The use of CFD modelling to optimise measurement of overflow rates in a downstream-controlled dual-overflow structure.

The measurement of the flow through complex combined sewer overflow structures in the frame of automated monitoring remains difficult. In this paper, a methodology based on the use of computational fluid dynamics (CFD) modelling in order to improve the instrumentation of a downstream-controlled dual-overflow structure is presented. The dual-overflow structure is composed of two combined sewer overflows (CSOs) connected by a rectangular channel and controlled by a downstream gate located at the entry of the Meyzieu waste water treatment plant (close to Lyon, France).

Gridifying phylogeny and medical applications on the volunteer computing platform XtremWeb-CH.

XtremWeb-CH (XWCH) is a volunteer computing middleware that makes it easy for scientists and industrials to deploy and execute their parallel and distributed applications on a public-resource computing infrastructure. XWCH supports various high performance applications, including those having large storage and communication requirements. Two high performance applications were ported and deployed on an XWCH platform.

Involvement of the Src kinase Lyn in phospholipase C-gamma 2 phosphorylation and phosphatidylinositol 3-kinase activation in Epo signalling.

We examined the role of the Src kinase Lyn in phospholipase C-gamma 2 (PLC-gamma 2) and phosphatidylinositol (PI) 3-kinase activation in erythropoietin (Epo)-stimulated FDC-P1 cells transfected with a wild type (WT) Epo-receptor (Epo-R). We showed that two inhibitors of Src kinases, PP1 and PP2, abolish both PLC-gamma 2 tyrosine phosphorylation and PI 3-kinase activity in WT Epo-R FDC-P1 cells. We also demonstrated that Epo-phosphorylated Lyn is associated with tyrosine phosphorylated PLC-gamma 2 and PI 3-kinase in WT Epo-R FDC-P1-stimulated cells.

Phosphatidylinositol 3-kinase regulates glycosylphosphatidylinositol hydrolysis through PLC-gamma(2) activation in erythropoietin-stimulated cells.

Erythropoietin (Epo)-induced glycosylphosphatidylinositol (GPI) hydrolysis was previously described to be correlated with phospholipase C-gamma 2 (PLC-gamma2) activation. Here, we analyzed the involvement of phosphatidylinositol (PtdIns) 3-kinase in GPI hydrolysis through PLC-gamma2 tyrosine phosphorylation in response to Epo in FDC-P1 cells transfected with a wild type (WT) erythropoietin-receptor (Epo-R). We showed that phosphatidylinositol 3-kinase (PtdIns 3-kinase) inhibitor LY294002 inhibits Epo-induced hydrolysis of endogenous GPI and Epo-induced PLC-gamma2 tyrosine phosphorylation in a dose-dependent manner.

Erythropoietin induction of tissue inhibitors of metalloproteinase-1 expression and secretion is mediated by mitogen-activated protein kinase and phosphatidylinositol 3-kinase pathways.

In the present study, we demonstrate that erythropoietin (Epo) induces the expression and the release of tissue inhibitors of metalloproteinase-1 (TIMP-1) in a time- and dose-dependent manner in Epo-dependent cell line UT-7 cells and in normal human erythroid progenitor cells from cord blood (CD36+) and required de novo protein synthesis. TIMP-1 was not expressed in the absence of Epo. Inhibition of the mitogen-activated protein kinase pathway by the specific inhibitors PD98059 and U0126 and of phosphatidylinositol 3-kinase by LY294002, strongly inhibited Epo-induced TIMP-1 expression and secretion.

Involvement of the p38 mitogen-activated protein kinase pathway in tissue inhibitor of metalloproteinases-1-induced erythroid differentiation.

We examined the role of the mitogen-activated protein (MAP) kinase pathway in tissue inhibitor of metalloproteinases-1 (TIMP-1)-mediated cellular effects in a human erythroleukemic cell line UT-7. We show that TIMP-1 induced both UT-7 cell erythroid differentiation and proliferation and tyrosine phosphorylation of many intracellular proteins. Using a panel of phosphospecific antibodies, we also demonstrate that phosphorylation of the p38 and c-Jun N-terminal kinases is increased by TIMP-1 whereas phosphorylation of extracellular signal-regulated kinase 1/2 is not induced.

Erythropoietin induces glycosylphosphatidylinositol hydrolysis. Possible involvement of phospholipase c-gamma(2).

