Inflammatory cytokines influence fatty acid metabolism in hepatocytes

The metabolic syndrome and obesity are currently reaching pandemic dimensions worldwide. The hepatic consequence is an imbalance in fatty acid (FA) homeostasis, which results in hepatic lipid accumulation, a critical characteristic of non-alcoholic fatty liver disease (NAFLD), the most common chronic liver disease. NAFLD is a highly heterogeneous liver disease that ranges from simple steatosis (non-alcoholic fatty liver, NAFL), to non-alcoholic steatohepatitis (NASH), which is associated with inflammation and liver injury, and can ultimately lead to NASH-driven hepatocellular carcinoma (HCC). Pro-inflammatory cytokines, derived from activated immune cells, strongly affect hepatic FA metabolism, and are implicated in NASH and NASH-derived HCC. However, underlying molecular mechanisms and signalling pathways are not known in detail yet. To investigate the influence of NASH-derived inflammatory cytokines on hepatic FA metabolism and to study molecular mechanisms as well as involved signalling pathways, I established an in vitro NASH model to recapitulate the transition from steatosis to steatohepatitis in mouse and human hepatocytes. I used several in vitro and ex vivo experimental set-ups including fluorescence and radioactive labelling of lipids, to study changes in FA- uptake, de novo synthesis, storage, oxidation, and secretion. In addition, I performed fluorescence-based assays to focus on regulations affecting mitochondrial function, cell viability, proliferation and associated replication stress, and DNA damage. By using CRISPR-Cas based knock out cell models and si-RNA mediated knock downs in hepatocytes, as well as treatment of cells with different inhibitors, antagonists, and agonists, I studied involved inflammatory and metabolic pathways. To disentangle molecular mechanisms induced by FAs and pro-inflammatory cytokines, I performed a multi omics approach including lipidome, transcriptome, proteome, phosphoproteome, and thermal proteome profiling. I demonstrated that inflammatory cytokines increase FA storage in a NF-ĸB dependent manner by interference with catabolic processes. Exacerbated lipid accumulation in hepatocytes subsequently promoted mitochondrial dysfunction, apoptotic cell death and compensatory proliferation. In addition, inflammatory cytokines induced an inflammatory stress response, replication stress, and DNA damage. Importantly, the exposure to inflammatory cytokines alone and in combination with FAs lead to the downregulation of genes and proteins involved in essential metabolic processes in hepatocytes, not only specific for FA metabolism. The understanding of the effect of inflammatory cytokines on metabolic dysregulation and transcriptional control of metabolic genes in hepatocytes will help to find possible treatment options for NAFLD / NASH

Comparison of 2 concentrations of levobupivacaine in postoperative patient-controlled analgesia

peer reviewedBoth groups were similar with regard to demographics, upper level of sensory blockade (T8), and visual analog scale pain scores at rest and after coughing, as well as levobupivacaine and subcutaneous rescue morphine consumption. Motor blockade in the lower limbs was very low in both groups. Arterial blood pressure was slightly lower in the 5 mg/mL group during the first 24 hours ( P = 0.052). Five patients in the 1.5 mg/mL and 7 in the 5 mg/mL group had postoperative nausea and vomiting ( P = 0.43). No other side effects were recorded, and all of the patients were satisfied

Colon cancer cell-derived 12(S)-HETE induces the retraction of cancer-associated fibroblast via MLC2, RHO/ROCK and Ca2+ signalling

Retraction of mesenchymal stromal cells supports the invasion of colorectal cancer cells (CRC) into the adjacent compartment. CRC-secreted 12(S)-HETE enhances the retraction of cancer-associated fibroblasts (CAFs) and therefore, 12(S)-HETE may enforce invasivity of CRC. Understanding the mechanisms of metastatic CRC is crucial for successful intervention. Therefore, we studied pro-invasive contributions of stromal cells in physiologically relevant three-dimensional in vitro assays consisting of CRC spheroids, CAFs, extracellular matrix and endothelial cells, as well as in reductionist models. In order to elucidate how CAFs support CRC invasion, tumour spheroid-induced CAF retraction and free intracellular Ca2+ levels were measured and pharmacological-or siRNA-based inhibition of selected signalling cascades was performed. CRC spheroids caused the retraction of CAFs, generating entry gates in the adjacent surrogate stroma. The responsible trigger factor 12(S)-HETE provoked a signal, which was transduced by PLC, IP3, free intracellular Ca2+, Ca(2+)calmodulin-kinase-II, RHO/ROCK and MYLK which led to the activation of myosin light chain 2, and subsequent CAF mobility. RHO activity was observed downstream as well as upstream of Ca2+ release. Thus, Ca2+ signalling served as central signal amplifier. Treatment with the FDA-approved drugs carbamazepine, cinnarizine, nifedipine and bepridil HCl, which reportedly interfere with cellular calcium availability, inhibited CAF-retraction. The elucidation of signalling pathways and identification of approved inhibitory drugs warrant development of intervention strategies targeting tumour-stroma interaction

Stromal fibroblasts shape the myeloid phenotype in normal colon and colorectal cancer and induce CD163 and CCL2 expression in macrophages

