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Testing and typing of eicosanoid-patterns

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Eicosanoids are pleiotrope mediators with essential function in most biological processes. The network of inter- and intracellular signalling requires coordinated cellular information processing. The cross-talk is characterised by complex non-linear responses to combinations of different stimuli and cells, but little is known about the density of these interactions. Here I have analysed eicosanoid interactions carried out by functional eicosanoid testing and typing (FET) in leucocytes from healthy subjects and patients suffering from inflammatory diseases. The known eicosanoid pattern scoring was extended to metabolically linked prostaglandin E2 and peptido-leukotrienes pathways, both alone and in all pair wise combinations, for basal, maximal synthesis capacity, acetylsalicylic acid, and neuropeptide modification. Eicosanoids fluctuated over twenty minutes context-dependent dynamically, demanding further data integration. The integration suggested that many stimuli converge for quantitative discrimination applying a total eicosanoid pattern score (TEP). Varying cellular activities affect FET and thereby TEP. The non-additive metabolic interactions were consistent with known mechanisms of metabolic pathway cross-talk. FET-based modelling of eicosanoid circuits most suitably reflects the fundamental impact of eicosanoids in maintaining cellular integrity of organ and body function. This might improve our present understanding of complex cellular eicosanoid interactions of inflammatory diseases and might be applied for diagnostic considerations.
Polish experience in molecular pancreatology mostly involves experimental work on intracellular signal transduction mechanisms in pancreatic acinar cells. It was found that stimulation with cholecystokinin (CCK) or exposure of pancreatic acini to reactive oxygen species induces three separate signaling cascades leading to activation of ERKs, JNK/SAPKs and p38 MAPK. In pancreatic acini, ERK cascade is also activated by epidermal growth factor (EGF). However, CCK and EGF activate this cascade by different mechanisms. EGF activates the cascade in a classical Ras-dependent manner, while CCK-induced activation of the ERK cascade is Ras-independent. Furthermore, stimulation with CCK leads to a rapid activation of PKC, which in turn may directly activate Raf family of kinases. Freshly isolated pancreatic acini contain pancreatic stellate cells which respond to EGF by activation of ERK cascade. It is possible that stimulation with CCK and EGF induces a cross-talk between acinar and stellate cells. Isolated pancreatic acinar cells irradiated with UV-B die predominantly by apoptosis while necrosis predominates among the cells subjected to supraphysiological concentrations of CCK. In pancreatic acini subjected to stressful stimuli the regulation of apoptosis may involve interaction between ERK and p38 MAPK signaling pathways. Acute pancreatitis in rats and in humans is associated with a marked increase in the plasma level of leptin which is caused by increased production of this peptide in the inflamed pancreas. It is possible that exogenous leptin protects the pancreas against development of acute pancreatitis by the activation of nitric oxide pathway.
Listeria monocytogenes, a food-borne intracellular animal and human pathogen, interacts with infected host cells both prior to entry and during the intracellular phase of infection. This review is focused on the role of secreted proteins including listeriolysin O and two distinct phospholipases C, in modulating the signal transduction of infected cells.
The ability of dihydrosphingosine to release Ca2+ from intracellular stores in neurones was investigated by combining the whole cell patch clamp technique with intracellular flash photolysis of caged, N-(2-nitrobenzyl)dihydrosphingosine. The caged dihydrosphingosine (100 μM) was applied to the intracellular environment via the CsCl-based patch pipette solution which also contained 0.3% dimethylformamide and 2 μM dithiothreitol. Cultured dorsal root ganglion neurones from neonatal rats were voltage clamped at -90 mV and inward whole cell Ca2+-activated currents were recorded in response to intracellular photorelease of dihydrosphingosine. Intracellular photorelease of dihydrosphingosine (about 5 μM) was achieved using a Xenon flash lamp. Inward Ca2+-activated currents were evoked in 50 out of 57 neurones, the mean delay to current activation following photolysis was 82±13 s. The responses were variable with neurones showing transient, oscillating or sustained inward currents. High voltage-activated Ca2+ currents evoked by 100 ms voltage step commands to 0 mV were not attenuated by photorelease of dihydrosphingosine. Controls showed that alone a flash from the Xenon lamp did not activate currents, and that the unphotolysed caged dihydrosphingosine, and intracellular photolysis of 2-(2-nitrobenzylamino) propanediol also did not evoke responses. The dihydrosphingosine current had a reversal potential of -11±3 mV (n = 11), and was carried by two distinct Cl- and cation currents which were reduced by 85% and about 20% following replacement of monovalent cations with N-methyl-D-glucamine or application of the Cl- channel blocker niflumic acid (10 μM) respectively. The responses to photoreleased dihydrosphingosine were inhibited by intracellular application of 20 μM EGTA, 10 μM ryanodine or extracellular application of 10 μM dantrolene, but persisted when Ca2+ free saline was applied to the extracellular environment. Intracellular application of uncaged dihydrosphingosine evoked responses which were attenuated by photolysis of the caged Ca2+ chelator Diazo-2. Experiments also suggested that extracellular application of dihydrosphingosine can activate membrane conductances. We conclude that dihydrosphingosine directly or indirectly mobilises Ca2+ from ryanodine-sensitive intracellular stores in cultured sensory neurones.
The study was conducted to investigate the effects of fish oil and maize oil on nuclear factor kappa B (NFκB) gene expression and the downstream pathways of intracellular signaling in spleen of chickens after lipopolysaccharide (LPS) stimulation. Two hundred eighty eight chickens were assigned in a 2×2 factorial design. Factors were dietary fat type (4.5% maize oil or 4.5% fish oil) and immunological stimulation (LPS or saline). LPS increased levels of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and arachidonic acid (AA) of spleen in chickens after the second LPS stimulation on 28 d of age. Fish oil alleviated the increase of EPA and DHA in spleen of chickens after LPS stimulation at 27 d of age. Fish oil supplementation decreased prostaglandin 2 (PGE2) production and the activity of cyclooxygenase 2 (COX2) after LPS stimulation. LPS stimulation increased the activity of phospholipase C (PLC) in spleen of chickens. And fish oil inhibited activity of PLC in spleen of chickens stimulated by LPS. Meanwhile fish oil decreased the production of inositol triphosphate (IP3) in spleen of chickens stimulated by LPS. Fish oil alleviated the mRNA abundance elevation of nuclear factor kappa B (NFκB) after LPS stimulation. These results showed that fish oil down-regulated the production of IP3 and PGE2 through inhibiting the activity of PLC and COX2 in spleen of chickens, respectively. The results of NFκB gene expression suggested fish oil might alleviate immune stress at the level of transcription.
Leptin and its receptors are newly dicovered factors involved in regulation of energy homeostasis and body weight. Leptin is specifically produced and secreted by adipocytes, but its receptor isoforms have a wide tissue distribution and in many experiments it has been demonstrated that leptin also plays a regulatory role in hematopoiesis, development, reproduction and perhaps glucose metabolism. In this short review we summarise the recent discoveries surrounding leptin, its receptors and their genes, and discuss the possible web of interactions.
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