15 variants did not differ in their clinical characteristics such as median age, gender, reason for admission or APACHE score; however, SRM (day30) was increased in patients with NOD2/CARD15 coding variants (42 vs. 31%) and was highest (57%) in 8 patients carrying the Leu1007fsinsC variant (p < 0.05). Multivariate analysis demonstrated the Leu1007fsinsC genetic variant as an independent risk factor for SRM. Conclusion: Our findings indicate amajor role of NOD2/CARD15 coding variants for SRM. This may be indicative for arole of impaired barrier function and bacterial translocation in the pathophysiology of early sepsis related deat

Systemic Effects by Intrathecal Administration of Triamcinolone Acetonide in Patients With Multiple Sclerosis

In patients suffering from multiple sclerosis (MS), intrathecal injection of triamcinolone acetonide (TCA) has been shown to improve symptoms of spasticity. Although repeated intrathecal injection of TCA has been used in a number of studies in late-stage MS patients with spinal cord involvement, no clinical-chemical data are available on the distribution of TCA in cerebrospinal fluid (CSF) or serum. Moreover, the effects of intrathecal TCA administration on the concentrations of endogenous steroids remain poorly understood. Therefore, we have quantified TCA and selected endogenous steroids in CSF and serum of TCA-treated MS patients suffering from spasticity. Concentrations of steroids were quantified by LC-MS, ELISA, or ECLIA and compared with the blood-brain barrier status, diagnosed with the Reibergram. The concentration of TCA in CSF significantly increased during each treatment cycle up to >5 mu g/ml both in male and female patients (p30 ng/ml (p< 0.001) and severely depressed serum levels of cortisol and corticosterone (p< 0.001). In addition, concentrations of circulating estrogen were significantly suppressed (p< 0.001). Due to the potent suppressive effects of TCA on steroid hormone concentrations both in the brain and in the periphery, we recommend careful surveillance of adrenal function following repeated intrathecal TCA injections in MS patients

Dissociation of somatic growth, time of sexual maturity, and life expectancy by overexpression of an RGD-deficient IGFBP-2 variant in female transgenic mice

Impaired growth is often associated with an extension of lifespan. However, the negative correlation between somatic growth and life expectancy is only true within, but not between, species. This can be observed because smaller species have, as a rule, a shorter lifespan than larger species. In insects and worms, reduced reproductive development and increased fat storage are associated with prolonged lifespan. However, in mammals the relationship between the dynamics of reproductive development, fat metabolism, growth rate, and lifespan are less clear. To address this point, female transgenic mice that were overexpressing similar levels of either intact (D-mice) or mutant insulin-like growth factor-binding protein-2 (IGFBP-2) lacking the Arg-Gly-Asp (RGD) motif (E-mice) were investigated. Both lines of transgenic mice exhibited a similar degree of growth impairment (-9% and -10%) in comparison with wild-type controls (C-mice). While in D-mice, sexual maturation was found to be delayed and life expectancy was significantly increased in comparison with C-mice, these parameters were unaltered in E-mice in spite of their reduced growth rate. These observations indicate that the RGD-domain has a major influence on the pleiotropic effects of IGFBP-2 and suggest that somatic growth and time of sexual maturity or somatic growth and life expectancy are less closely related than thought previously

Metabolic Pathway Modeling in Muscle of Male Marathon Mice (DUhTP) and Controls (DUC)-A Possible Role of Lactate Dehydrogenase in Metabolic Flexibility