We showed that erythropoietin induced rapid glycosylphosphatidylinositol (GPI) hydrolysis and tyrosine phosphorylation of phospholipase C (PLC)-gamma(2) in FDC-P1 cells transfected with the wild-type erythropoietin-receptor. Erythropoietin-induced tyrosine phosphorylation of PLC-gamma(2) was time- and dose-dependent. By using FDC-P1 cells transfected with an erythropoietin receptor devoid of tyrosine residues, we showed that both effects required the tyrosine residues of intracellular domain on the erythropoietin receptor.

Evidence of the role of protein kinase C during aclacinomycin induction of erythroid differentiation in K562 cells.

At subtoxic concentrations, aclacinomycin is effective in controlling erythroid differentiation of K562, a human erythroleukemic cell line. To better understand early events implicated in this process, we have used bisindolylmaleimide (GF109203X), an inhibitor with a high selectivity for protein kinase C (PKC). Our data show that GF109203X inhibits aclacinomycin effects on K562, evidenced by a strong reduction of hemoglobinized cells and a marked decrease of mRNA rates of erythroid genes.

Erythropoietin induces the tyrosine phosphorylation of insulin receptor substrate-2. An alternate pathway for erythropoietin-induced phosphatidylinositol 3-kinase activation.

In this report, we demonstrate that insulin receptor substrate-2 (IRS-2) is phosphorylated on tyrosine following treatment of UT-7 cells with erythropoietin. We have investigated the expression of IRS-1 and IRS-2 in several cell lines with erythroid and/or megakaryocytic features, and we observed that IRS-2 was expressed in all cell lines tested. In contrast, we did not detect the expression of IRS-1 in these cells.

Activation of Raf-1 and mitogen-activated protein kinases by erythropoietin and inositolphosphate-glycan in normal erythroid progenitor cells: involvement of protein kinase C.

In this work, we show that erythropoietin and inositolphosphate-glycan activate Raf-1 and the mitogen-activated protein kinases (MAP kinases) in normal erythropoietin-responsive cells. Using a protein kinase C (PKC) activator such as the phorbol ester, 12-O-tetradecanoyl-phorbol-13-acetate and the PKC inhibitor GF109203X, we investigated a possible involvement of PKC during activation of Raf-1 and MAP kinase by erythropoietin or inositolphosphate-glycan. We found that erythropoietin increased MAP kinase level with a maximum stimulation reached at 5-10 min.

Erythropoietin stimulates glycosylphosphatidylinositol hydrolysis in rat erythroid progenitor cells and inositolphosphate glycan modulates their proliferation.

The involvement of a glycosylphosphatidylinositol/inositolphosphate glycan (GPI/IPG) system in the erythropoietin (Epo) signal transduction was investigated. Endogenous GPI was evidenced in extracts of normal Epo-responsive cells after incorporation of [3H]glucosamine, [3H]inositol and [32P]orthophosphate. Incubation of these cells with Epo produced a rapid and transient hydrolysis of GPI with parallel release of IPG.

Glucose uptake by rat erythroid cells: the effects of erythropoietin and dexamethasone.

Erythropoietin (Ep) stimulated glucose uptake by erythroid progenitor cells in the liver of fetal rats, as measured by [3H]2-deoxy-D-glucose uptake. This dose-dependent stimulation was maximal at 0.2 U/ml Ep and decreased during cell differentiation.

Erythropoietin induces the tyrosine phosphorylation of its own receptor in human erythropoietin-responsive cells.

Using the human erythropoietin-responsive hematopoietic cell line UT-7, we showed that erythropoietin (Epo) rapidly and specifically induced the tyrosine phosphorylation of its own receptor (M(r) 75,000) and increased the tyrosine phosphorylation of other proteins of M(r) 140,000, 120,000, 95,000, 60,000, 57,000, and 42,000. Neither granulocyte-macrophage colony-stimulating factor, interleukin 3, interleukin 6, nor the kit ligand induced the phosphorylation of the M(r) 75,000 receptor protein, although these growth factors induced the phosphorylation of other proteins. Cross-linking experiments using 125I-Epo indicated that the UT-7 cells expressed three Epo receptor subunits, of M(r) 100,000, 85,000, and 75,000, among which only the M(r) 75,000 subunit was tyrosine-phosphorylated following activation with Epo.

Effects of pertussis toxin on the erythropoietin-stimulated proliferation and differentiation of erythroid-responsive cells.

