Isolation of human promoter regions by Alu repeat consensus-based polymerase chain reaction.

Knowledge of the promoter structure is critical for an understanding of the regulation of genes. We demonstrate by analysis of 405 human genes that human promoter regions are flanked by upstream Alu repeat elements, typically at a distance of 0.5-5 kb from their protein-coding areas.

IGF-1, PDGF and CD18 are adherence-responsive genes: regulation during monocyte differentiation.

Unstimulated mononuclear cells express IGF-1, PDGF-A and PDGF-B mRNA, but not a number of other genes coding for growth factors or cytokines, as we demonstrated previously. The main focus of the present investigation was to compare gene expression of mononuclear cells unstimulated in suspension with gene expression of monocytes stimulated by adherence. mRNA levels of IGF-1-A and -B, PDGF-A, -B, PD-ECGF, basic FGF, acidic FGF, TGF-alpha, TGF-beta 1, and IGF-2 were sought for and quantified with our sensitive RT-PCR method (3n-PCR).

Regulation of angiogenesis by SPARC and angiostatin: implications for tumor cell biology.

The formation of blood vessels through endothelial cell proliferation from extant vasculature (angiogenesis) is a prerequisite for the remodeling, regeneration and repair of tissue. In pathological processes, angiogenesis is a major limiting step in tumor growth and is necessary for tumor formation at metastatic sites. Angiogenesis consists of a sequence of events that include (i) dissolution of the basement membrane, (ii) migration and (iii) proliferation of endothelial cells, (iv) formation of the vascular loop, and (v) formation of a new basement membrane.

The human leukotriene A4 hydrolase gene is expressed in two alternatively spliced mRNA forms.

We identified a novel short (s-) mRNA of the human leukotriene-A4 (LTA4) hydrolase using sequential reverse transcriptase PCR mapping. The s-mRNA is generated by skipping an 83 bp exon located in the 3' coding region and suggests the expression of an LTA4 hydrolase isoform with a calculated molecular mass of 59 kDa and a distinct C-terminus. Both LTA4 hydrolase mRNAs are constitutively expressed in blood cells, endothelial cells, smooth muscle cells, fibroblasts and tumour cells.

Human mononuclear cells express 12-LX: coordinated mRNA regulation with 5-LX and FLAP genes.

Lipoxygenases (LXs) catalyze formation of leukotrienes and hydroxy-eicosatetraenoic acids (HETEs), proinflammatory, and spasmogenic autacoids that are critical for host defense systems. We studied the expression and regulation of LX genes (12-LX, 5-LX, and 15-LX) and the 5-lipoxygenase activating protein (FLAP) in human mononuclear cells (MNC) and granulocytes using a quantitative reverse transcription polymerase chain reaction (RT-PCR) technique. We show that 12-LX mRNA is constitutively expressed in resting platelet-free MNC.

Thrombosis and inflammation as multicellular processes: significance of cell-cell interactions.

Platelet activation as a result of vascular injury provokes endothelial cells to respond in a manner which limits or reverses the occlusive consequences of platelet accumulation. If the agonistic forces are strong, platelet accumulation is irreversible. In vitro data from our laboratory have repeatedly demonstrated that platelets become unresponsive to all agonists when in proximity to endothelial cells.

Growth factor mRNA profiles in unstimulated human mononuclear cells: identification of genes which are constitutively and variably expressed.

Quiescent human mononuclear cells (MNC) as studied ex vivo express highly specific mRNA patterns of growth factors: We recently demonstrated that unstimulated MNC constitutively express the genes for the A and B chains of platelet-derived growth factor (PDGF). This expression was down-regulated by dietary omega-3 fatty acids. We now report that unstimulated human MNC express the genes for platelet-derived endothelial cell growth factor (PD-ECGF), insulin-like growth factor (IGF-1A, -1B) and transforming growth factor-beta 1 (TGF-beta 1).

Dietary omega-3 fatty acids lower levels of platelet-derived growth factor mRNA in human mononuclear cells.

Platelet-derived growth factor (PDGF) is a potent mitogen thought to propagate atherosclerosis and other proliferative or inflammatory diseases. Some of these diseases are ameliorated in humans by ingestion of omega-3 fatty acids. We investigated mRNA expression of both PDGF-A and PDGF-B in quiescent peripheral blood mononuclear cells from healthy male volunteers.

n-3 fatty acids and cysteinyl-leukotriene formation in humans in vitro, ex vivo, and in vivo.

In humans, dietary n-3 fatty acids ameliorate some chronic inflammatory diseases. In contrast, however, dietary n-3 fatty acids had no effect in patients with bronchial asthma. In bronchial asthma, the cysteinyl-leukotrienes C4, D4, and E4, formed from arachidonic acid, are considered important mediators.

Molecular cloning and sequencing of a murine pgk-1 pseudogene family.

Seven genomic mouse DNA fragments carrying pgk-1-homologous regions have been cloned and sequenced. They have to be classified as processed genes because intervening sequences, present in their productive counterpart, are absent. Four pseudogenes (I-IV) represent nearly the complete sequence of pgk-1 cDNA.