Murine Ia-associated invariant chain's processing to complex oligosaccharide forms and its dissociation from the I-Ak complex

The processing of murine invariant chain (Ii) to a cell surface form bearing complex N-linked oligosaccharides has been demonstrated in the B cell lymphoma, AKTB-1b. In addition, the rate of processing of pulse-labeled Ii has been determined relative to its rate of dissociation from the alpha/beta complex of I-Ak. Ii, alpha-, and beta-chains were immunoprecipitated with anti-I-Ak or anti-Ii monoclonal antibodies. The heretofore uncharacterized complex oligosaccharide form of Ii (Ii-c) was identified in gel-purified immunoprecipitates by peptide mapping with reverse-phase HPLC. Ii-c is resistant to deglycosylation by Endo H, which is specific for high-mannose N-linkages, but can be digested with Endo F, a glycosidase capable of cleaving both complex and high-mannose N-linked oligosaccharides. Immunoprecipitation of surface iodinated cells indicates that Ii-c is expressed on the plasma membrane. Pulse-chase metabolic labeling data show that the processing of Ii to Ii-c occurs with a t1/2 of about 120 min. In contrast, the processing of both alpha- and beta-chains of I-Ak to complex forms occurs with a t1/2 of 15 to 20 min. Our data show that Ii-hm begins to dissociate rapidly from the I-Ak complex after 100 to 120 min of chase. Only a small amount (less than 5% on a per mole basis) of Ii-c was found associated with the I-Ak complexes after 300 min of continuous metabolic labeling. These results are consistent with Ii serving as a carrier for Ia antigens as they are transported to the cell surface. In addition, they suggest that the processing of Ii to Ii-c, or a late processing event of the alpha- and beta-chains, such as their sialylation, may be a possible mechanism for inducing the dissociation of Ii from the I-Ak complex.     

Capillary permeability in the pancreas and colon: restriction of exogenous and endogenous molecules by fenestrated endothelia

The permeability properties of fenestrated capillaries in the colon and exocrine and endocrine pancreas to exogenous and endogenous molecules were examined. The exogenous tracers horseradish peroxidase (Einstein-Stokes radius [ESR], 3.0 nm), hemoglobin (ESR, 3.2 nm), and ferritin (ESR, 6.1 nm) were injected intravenously and allowed to circulate for 5-90 min. Tissues were removed and processed for cytochemical or standard electron microscopic examination. The endogenous plasma proteins albumin (ESR, 3.5 nm) and IgG (ESR, 5.5 nm) were localized by immunocytochemistry using the protein A-gold technique. All vessels examined were permeable to HRP in less than 5 min. In contrast, these vessels were restrictive to the slightly larger hemoglobin molecule (60-min circulation) and to ferritin (90-min circulation). Capillaries in the exocrine and endocrine pancreas were restrictive to albumin and IgG. These results demonstrate the presence of fenestrated capillary beds, in addition to the choriocapillaris, that are restrictive to molecules with ESR greater than or equal to 3.2 nm. Capillaries in the mucosa of the colon were restrictive to hemoglobin and ferritin but did not restrict albumin or IgG. This indicates that these vessels are of the permeable type. However, the rate of transendothelial movement of molecules is slower than that of other permeable vessels, such as in the ileo-jejunum. This study has provided further evidence for the existence of fenestrated endothelia that are restrictive to exogenous and/or endogenous molecules.     

Induction of autocrine epidermal growth factor receptor ligands in human keratinocytes by insulin/insulin-like growth factor-1

Autocrine activation of the epidermal growth factor (EGF) receptor on keratinocytes has been recognized as an important growth regulatory mechanism involved in epithelial homeostasis, and, possibly, hyperproliferative diseases. Insulin-like growth factor (IGF)-1 and insulin have been shown to be paracrine keratinocyte mitogens that bind to the type I IGF receptor which is expressed on actively proliferating keratinocytes in situ. In this report, we demonstrate that IGF-1/insulin induced production of keratinocyte-derived autocrine growth factors that bind to the EGF receptor. Increased steady-state mRNA levels for transforming growth factor alpha (TGF-α) and for amphiregulin (AR) were observed upon incubation of keratinocytes with mitogenic concentrations of IGF-1. IGF-1 also induced production and secretion of TGF-α and AR proteins as detected by immunoassays. An EGF receptor antagonistic monoclonal antibody abolished the mitogenic effect of IGF-1 on cultured keratinocytes. These results suggest that stimulation of keratinocyte growth by IGF-1 requires activation of an EGF receptor-mediated autocrine loop. © 1995 Wiley-Liss, Inc.     

Formation of septa in the bulbus cordis of a turtle and a lizard

The epithelium lining the intrahepatic bile ducts of normal adult mice consists of a single layer of cuboidal or low columnar cells and has ultrastructure comparable to that described previously (Rouiller and Jézéquel, '65). Some of the epithelial cells, however, exhibit such particular features as dilatation of granular endoplasmic reticulum cisternae, polysome formation of ribosomes and the presence of active forms of the Golgi apparatus, numerous lysosome-like bodies and apical projections and blebs. Postnatal cholecystectomy does not induce any qualitative changes in the epithelial fine structure, but results in a significant increase in number of the particular structures mentioned. Therefore, the cholecystectomy is thought to stimulate the secretory activity of the epithelial cells, and such stimulation appears due to the absence of a possible activity of epithelial secretion in the gallbladder.     

Ultrastructure of Nitrosospira sp. X101 with crystalline outer membrane protein (COMP)

Abstract The outer membrane (OM) structure of Nitrosospira sp. X101 was studied by different electron microscopic techniques and SDS-PAGE. A crystalline outer membrane protein was visible in freeze-etched cells, occasionally seen also in the thin sectioned cells, but was difficult to see in a negatively-stained preparation. The lattice probably consists of large globular protein subunits with a hexagonal arrangement. The molecular weights of the major proteins in the cell envelope are 35 kDa, 40 kDa and 42 kDa.     

Mutagenic activity of nine N,N-disubstituted hydrazines in the Salmonella/mammalian microsome assay.

The mutagenic activity of N,N-dimethyl-, N,N-diethyl-, N,N-dibutyl-, N,N-diisobutyl-, N,N-di(p-tolyl)-, N-ethyl-N-phenyl-, N,N-dibenzyl-, N,N-diphenyl- and N,N-diisopropylhydrazine was examined in the Salmonella/mammalian microsome assay using the strains TA1535, TA1537, TA97, TA98, TA100, TA102 and TA1530. All nine hydrazines were mutagenic in at least one tester strain, although of borderline significance for some of the compounds. The mutagenic potencies of the hydrazines varied 2-3 orders of magnitude, from very weak to moderate mutagenic activity. In general, the addition of S9 resulted in a lowering of the mutagenic activity and a lowering of the toxic properties of the hydrazines. The test results were relatively difficult to evaluate due to toxic effects of many of the test compounds on the test bacteria which may have resulted in an underestimation of the mutagenic potencies of some of the compounds. The pattern of mutagenic activity of the hydrazines in the different tester strains indicates that more than one mechanism of action may be involved in the mutagenicity.