Nuclear Factor-κB-Independent Anti-Inflammatory Action of Salicylate in Human Endothelial Cells

In contrast to aspirin, salicylate, its active metabolite, possesses profound anti-inflammatory properties without blocking cyclooxygenase. Inhibition of the transcription factor nuclear factor-κB (NF-κB) has been discussed to play a role in the anti-inflammatory profile of salicylate. However, NF-κB-independent effects of salicylate have been assumed but have up to now been poorly investigated. Therefore, the aim of the present study was to investigate NF-κB-independent anti-inflammatory mechanisms of salicylate in human umbilical vein endothelial cells using interleukin-4 (IL-4) as NF-κB-independent proinflammatory stimulus and P-selectin as inflammatory read-out parameter. Using quantitative real-time reverse transcriptionpolymerase chain reaction, we found that salicylate decreases IL-4-induced P-selectin expression. As judged by Western blot analysis, salicylate increased endothelial heme oxygenase-1 (HO-1) protein levels. Using both the HO-1 inhibitor tin(II) protoporphyrin IX and HO-1 antisense oligonucleotides, we causally linked the induction of HO-1 to the decrease of P-selectin. Moreover, we were interested in the signaling mechanisms leading to the up-regulation of HO-1 by salicylate. c-Jun NH2-terminal kinase (JNK) was found to be activated by salicylate, and we could causally link this activation to the induction of HO-1 by using the JNK inhibitor 1,9-pyrazoloanthrone. By applying activator protein-1 (AP-1) decoys, it was shown that the transcription factor AP-1 is crucially involved in the up-regulation of HO-1 downstream of JNK. In summary, our study introduces HO-1 as novel NF-κB-independent anti-inflammatory target of salicylate in human endothelial cells. Moreover, we elucidated the JNK/AP-1 pathway as crucial for the induction of HO-1 by salicylate

Trisubstituted pyrazolopyrimidines as novel angiogenesis inhibitors.

Current inhibitors of angiogenesis comprise either therapeutic antibodies (e.g. bevacicumab binding to VEGF-A) or small molecular inhibitors of receptor tyrosin kinases like e.g. sunitinib, which inhibits PDGFR and VEGFR. We have recently identified cyclin-dependent kinase 5 (Cdk5) as novel alternative and pharmacologically accessible target in the context of angiogenesis. In the present work we demonstrate that trisubstituted pyrazolo[4,3-d]pyrimidines constitute a novel class of compounds which potently inhibit angiogenesis. All seven tested compounds inhibited endothelial cell proliferation with IC(50) values between 1 and 18 µM. Interestingly, this seems not to be due to cytotoxicity, since none of them showed acute cytotoxic effects on endothelial cells at a concentration of 10 µM,. The three most potent compounds (LGR1404, LGR1406 and LGR1407) also inhibited cell migration (by 27, 51 and 31%, resp.), chemotaxis (by 50, 70 and 60% in accumulative distance, resp.), and tube formation (by 25, 60 and 30% of total tube length, resp.) at the non-toxic concentration of 10 µM. Furthermore, angiogenesis was reduced in vivo in the CAM assay by these three compounds. A kinase selectivity profiling revealed that the compounds prevalently inhibit Cdk2, Cdk5 and Cdk9. The phenotype of the migrating cells (reduced formation of lamellipodia, loss of Rac-1 translocation to the membrane) resembles the previously described effects of silencing of Cdk5 in endothelial cells. We conclude that especially LGR1406 and LGR1407 are highly attractive anti-angiogenic compounds, whose effects seem to largely depend on their Cdk5 inhibiting properties

Metalloporphyrins inactivate caspase-3 and -8

Activation of caspases represents one of the earliest biochemical indicators for apoptotic cell death. Therefore, measurement of caspase activity is a widely used and generally accepted method to determine apoptosis in a wide range of in vivo and in vitro settings. Numerous publications characterize the role of the heme-catabolizing enzyme heme oxygenase-1 (HO-1) in regulating apoptotic processes. Different metalloporphyrins representing inducers and inhibitors of this enzyme are often used, followed by assessment of apoptotic cell death. In the present work, we found that caspase-3-like activity, as well as activity of caspase-8 measured in either Fas (CD95) ligand-treated Jurkat T-lymphocytes or by the use of recombinant caspase-3 or -8, was inhibited by different metalloporphyrins (cobalt(III) protoporphyrin IX, tin and zinc II) protoporphyrin-IX). Moreover, employing the mouse model of Fas-induced liver apoptosis these properties of porphyrins could also be demonstrated in vivo. The metalloporphyrins were shown to inhibit caspase-3-mediated PARP cleavage. Molecular modeling studies demonstrated that porphyrins can occupy the active site of caspase-3 in an energetically favorable manner and in a binding mode similar to that of known inhibitors. The data shown here introduce metalloporphyrins as direct inhibitors of caspase activity. This finding points to the need for careful employment of metalloporphyrins as modulators of HO-1