In contracting muscles, carbohydrates and fatty acids serve as energy substrates; the predominant utilization depends on the workload. Here, we investigated the contribution of non-mitochondrial and mitochondrial metabolic pathways in response to repeated training in a polygenic, paternally selected marathon mouse model (DUhTP), characterized by exceptional running performance and an unselected control (DUC), with both lines descended from the same genetic background. Both lines underwent three weeks of high-speed treadmill training or were sedentary. Both lines' muscles and plasma were analyzed. Muscle RNA was sequenced, and KEGG pathway analysis was performed. Analyses of muscle revealed no significant selection-related differences in muscle structure. However, in response to physical exercise, glucose and fatty acid oxidation were stimulated, lactate dehydrogenase activity was reduced, and lactate formation was inhibited in the marathon mice compared with trained control mice. The lack of lactate formation in response to exercise appears to be associated with increased lipid mobilization from peripheral adipose tissue in DUhTP mice, suggesting a specific benefit of lactate avoidance. Thus, results from the analysis of muscle metabolism in born marathon mice, shaped by 35 years (140 generations) of phenotype selection for superior running performance, suggest increased metabolic flexibility in male marathon mice toward lipid catabolism regulated by lactate dehydrogenase.Peer reviewe

Bedeutung von Fibronektin Spleißvarianten für die Migration primärer Colon lamina propria Fibroblasten bei unterschiedlichen klinischen Subtypen von Morbus Crohn

Die Migration von Colon Lamina Propria Fibroblasten und die Wundkontraktion spielen bei der Wundheilung und im Verlauf der Entzündung, Strikturen- und Fistelbildung bei chronisch entzündlichen Darmerkrankungen eine wesentliche Rolle. Es ist aus Vorarbeiten unserer Arbeitsgruppe bekannt, dass die Fibroblasten aus der Mukosa von gesunden Kontrollen und Patienten mit unterschiedlichen Subtypen von Morbus Crohn unterschiedlich stark migrieren. Fibroblasten aus entzündeter Mukosa von Patienten mit Morbus Crohn migrieren weniger als Fibroblasten, die aus der gesunden Mukosa von Kontrollpatienten isoliert wurden. Fibroblasten aus Stenosearealen weisen ein höheres und Fibroblasten aus Fistelgewebe von Morbus Crohn-Patienten ein vermindertes Migrationspotential verglichen mit einfach entzündeten Morbus Crohn-Fibroblasten auf. Fibronektin und im Besonderen die Spleißvariante Fibronektin ED-A sind als die induzierenden Faktoren der Fibroblastenmigration bekannt. Des Weiteren ist durch vorangegangene Migrationsassays mit Kontrollfibroblasten, die für 3 Tage mit Interferon-gamma (IFN-gamma) und Tumor Nekrose Faktor (TNF) inkubiert wurden, eine dosisabhängige signifikante Hemmung der Migration beobachtet worden. Bei einer Langzeitinkubation von 6 Tagen mit Transforming Growth Factor-beta1 (TGF-beta1) zeigen intestinale Kontrollfibroblasten ein reduziertes Migrationspotential, während eine 6 stündige Inkubation der Kontrollfibroblasten mit TGF-beta1 zu einer dosisabhängigen Steigerung der Zellmigration führt. Wird ein Kontrollfibroblasten-Monolayer nach einer 6 tägigen Inkubation mit TGF-beta1 mit einem Kamm verwundet und anschließend für 4 Stunden mit dem Wachstumsfaktor Platelet Derived Growth Factor (PDGF) inkubiert, weisen die vorstimulierten Zellen neben der verminderten Migration auch eine reduzierte Focal Adhesion Kinase (FAK)-Autophosphorylierung auf, während die Zellen ohne Vorinkubation eine mit erhöhter FAK-Phosphorylierung verbundene Migrationserhöhung zeigen. In der vorliegenden Arbeit ist es gelungen, die Zusammenhänge zwischen der Migration von Fibroblasten und der Fibronektin Spleißformenexpression zu beleuchten. In Fibroblasten von Patienten mit chronisch entzündlichen Darmerkrankungen korrelierte das Migrationsverhalten mit der Fibronektinexpression. Durch Hemmung bzw. Stimulation der Zellmigration mit den Zytokinen IFN-gamma und TNF bzw. den Wachstumsfaktor TGF-beta1 konnte eine jeweilige verminderte bzw. gesteigerte Fibronektinexpression erzielt werden. Auch der biphasige Effekt von TGF-beta1 konnte auf Fibronektin mRNA-Ebene gezeigt werden. Mit TGF-beta1 vorstimulierte und dadurch zu Myofibroblasten differenzierte Fibroblasten zeigten nach Verwundung und Inkubation mit PDGF eine gesteigerte Fibronektinexpression, die auf eine gesteigerte Synthese der Extrazellulärmatrix zurückzuführen ist, da eine erhöhte Migration aufgrund der verminderten FAK-Autophosphorylierung nicht möglich war. Nicht vorinkubierte Zellen wiesen eine mit der bekannten Migrationserhöhung korrelierende gesteigerte Fibronektinexpression und FAK-Phosphorylierung auf. Weitere Studien zur Synthese der Extrazellulärmatrix, die indirekt durch TGF-beta1 und PDGF induziert wird, könnten zusätzliche Erkenntnisse hinsichtlich der Bildung von Stenosen bei Morbus Crohn geben

