1. Myosin was isolated from human right- and left-atrial and -ventricular myocardium, and examined both in adult subjects and at different stages during pre- and post-natal development. 2. The myosin light-chain subunits in the atria and ventricles were different when characterized by isoelectric focusing and subsequent two-dimensional poly-acrylamide-gel electrophoresis. 3. No differences were observed between the light-chain subunits in the right and left ventricle at any stage of development. 4. The foetal ventricle contained a characteristic light chain that was a major component throughout the latter half of gestation. This foetal light chain, which disappeared in the postnatal period, could not be distinguished from adult atrial light chain 1 on two-dimensional electrophoresis. 5. Myosin in the adult atria, particularly the left, contained components similar to ventricular light-chain components. 6. The possible stimuli for the observed changes in myosin light-chain expression are discussed in relation to the known physiological changes occurring during development.
Exposure to mineral dusts is associated with the development of chronic airflow obstruction, probably mediated in part by dust-induced fibrosis of the small airways. To investigate the mechanism of fibrosis, we exposed rat tracheal explants to amosite asbestos, iron oxide, or titanium dioxide. Explants were then maintained in air organ culture, and the expression of genes encoding for various mediators and matrix components assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). At 7 d, all dusts produced significant increases in platelet-derived growth factor-A (PDGF-A) and transforming growth factor-beta1 (TGF-beta1) gene expression compared with control; asbestos and titanium dioxide produced increases in PDGF-B, and titanium dioxide increased TGF-alpha expression. Only asbestos caused increases in procollagen expression. No dust increased expression of tumor necrosis factor-alpha (TNF-alpha), fibronectin, or tropoelastin. Elevations in these factors coincided temporally with transport of particles into the epithelium and then to the subepithelial space. By in situ hybridization, TGF-beta gene expression was found in both the epithelium and subepithelial (interstitial) space, and PDGF-B and procollagen gene expression in the subepithelial space. Chemical analysis showed a small increase in hydroxyproline, a measure of collagen content, in asbestos-treated explants. We conclude that mineral dusts can induce airway wall fibrosis by directly upregulating proliferative and fibrogenic mediators as well as matrix components in the airway epithelium and interstitium, and that neither airspace nor circulating inflammatory cells are required for these effects. Different mineral dusts produce different patterns of reaction.
Phagocytosis is defined as the ingestion of particulates over 0.5 microm in diameter and is associated with cells of the immune system such as macrophages or monocytes. Neurones are not generally recognized to be phagocytic. Using light, confocal, time-lapse and electron microscopy, we carried out a wide range of in-vitro and in-vivo experiments to examine the phagocytic capacity of different neuronal cell types. We demonstrated phagocytosis of material by neurones, including cell debris and synthetic particles up to 2.8 microm in diameter. We showed phagocytosis in different neuronal types, and demonstrated that debris can be transported from neurite extremities to cell bodies and persist within neurones. Flow cytometry analysis demonstrated the lack of certain complement receptors on neurones but the presence of others, including integrin receptors known to mediate macrophage phagocytosis, indicating that a restricted set of phagocytosis receptors may mediate this process. Neuronal phagocytosis occurs in vitro and in vivo, and we propose that this is a more widespread and significant process than previously recognized. Neuronal phagocytosis may explain certain inclusions in neurones during disease, cell-to-cell spread of disease, neuronal death during disease progression and provide a potential mechanism for therapeutic intervention through the delivery of particulate drug carriers.
Myosin light chain subunits from adult human atria and ventricle were characterized by two-dimensional gel electrophoresis. Atrial (ALC-1 and ALC-2) differed from ventricular (VLC-1 and VLC-2) light chains. Foetal (18--21 week) ventricle contained VLC-1 plus a foetal light chain (FLC-1) which could not be distinguished from adult ALC-1. Myosin FLC-1 was gradually lost from the ventricle and replaced by VLC-1 during foetal development and in the immediate postnatal period.
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