Enzymfreisetzung und Aktivierung der Kallikrein-Kinin-Systeme bei experimenteller Pankreatitis

Das klinische Bild der akuten Pankreatitis wird entscheidend durch die sekundäre Schädigung von Herz-Kreislauf-System, Lunge und Niere bestimmt. Ziel der vorliegenden Untersuchung war es, durch Messungen in venösem Pankreasblut, Pankreaslymphe und Peritonealexsudat die Kompartimente zu bestimmen, über die die systemischen Schädigungen vermittelt werden. An anästhesierten Schweinen wurden die systemischen, hämodynamischen Parameter durch gesteuerte Volumentherapie konstant gehalten. Die Schweine wurden randomisiert der Kontrollgruppe (n = 9) oder einer der Pankreatitisgruppen zugeteilt (jeweils n = 10). Die Pankreatitis wurde durch Infusion von freier Fettsäure in die Pankreasarterien (FFS) oder durch Infusion einer 5%igen Natrium-Taurocholat-Lösung retrograd in den Pankreasgang (NaT) ausgelöst. Nach Isolation des Pankreas wurde venöses Pankreasblut, Pankreaslymphe und Peritonealexsudat gewonnen und die Aktivität von Lipase, Phospholipase A und Plasmaprokallikrein sowie die Konzentration von Organkallikrein und Kininogen bestimmt. In beiden Pankreatitismodellen fand sich ein Anstieg der Enzymaktivitäten. Die höchsten Aktivitäten fanden sich im Peritonealexsudat (Phospholipase A nach 40 min: Kontrolle 10,0 U/1, NaT 72,2 U/1). In beiden Pankreatitismodellen fanden sich außerdem Hinweise für eine Aktivierung des Organkallikrein-Kinin-Systems durch den Anstieg der Organkallikreinkonzentration und den Abfall der Gesamtkininogenkonzentration. Die stärksten Veränderungen fanden sich wieder im Peritonealexsudat (Organkallikrein nach 40 min: Kontrolle 14,7 ng/ml, NaT 452 ng/ml).The clinical course of acute pancreatitis is strongly influenced by secondary cardiac, pulmonary and renal damage. The aim of the present study was to gather information about the compartment promoting the systemic damage. Therefore the activity of lipase, phospholipase A and plasmaprokallikrein and the concentration of tissue kallikrein and kininogen were measured in portal venous blood, pancreatic lymph and peritoneal exudate. Anaesthetized pigs were subjected to fluid resuscitation to keep systemic haemodynamic parameters constant. The pancreas was isolated in situ. The pigs were randomly assigned to a control group (n = 9) or one of the two pancreatitis groups (n = 10 each). Pancreatitis was induced by i.a. infusion of free fatty acid (FFS) or retrograde infusion of 5 % sodium taurocholate intraductally (NaT). In both pancreatitis groups the activity of lipase and phospholipase A increased. The most pronounced changes were seen in the peritoneal exsudate (phospholipase A activity 40 min after induction: control 10.0 U/1, NaT 72.2 U/1). In both pancreatitis groups there was evidence for activation of the tissue kallikreinkinin system in the form of an increase in the kallikrein concentration and a decrease in the kininogen concentration. Again the changes were most pronounced in the peritoneal exsudate (tissue kallikrein 40 min after induction: control 14.7 ng/ml, NaT 452 ng/ml)

Exact Solution of an Evolutionary Model without Ageing

We introduce an age-structured asexual population model containing all the relevant features of evolutionary ageing theories. Beneficial as well as deleterious mutations, heredity and arbitrary fecundity are present and managed by natural selection. An exact solution without ageing is found. We show that fertility is associated with generalized forms of the Fibonacci sequence, while mutations and natural selection are merged into an integral equation which is solved by Fourier series. Average survival probabilities and Malthusian growth exponents are calculated indicating that the system may exhibit mutational meltdown. The relevance of the model in the context of fissile reproduction groups as many protozoa and coelenterates is discussed.Comment: LaTeX file, 15 pages, 2 ps figures, to appear in Phys. Rev.

