Vascular endothelial growth factor receptor KDR tyrosine kinase activity is increased by autophosphorylation of two activation loop tyrosine residues.

Vascular endothelial growth factor is an important physiological regulator of angiogenesis. The function of this endothelial cell selective growth factor is mediated by two homologous tyrosine kinase receptors, fms-like tyrosine kinase 1 (Flt-1) and kinase domain receptor (KDR). Although the functional consequence of vascular endothelial growth factor binding to the Flt-1 receptor is not fully understood, it is well established that mitogenic signaling is mediated by KDR.

Red cell folate in elderly patients with myelodysplastic syndrome.

During a 7-month period a prospective study of 71 anaemic patients (29 males and 42 females) over the age of 50 was undertaken in order to identify patients with myelodysplastic syndrome (MDS). The mean values of mean corpuscular volume (MCV), serum ferritin, folate, vitamin B12 and red cell folate (RCF) of patients grouped according to the diagnosis were compared to those observed in age-matched blood donors. Forty-four of the 71 elderly patients showed macrocytic anaemia: 21 of them had gastric disease and the remaining 23 MDS.

Simultaneous differentiation and quantitation of erythroblasts and white blood cells on a high throughput clinical haematology analyser.

After the neonatal period, the presence of nucleated red blood cells (NRBC) in peripheral blood is indicative of pathology. Despite the clinical utility of such measurements, automated NRBC counting has hitherto not been available on routine automated blood cell counting analysers. To address this, an automated method for the analysis of NRBC was developed and incorporated into the Abbott Cell Dyn 4000 (CD4000) haematology analyser.

Paracrine expression of a native soluble vascular endothelial growth factor receptor inhibits tumor growth, metastasis, and mortality rate.

Vascular endothelial growth factor (VEGF) is a potent and selective vascular endothelial cell mitogen and angiogenic factor. VEGF expression is elevated in a wide variety of solid tumors and is thought to support their growth by enhancing tumor neovascularization. To block VEGF-dependent angiogenesis, tumor cells were transfected with cDNA encoding the native soluble FLT-1 (sFLT-1) truncated VEGF receptor which can function both by sequestering VEGF and, in a dominant negative fashion, by forming inactive heterodimers with membrane-spanning VEGF receptors.

Evaluation of the Abbott Cell Dyn 4000 automated fluorescent reticulocyte measurements: comparison with manual, FACScan and Sysmex R1000 methods.

The Abbott Cell Dyn 4000 (CD4000) is the first haematology analyser in which fully-automated reticulocyte measurements can be routinely determined by fluorescence as part of the full blood count. This communication reports the first evaluation of this method which was undertaken by three independent reference laboratories in Belgium, Germany and Italy. A total of 695 different samples was entered into the study which was designed to compare CD4000 reticulocyte information (enumeration and qualitative maturational data) with results determined in parallel with the existing manual (supravital staining) reference procedure, and two semi-automated fluorescent assays (Becton Dickinson FACScan and Sysmex R1000 instruments).

Red cell ferritin, a marker of iron deficiency in hemodialysis patients.

Estimation of red cell ferritin (RCFer) may give a good indication of iron supply to the erythron and it may therefore be clinically useful for the detection of functional iron deficiency. In a cross-sectional study of hemodialysis patients on erythropoietin (EPO) therapy and regular oral iron we have compared the RCFer levels with conventional indicators of iron status. The patients studied, 19 female, 48 male, mean age 62 +/- 3.

Risk of neural tube defect-affected pregnancy is associated with a block in maternal one-carbon metabolism at the level of N-5-methyltetrahydrofolate:homocysteine methyltransferase.

The disposition of whole blood mono-to hexaglutamyl methylfolate and plasma homocysteine (HCY) was used to evaluate potential lesion sites in one-carbon metabolism which could be responsible for neural tube defect(NTD)-affected pregnancies. An isocratic high-performance liquid chromatographic system (HPLC) with photodiode array detection was used to quantify and speciate whole-blood methylfolate into mono-, di-, tri-, tetra-, penta-, and hexaglutamate forms. This technique was also used with off-line radioassay to identify nonmethyl whole-blood folates.

Identification of a natural soluble form of the vascular endothelial growth factor receptor, FLT-1, and its heterodimerization with KDR.

A soluble form of the vascular endothelial growth factor (VEGF) receptor FLT-1 was identified in conditioned culture medium of human umbilical vein endothelial cells. The endogenous soluble FLT-1 (sFLT-1) receptor is chromatographically and immunologically similar to recombinant human sFLT-1 and binds [125I]VEGF with a comparable high affinity. Human sFLT-1 is shown to form a VEGF-stabilized complex with the extracellular domain of KDR in vitro, suggesting that not only full-length receptors are capable of forming ligand-induced heterodimeric complexes but also sFLT-1 can form a dominant negative complex with the mitogenically competent full-length KDR receptor.

Persistence of bacteria on antibiotic loaded acrylic depots. A reason for caution.

Bacterial growth on the surface of antibiotic loaded acrylic cement was examined in an in vitro model. Tobramycin or vancomycin impregnated discs were incubated in broth containing either Staphylococcus epidermidis or Staphylococcus aureus organisms. At 24, 48, and 96 hours, the broth and the surface of the acrylic discs were examined for viable organisms.

