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Similarities of an autoantigen in aneurysmal disease of the human abdominal aorta to a 36-kDa microfibril-associated bovine aortic glycoprotein.

Immunoglobulin G (IgG) from human aneurysmal aorta was used to partially purify an aortic protein with an apparent MW approximately 80 kDa. Amino acid sequencing of a tryptic digest revealed two sequences with homology to mouse tenascin-X. The autoimmune IgG was then shown to react with purified human tenascin, and a rabbit polyclonal anti-human tenascin antibody was found to react with the purified autoantigen. These observations suggest that the autoantigen of abdominal aortic aneurysm disease may be homologous to a calcium-binding member of the tenascin superfamily that has been identified by others in pig and cow.

Amino Acid Sequence↗

The influence of congo red on the cell wall and (1----3)-beta-D-glucan microfibril biogenesis in Saccharomyces cerevisiae.

Congo red was applied to growing yeast cells and regenerating protoplasts in order to study its effects on wall biogenesis and cell morphogenesis. In the presence of the dye, the whole yeast cells grew and divided to form chains of connected cells showing aberrant wall structures on both sides of the septum. The wall-less protoplasts in solid medium with the dye exhibited an abnormal increase in volume, regeneration of aberrant cell walls and inability to carry out cytokinesis or protoplast reversion to cells. In liquid medium, the protoplasts synthesized glucan nets composed mainly of thin fibrils orientated at random, whereas normally, in the absence of dye, the nets consist of rather thick fibrils, 10 to 20 nm in width, assembled into broad ribbons. These fibrils are known to consist of triple 6/1 helical strands of (1----3)-beta-D-glucan aggregated laterally in crystalline packing. The thin fibrils (c. 4 to 8 nm wide) can contain only a few triple helical strands (c. 1.6 nm wide) and are supposed to be prevented from further aggregation and crystallization by complexing with Congo red on their surfaces. Some loose triple 6/1 helical strands (native elementary fibrils) are also discernible. They represent the first native (1----3)-beta-D-glucan elementary fibrils depicted by electron microscopy. The effects of Congo red on growth and the wall structure in normal cells and regenerating protoplasts in solid medium can be explained by the presence of a complex which the dye forms with (helical) chain parts of the glucan network and which results in a loss of rigidity by a blocked lateral interaction between the helices.

Cell Division↗

Spectrophotometric evaluation of the adhesion of blood platelets to collagen and microfibrils.

We present an easy method in which the adhesion of platelets to collagen or to MFs was measured in a spectrophotometer, after an incubation of hypercitrated platelet rich plasma (PRP), or of a platelet suspension, with an inducer, followed by the filtration of non adhering platelets through translucent Isopore membrane filters (pore diameter = 5 microns). The adhering platelets, which are retained on the filter, were stained by Coomassie blue to quantify the adhesion by the simple reading of the O.D. 580 nm of the stained platelets which appear as a blue spot on the translucent membranes.

Blood Platelets↗

Fibrillin secretion and microfibril assembly by Marfan dermal fibroblasts.

The Marfan syndrome has been linked to the FBN1 gene encoding the microfibrillar glycoprotein fibrillin. To date, there have been no descriptions of microfibrillar abnormalities characteristic of this connective tissue disorder, although biochemical analyses have highlighted apparent abnormalities in fibrillin synthesis, secretion and processing. We have conducted a biochemical and ultrastructural investigation of fibrillin expression and assembly by a panel of dermal fibroblast lines from patients with Marfan syndrome and related diseases. The study has highlighted marked differences between cells in terms of secretion and aggregation of newly-synthesised fibrillin. In addition, electron microscopic visualization of fibrillin assemblies has clearly demonstrated for the first time the plethora of microfibrillar abnormalities that underlie this heterogeneous disorder. These data emphasize the molecular complexity that is a feature of the diverse clinical phenotypes exhibited by Marfan patients.

Autoradiography↗