Epitope-labeled soluble human interleukin-5 (IL-5) receptors. Affinity cross-link labeling, IL-5 binding, and biological activity

The human receptor for the potent eosinophilopoietic cytokine interleukin-5 (IL-5) consists of two components: a 60-kDa ligand-binding alpha chain (IL-5 alpha R) and a 130-kDa beta chain (IL-5 beta R). Three ectodomain constructs of the alpha chain (alpha RED) bearing C-terminal epitope tags were engineered and expressed in baculovirus-infected Sf9 cells. Each recombinant alpha chain was secreted into the medium, maximum expression occurring 72 h post-infection. The various soluble alpha chains were shown by affinity cross-link labeling and competition with unlabeled IL-5 to bind recombinant human (rh) 125I-IL-5 specifically with an ED50 of 2-5 nM. The epitope tag provided a simple purification of the receptor from conditioned medium using immunoaffinity chromatography. The purified material had an apparent molecular mass of 43 kDa and was heterogeneously glycosylated. Sedimentation analysis revealed a 1:1 association of the purified epitope-tagged soluble receptor with its ligand, resulting in the formation of a 70-74-kDa complex. Circular dichroism analysis revealed that the soluble alpha chain existed with a significantly ordered structure consisting of 42% beta-sheet and 6% alpha-helix. Such analyses combined with fluorescence spectrometry suggested that ligand-receptor complex formation in solution resulted in minimal conformational changes, consistent with the suggestion that the membrane-associated form of the alpha chain itself has minimal signal transduction capability. Surface plasmon resonance studies of the interaction of the purified alpha RED with immobilized rhIL-5 revealed a specific, competable interaction with a dissociation constant of 9 nM. Preincubation of an IL-5-dependent cell line with the epitope-tagged alpha RED also dose-dependently neutralized rhIL-5-induced proliferation. These data demonstrate that biologically active epitope-tagged recombinant soluble IL-5 receptors are facile to produce in large quantities and may have therapeutic utility in the modulation of IL-5-dependent eosinophilia in man.     

Antibodies to snowshoe hare virus of the California group in the snowshoe hare (Lepus americanus) population of Nova Scotia

Serological study showed the presence of antibodies to snowshoe hare virus in 6--14% of snowshoe hares (Lepus americanus) samples in Nova Scotia. This report extends the known range of this virus in North America.     

Insonation and impedance analysis in graft surveillance

Seventy consecutive patients with infrainguinal bypass grafts entered a 1-year graft surveillance programme involving colour duplex scanning, direct graft insonation and computer-assisted impedance analysis. Graft patients with a positive duplex scan, high frequencies on graft insonation or an impedance value above 0.50 subsequently underwent arteriography. Sixteen patients were excluded before the initial surveillance visit. The 54 remaining patients with grafts (30 vein, 24 synthetic) underwent a total of 137 surveillance visits, with 21 grafts confirmed to be 'at risk'. The sensitivity of an impedance value above 0.55 in identifying these grafts was 86 per cent, rising to 95 per cent when combined with graft insonation. Duplex scanning did not identify any abnormalities in 11 grafts that were either shown by arteriography to be 'at risk' or occluded before arteriography. Impedance measurement and graft insonation are simple screening techniques with a high sensitivity (when combined), which identify 'at risk' infrainguinal grafts. Positive graft insonation or an impedance value over 0.55 will identify all 'at risk' vein grafts while minimizing the number of unnecessary arteriograms.     

Glial cells in the enteric nervous system contain glial fibrillary acidic protein

The complex nervous networks found throughout the mammalian gut--the enteric nervous system--are histologically, ultrastructurally, and, to some extent, functionally--similar to the central nervous system. The glial cells of the small enteric ganglia are generally classified as Schwann or satellite cells, since they are found in the peripheral nervous system, possess nuclei which ultrastructurally resemble those of Schwann cells and are derived from the neural crest. However, it has been argued that these cells resemble astrocytes of the central nervous system with respect to gross and fine structure, and their relationship with the enteric neurones and their processes. In immunohistochemical studies of these cells, both in frozen sections of gut wall and in tissue culture preparations of the enteric plexuses, we found evidence that the enteric glial cells are rich in glial fibrillary acidic protein (GFAP), a protein associated with the 100 A glial intermediate filaments, and hitherto believed to be specific to astrocytes of the central nervous system only.     

Acute fatty liver of pregnancy

Acute fatty liver is a rare but potentially fatal complication of the third trimester of pregnancy. Significant improvements in morbidity and mortality have been reported in the last several years. Despite accumulation of more data about the disease, the exact pathogenesis is unknown. Many women are initially misdiagnosed with other more common causes of liver dysfunction during pregnancy. It is possible that acute fatty liver is an atypical form of preeclampsia because 30% to 40% of women with acute fatty liver also have preeclampsia. Supportive care and expeditious delivery represent the only known treatment. More data are needed about acute fatty liver of pregnancy, but the rare nature of the disease and the likelihood that most cases are not reported in the literature limit the ability systematically to study causation, disease process, and treatment options. Because of the serious condition of most women who develop acute fatty liver of pregnancy, collaboration between critical care and perinatal care providers is essential for optimal maternal-fetal outcomes.     

Negligible prevalence of antibodies against Trypanosoma cruzi among blood donors in the southeastern United States

The ultrastructure of alpha-bacteroids in relation to the fat-body cells of the lantern bug Pyrops candelaria was described. The fat-body-cell cytoplasm contained numerous mitochondria, rough endoplasmic reticula, vacuoles, and storage granules. Its nucleus had scattered chromatin materials. The alpha-bacteroid was enveloped by three membrane layers, namely, the plasma membrane, the cell wall, and the membrane envelope. Its cytoplasm contained amorphous dense bodies. The bacteroid reproduced by binary fission. Tracheoles were also found among fat-body cells.