Bacterial growth on antibiotic-loaded acrylic cement. A prospective in vivo retrieval study.

Twenty-three patients with intraoperative culture-proven periprosthetic infection of the hip or knee were enrolled in a prospective cement retrieval study. All were treated with a two-stage technique using antibiotic-loaded acrylic cement as an antibiotic depot. Staphylococcus epidermidis was the most commonly isolated organism (19 of 23 cases).

Re-establishment of erythropoietin responsiveness in end-stage renal failure following renal transplantation.

Re-establishment of erythropoietin (EPO) secretion following renal transplantation is poorly understood. The development of sensitive EPO radioimmunoassay has enabled further study of this phenomenon. Forty-one adult patients were studied during the first 16 weeks following renal transplantation.

Eukaryotic methionyl aminopeptidases: two classes of cobalt-dependent enzymes.

Using partial amino acid sequence data derived from porcine methionyl aminopeptidase (MetAP; methionine aminopeptidase, peptidase M; EC 3.4.11.

Quantitative reticulocyte analysis may be of benefit in monitoring erythropoietin treatment in dialysis patients.

Reticulocyte responses to low-dose erythropoietin (EPO) were monitored using automated flow cytometric analysis. Sixteen adult dialysis patients were treated with 1,000 U of recombinant human EPO (rHuEPO), subcutaneously, thrice weekly (mean dose 15.7, SD 3.

Serum erythropoietin during autologous bone marrow transplantation: relationship to measures of erythroid activity.

Marked elevation of serum erythropoietin (sEPO) occurs following high dose chemotherapy for malignant disease. It has been proposed that the subsequent fall in sEPO constitutes a relative erythropoietin (EPO) deficiency, prompting trials of recombinant EPO to reduce red cell transfusion during chemotherapy. We have investigated these phenomena by serial estimations of reticulocytes and sEPO in 11 autologous marrow transplant recipients.

Toxicological foundations of ecological risk assessment: biomarker development and interpretation based on laboratory and wildlife species.

Ecological risk assessments based on chemical residue analysis and species demographics tend to ignore the bioavailability and bioaccumulation of the chemicals of concern. This study describes the incorporation of mechanistically based biomarkers into an ecological risk assessment of a poly-cyclic aromatic hydrocarbon (PAH)-contaminated site. A combination of soil residue analysis, tissue residue analysis, biomarkers in one-site trapped animals and biomarkers in animals confined to enclosures was used.

Temporary antibiotic loaded acrylic hip replacement: a novel method for management of the infected THA.

Postoperative infection after hip joint replacement is an uncommon but potentially devastating complication in contemporary orthopaedics. Management in two stages is the more favored approach in North America. This introduces difficulty with patient management in the interval between stages, delays rehabilitation, and introduces technical difficulty during the second stage.

The effect of cytokines on the in-vitro erythropoietin bioassay.

The effects of various concentrations of granulocyte macrophage colony stimulating factor (GM-CSF), gamma interferon (gamma IFN) and interleukins 1 alpha (IL-1 alpha), 1 beta (IL-1 beta) and 3 (IL-3) on the anaemic mouse spleen cell bioassay for erythropoietin (EPO) were investigated. Addition of IL-3 and GM-CSF at various concentrations had no effect on EPO stimulated 3H thymidine incorporation. However the addition of IL-1 alpha, IL1-beta and gamma IFN (3.

Two-stage exchange arthroplasty using a functional antibiotic-loaded spacer in the treatment of the infected knee replacement: the Vancouver experience.

Two-stage exchange arthroplasty is currently the method of choice in the treatment of the infected knee replacement. The prosthesis of antibiotic-loaded acrylic cement (PROSTALAC) is a temporary, antibiotic-loaded functional prosthesis that is used as an interim spacer in two-stage exchange arthroplasty. In this prospective series, we report on the early results of the use of the PROSTALAC knee spacer in two-stage exchange arthroplasty of infected knees.

Specificity of vascular endothelial cell growth factor receptor ligand binding domains.

Vascular endothelial cell growth factor binds with high affinity to FLT and KDR, two homologous tyrosine kinase receptors expressed on vascular endothelial cells. Placental growth factor, a vascular endothelial cell growth factor homologue, also binds with high affinity to the extracellular domains of FLT but not to the extracellular region of KDR. Vascular endothelial cell growth factor binds competitively with placental growth factor to the extracellular ligand binding domains of FLT, indicating that both ligands probably complex to overlapping or identical regions of this receptor.

Organochlorine contaminant assessment in great blue herons using traditional and nonlethal monitoring techniques.

A nonlethal method is discussed for the evaluation of contaminant concentrations in whole eggs. Concentrations of pentachlorobenzene, hexachlorobenzene, DDE, and the PCB congeners, BZ-60, BZ-118, BZ-138, BZ-180, and BZ-170 were quantified in tissue samples from great blue herons (Ardea herodias). All tissues within whole eggs from two colonies were homogenized together and analysed for these chlorinated contaminants.