Erythropoietin (Ep) stimulates the proliferation and differentiation of erythroid progenitor cells. We have investigated the possibility that guanyl nucleotide regulatory (G) proteins may be involved in Ep signal transduction. Pertussis toxin (PT) was found to inhibit Ep-stimulated [3H]-thymidine incorporation and large erythroid colony formation in a dose-dependent fashion, but had no effect on small colony formation or on Ep-induced differentiation.

In vivo effects of glucocorticoid excess on the contents of the rat fetal liver in erythroid progenitors.

Following a laparotomy of the pregnant rat at 12 days of gestation, erythroid cell suspensions prepared from the fetal livers at 14 days contained an increased proportion of progenitor cells forming colonies after 2 or 7 days of culture. When laparotomy was performed at 14 days and the fetal livers were sampled at 16 days, the opposite effects were observed. Injection of 0.

The erythropoietin receptor of rat erythroid progenitor lens. Characterization and affinity cross-linkage.

Commercially available 125I-labeled erythropoietin, obtained by genetic engineering from a human gene, was used to characterize receptors for this hormone on the cell surface of rat erythroid progenitor cells. A low number of high affinity binding sites (487 +/- 32 sites/cell, Kd = 167 +/- 14 pm) were found. Nonerythroid cells and erythrocytes did not exhibit specific binding.

Murine erythroleukaemia cells (Friend cells) possess high-affinity binding sites for erythropoietin.

Murine erythroleukaemia cells represent erythroid precursors blocked near the CFU-E or proerythroblast stage. In contrast to their non-leukaemic equivalents, neither their proliferation nor their differentiation seems to be affected by erythropoietin. However, we show in this paper that both uncommitted and committed, benzidine-positive, cells bind iodinated erythropoietin.

Induction by hemin of proliferation and of differentiation of progenitor erythroid cells responsible for erythropoietin.

Erythroid progenitor cells were obtained from rat fetal liver by immunolysis of the whole erythroid population with an antiserum directed against adult rat erythrocytes, followed by separation on a density gradient. Immediately after their isolation, these cells contained only minute amounts of globin mRNAs and their heme synthesis was negligible. In the absence of erythropoietin (Epo), they did not proliferate or differentiate.

Mode of action of erythropoietin and glucocorticoids on the hepatic erythroid precursor cells: role of prostaglandins.

The hypothesis that prostaglandins, and especially PGE2, are the second messengers of erythropoietin (Ep) and that glucocorticoids inhibit Ep action by inhibiting PG synthesis was tested on the erythroid cell line from fetal rat liver. The optimal (10(-9) M) stimulatory concentration of PGE2 did not reproduce, by far, the maximal effect of Ep on the growth of CFUE erythroid colonies. Ep did not increase PGE2 release in liquid culture media of cell suspensions made of the whole erythroid line or enriched (over 85%) in precursor cells.

Effect of the antiglucocorticoid agent RU 38486 on the dexamethasone inhibition of Friend cell differentiation.

Glucocorticoid hormones are known to inhibit the erythroid differentiation of Friend cells. The mechanism of action of these hormones has been questioned, and results suggesting an action not involving the nuclear binding of the receptors have been published. We have used the antiglucocorticoid RU 38486 to block the inhibitory effect of dexamethasone on the induced differentiation of Friend cells.

Kinetics of dexamethasone binding to the glucocorticoid receptor of intact erythroid cells from rat fetal liver.

The erythroid cells from the rat fetal liver have been shown to possess a receptor for glucocorticoids. In the present work, the characteristics of [3H]dexamethasone binding have been studied on intact cells, in order to minimize receptor degradation, and at 4 degrees C, in order to prevent the activation of the hormone-receptor complex. Dissociation kinetics were those of a first-order reaction and the value of the rate constant of dissociation was similar to the values available in the literature.

In vitro and in vivo regulation of hepatic erythropoiesis by erythropoietin and glucocorticoids in the rat fetus.

Glucocorticoids affect evolution of rat fetal liver erythropoietic tissue, where their receptors have been characterized. This paper describes erythropoietin (Ep) and dexamethasone (Dex) effects on the number of CFUE and BFUE grown from erythroid cells isolated from 14 day fetal livers. CFUE number showed a linear log dose-response towards Ep.

Binding of glucocorticosteroids to hepatic erythropoietic cells of the rat fetus.

A role for glucocorticosteroids in the evolution of the fetal liver erythron in vivo has been proposed. If direct, such an intervention implies the presence of receptor sites in the cells of the erythroid cell lines. To test this hypothesis cell suspensions were prepared from liver haematopoietic tissue obtained from rat fetuses aged 13, 14, 17 and 20 days.