Cytokines in milk and the role of TGF-beta

Cytokines are required for normal growth and development of the mammary gland and TGF-β prominently represents an established effector of apoptosis, e.g., during involution of the mammary gland. By the control of intracellular signaling pathways, including JAK/STAT, MAPK, PI-3K, and NF-κB, cytokines efficiently regulate cell proliferation and inflammation in the breast. Therefore, cytokines are discussed also in a context of malignant mammary growth. As a group of tissue hormones produced by somatic cells or by cells from the immune system, cytokines are defined by their immunomodulatory potential. Over the past 40 years, multiple cytokines were identified in colostrum and milk. Importantly, cytokines derived from mammary secretions after birth are required for maturation of the immune system in the developing gastrointestinal tract from the suckling. Moreover, recent studies have further assessed the particular interactions between probiotic bacterial strains and cytokines. In light of the increasing prevalence of inflammatory diseases of the gastrointestinal system, the effects of probiotic microorganisms during milk fermentation may have immunotherapeutic potential in the future

Analysis of Activity-Dependent Energy Metabolism in Mice Reveals Regulation of Mitochondrial Fission and Fusion mRNA by Voluntary Physical Exercise in Subcutaneous Fat from Male Marathon Mice (DUhTP)

Physical inactivity is considered as one of the main causes of obesity in modern civilizations, and it has been demonstrated that resistance training programs can be used to reduce fat mass. The effects of voluntary exercise on energy metabolism are less clear in adipose tissue. Therefore, the effects of three different voluntary exercise programs on the control of energy metabolism in subcutaneous fat were tested in two different mouse lines. In a cross-over study design, male mice were kept for three or six weeks in the presence or absence of running wheels. For the experiment, mice with increased running capacity (DUhTP) were used and compared to controls (DUC). Body and organ weight, feed intake, and voluntary running wheel activity were recorded. In subcutaneous fat, gene expression of browning markers and mitochondrial energy metabolism were analyzed. Exercise increased heart weight in control mice (p &lt; 0.05) but significantly decreased subcutaneous, epididymal, perinephric, and brown fat mass in both genetic groups (p &lt; 0.05). Gene expression analysis revealed higher expression of browning markers and individual complex subunits present in the electron transport chain in subcutaneous fat of DUhTP mice compared to controls (DUC; p &lt; 0.01), independent of physical activity. While in control mice, voluntary exercise had no effect on markers of mitochondrial fission or fusion, in DUhTP mice, reduced mitochondrial DNA, transcription factor Nrf1, fission- (Dnm1), and fusion-relevant transcripts (Mfn1 and 2) were observed in response to voluntary physical activity (p &lt; 0.05). Our findings indicate that the superior running abilities in DUhTP mice, on one hand, are connected to elevated expression of genetic markers for browning and oxidative phosphorylation in subcutaneous fat. In subcutaneous fat from DUhTP but not in unselected control mice, we further demonstrate reduced expression of genes for mitochondrial fission and fusion in response to voluntary physical activity