Colorectal cancer (CRC) accounts for about 10% of cancer deaths worldwide. Colon carcinogenesis is critically influenced by the tumor microenvironment. Cancer associated fibroblasts (CAFs) and tumor associated macrophages (TAMs) represent the major components of the tumor microenvironment. TAMs promote tumor progression, angiogenesis and tissue remodeling. However, the impact of the molecular crosstalk of tumor cells (TCs) with CAFs and macrophages on monocyte recruitment and their phenotypic conversion is not known in detail so far. In a 3D human organotypic CRC model, we show that CAFs and normal colonic fibroblasts are critically involved in monocyte recruitment and for the establishment of a macrophage phenotype, characterized by high CD163 expression. This is in line with the steady recruitment and differentiation of monocytes to immunosuppressive macrophages in the normal colon. Cytokine profiling revealed that CAFs produce M-CSF, and IL6, IL8, HGF and CCL2 secretion was specifically induced by CAFs in co-cultures with macrophages. Moreover, macrophage/CAF/TCs co-cultures increased TC invasion. We demonstrate that CAFs and macrophages are the major producers of CCL2 and, upon co-culture, increase their CCL2 production twofold and 40-fold, respectively. CAFs and macrophages expressing high CCL2 were also found in vivo in CRC, strongly supporting our findings. CCL2, CCR2, CSF1R and CD163 expression in macrophages was dependent on active MCSFR signaling as shown by M-CSFR inhibition. These results indicate that colon fibroblasts and not TCs are the major cellular component, recruiting and dictating the fate of infiltrated monocytes towards a specific macrophage population, characterized by high CD163 expression and CCL2 production

Iron Behaving Badly: Inappropriate Iron Chelation as a Major Contributor to the Aetiology of Vascular and Other Progressive Inflammatory and Degenerative Diseases

The production of peroxide and superoxide is an inevitable consequence of aerobic metabolism, and while these particular "reactive oxygen species" (ROSs) can exhibit a number of biological effects, they are not of themselves excessively reactive and thus they are not especially damaging at physiological concentrations. However, their reactions with poorly liganded iron species can lead to the catalytic production of the very reactive and dangerous hydroxyl radical, which is exceptionally damaging, and a major cause of chronic inflammation. We review the considerable and wide-ranging evidence for the involvement of this combination of (su)peroxide and poorly liganded iron in a large number of physiological and indeed pathological processes and inflammatory disorders, especially those involving the progressive degradation of cellular and organismal performance. These diseases share a great many similarities and thus might be considered to have a common cause (i.e. iron-catalysed free radical and especially hydroxyl radical generation). The studies reviewed include those focused on a series of cardiovascular, metabolic and neurological diseases, where iron can be found at the sites of plaques and lesions, as well as studies showing the significance of iron to aging and longevity. The effective chelation of iron by natural or synthetic ligands is thus of major physiological (and potentially therapeutic) importance. As systems properties, we need to recognise that physiological observables have multiple molecular causes, and studying them in isolation leads to inconsistent patterns of apparent causality when it is the simultaneous combination of multiple factors that is responsible. This explains, for instance, the decidedly mixed effects of antioxidants that have been observed, etc...Comment: 159 pages, including 9 Figs and 2184 reference

Comparison of different concentrations of levobupivacaine for postoperative epidural analgesia

the same dose of levobupivacaine provides an equal quality of analgesia in low or high volume continuous thoracic epidural infusion with reduce haemodynamic instability and nausea in the low/high concentration groups

Cortisol reactivity in boys with attention-deficit/hyperactivity disorder and disruptive behavior problems : the impact of callous unemotional traits

There is a body of literature demonstrating an association between altered hypothalamic pituitary adrenal (HPA) axis reactivity and aggressive behavior. Aggressive and disruptive behavior also is highly prevalent in children with attention deficit/hyperactivity disorder (ADHD). Findings on HPA-axis reactivity in ADHD, however, are rather inconsistent. Specific temperamental risk factors previously were associated with a specific subtype of severe disruptive behavior. These traits might also be characterized by a distinct neurobiological profile across ADHD and disruptive behavior disorders. In this study we focus on psychopathic traits, notably callous unemotional (CU) traits. The main objective of the present study was to investigate whether two groups of ADHD patients with high or low CU traits differed in cortisol reactivity. Subjects were 36 boys with ADHD and disruptive behavior symptoms aged 8 to 14 years. Salivary cortisol probes were taken before and repeatedly after an experimental standardized stress test. Patients scoring high on CU traits showed a blunted HPA axis reactivity to the experimentally induced stress. Results underscore the need to consider specific personality traits in investigating neurobiological correlates in ADHD with disruptive behavior problems

Happier with less? Members of European environmental grassroots initiatives reconcile lower carbon footprints with higher life satisfaction and income increases

Scientists and policymakers recognize the need to address consumption and lifestyles in order to reconcile environmental and development agendas. Sustainability-oriented grassroots initiatives emerge bottom-up to create opportunities for sustainable lifestyles; yet no prior assessment has ascertained the efficacy of their members to reduce carbon footprints (CF) and enhance well-being. We compare the CF of non-members and members of grassroots initiatives in the domains of food, clothing, housing and transport. We further compare the groups by testing the influence of socio-economic variables that are typically associated with both footprint and well-being. Here we show that grassroots initiative members have 16% lower total carbon footprint, and 43% and 86% lower carbon footprints for food and clothing respectively, compared to their “non-member” regional socio-demographic counterparts. We find a higher adoption of some energy-saving behaviors for initiative members such as greater active travel distance and lower indoor temperatures in the winter, yet no significant differences in the CF of housing and transport. Interestingly, increases in income are not associated with increases in the total CF of members, while the influence of income is confirmed for the CF of the total sample. Instead, factors such as age, household size, and gender better explain the variation in the domain-specific CFs of initiative members. Finally, members show higher life satisfaction compared to non-members and are 11–13% more likely to evaluate their life positively. Our results suggest that initiative members uncover lifestyle features that not only enable lower emissions, but also reconcile emissions with income and well-being