Cdk5 controls lymphatic vessel development and function by phosphorylation of Foxc2.

The lymphatic system maintains tissue fluid balance, and dysfunction of lymphatic vessels and valves causes human lymphedema syndromes. Yet, our knowledge of the molecular mechanisms underlying lymphatic vessel development is still limited. Here, we show that cyclin-dependent kinase 5 (Cdk5) is an essential regulator of lymphatic vessel development. Endothelial-specific Cdk5 knockdown causes congenital lymphatic dysfunction and lymphedema due to defective lymphatic vessel patterning and valve formation. We identify the transcription factor Foxc2 as a key substrate of Cdk5 in the lymphatic vasculature, mechanistically linking Cdk5 to lymphatic development and valve morphogenesis. Collectively, our findings show that Cdk5-Foxc2 interaction represents a critical regulator of lymphatic vessel development and the transcriptional network underlying lymphatic vascular remodeling

Self-Organized States in Cellular Automata: Exact Solution

The spatial structure, fluctuations as well as all state probabilities of self-organized (steady) states of cellular automata can be found (almost) exactly and {\em explicitly} from their Markovian dynamics. The method is shown on an example of a natural sand pile model with a gradient threshold.Comment: 4 pages (REVTeX), incl. 2 figures (PostScript

Fuzzy Fibers: Uncertainty in dMRI Tractography

Fiber tracking based on diffusion weighted Magnetic Resonance Imaging (dMRI) allows for noninvasive reconstruction of fiber bundles in the human brain. In this chapter, we discuss sources of error and uncertainty in this technique, and review strategies that afford a more reliable interpretation of the results. This includes methods for computing and rendering probabilistic tractograms, which estimate precision in the face of measurement noise and artifacts. However, we also address aspects that have received less attention so far, such as model selection, partial voluming, and the impact of parameters, both in preprocessing and in fiber tracking itself. We conclude by giving impulses for future research

Epileptic networks in focal cortical dysplasia revealed using electroencephalography-functional magnetic resonance imaging.

Surgical treatment of focal epilepsy in patients with focal cortical dysplasia (FCD) is most successful if all epileptogenic tissue is resected. This may not be evident on structural magnetic resonance imaging (MRI), so intracranial electroencephalography (icEEG) is needed to delineate the seizure onset zone (SOZ). EEG-functional MRI (fMRI) can reveal interictal discharge (IED)-related hemodynamic changes in the irritative zone (IZ). We assessed the value of EEG-fMRI in patients with FCD-associated focal epilepsy by examining the relationship between IED-related hemodynamic changes, icEEG findings, and postoperative outcome

Forest Cover Changes in Tropical South and Central America from 1990 to 2005 and Related Carbon Emissions and Removals.

This paper outlines the methods and results for monitoring forest change and resulting carbon emissions for the 1990-2000 and 200-2005 periods carried out over tropical Central and South America. To produce our forest change estimates we used a systematic sample of medium resolution satellite data processed to forest change maps covering 1230 sites of 20 km by 20 km, each located at the degree confluence. Biomass data were spatially associated to each individual sample site so that annual carbon emissions could be estimated. For our study area we estimate that forest cover in the study area had fallen from 763 Mha (s.e. 10 Mha) in 1990 to 715 Mha (s.e. 10 Mha) in 2005. During the same period other wooded land (i.e., non-forest woody vegetation) had fallen from 191 Mha (s.e. 5.5 Mha) to 184 Mha (s.e. 5.5 Mha). This equates to an annual gross loss of 3.74 Mha·y−1 of forests (0.50% annually) between 1990 and 2000, rising to 4.40 Mha·y−1 in the early 2000s (0.61% annually), with Brazil accounting for 69% of the total losses. The annual carbon emissions from the combined loss of forests and other wooded land were calculated to be 482 MtC·y−1 (s.e. 29 MtC·y−1) for the 1990s, and 583 MtC·y−1 (s.e. 48 MtC·y−1) for the 2000 to 2005 period. Our maximum estimate of sinks from forest regrowth in tropical South America is 92 MtC·y−1. These estimates of gross emissions correspond well with the national estimates reported by Brazil, however, they are less than half of those reported in a recent study based on the FAO country statistics, highlighting the need for continued research in this area