Loss of the bacterial flagellar motor switch complex upon cell lysis

Microbial Biotechnolog

Discovery of a novel inner membrane-associated bacterial structure related to the flagellar type III secretion system

Microbial Biotechnolog

Emerging techniques: Cryo-electron microscopy of vitrified biological specimens

Rapid freezing has long promised the most faithful structural presentation of biological material for electron microscopy. With the recent emergence of methods for vitrifying specimens, this promise seems about to be fulfilled

The outer-membrane, not a coat of host proteins, limits antigenicity of virulent Treponema-pallidum

Virulent Treponema pallidum reacts poorly with the specific antibodies present in human and rabbit syphilitic sera, a phenomenon often attributed to an outer coat of host serum proteins. Here we present additional evidence that the limited antigenicity of virulent organisms actually is due to a paucity of proteins in the outer membrane. Initially, we used electron microscopy to demonstrate that the outer membrane is highly susceptible to damage from physical manipulation (i.e., centrifugation and resuspension) and nonionic detergents. Organisms with disrupted outer membranes were markedly more antigenic than intact treponemes as determined by immunoelectron microscopy (IEM) with rabbit syphilitic and antiendoflagellar antisera. Data obtained with a new radioimmunoassay, designated the T. pallidum surface-specific radioimmunoassay, corroborated these IEM findings by demonstrating that the major T. pallidum immunogens are not surface exposed; the assay also was unable to detect serum proteins, including fibronectin, on the surfaces of intact organisms. Furthermore, IEM of T. pallidum on ultrathin cryosections with monospecific anti-47-kDa-immunogen antiserum confirmed the intracellular location of the 47-kDa immunogen. On the basis of these and previous findings, we proposed a new model for T. pallidum ultrastructure in which the outer membrane contains a small number of transmembrane proteins and the major membrane immunogens are anchored by lipids to the periplasmic leaflet of the cytoplasmic membrane. This unique ultrastructure explains the remarkable ability of virulent organisms to evade the humoral immune response of the T. pallidum-infected host

Electron microscopic visualization of a discrete class of giant translation units in salivary gland cells of Chironomus tentans

Lysates were made of whole salivary glands from Chironomus tentans and electron microscopic spreads prepared according to Miller and Beatty (1969). The sedimented material consisted mainly of ribosomes, most of which were present in giant polysomes which showed gradients of material protruding laterally from the ribosomes, interpreted as nascent polypeptide chains. Each giant polysome contained one such gradient. On the basis of the presence of a small group of terminal ribosomes without nascent chains at the 3' end, 19 apparently complete polysomes were selected, representing a discrete class of polysomes containing between 66 and 92 ribosomes (79 ± 7, m ± s.d.). Arguments are presented that they constitute the translation units for giant secretory proteins coded by RNA from the large Balbiani rings, BR1 and BR2

Epithelial injury induces keratocyte apoptosis: hypothesized role for the interleukin-1 system in the modulation of corneal tissue organization and wound healing

The purpose of this study was to determine the role of programmed cell death (apoptosis) in the disappearance of keratocytes beneath an epithelial debridement wound in the cornea and to investigate a potential role of interleukin-l (IL-1) in induction of apoptosis in stromal fibroblasts in vitro and keratocytes in vivo. Keratocyte and stromal fibroblast cell morphology was examined in wounded and unwounded mouse corneas using transmission electron microscopy. Nuclear DNA fragmentation was detected with the TUNEL assay for 3'-hydroxyl DNA ends. The effect of IL-1 on keratocytes in vivo was determined by microinjection of IL-1 alpha into the central corneal stroma via a limbal entry site, The in vitro effects of interleukin-1 alpha (IL-1 alpha) and interleukin-l beta (IL-1 beta) were determined with primary cultures of human corneal stromal and dermal fibroblasts. Cell shrinkage, blebbing with formation of membrane bound bodies, condensation and fragmentation of the chromatin, and DNA fragmentation, consistent with apoptosis were detected in anterior stromal keratocytes after epithelial scrape wounds. Thus, disappearance of keratocytes from the underlying stroma following epithelial debridement is mediated by apoptosis. Microinjection of IL-1 alpha into the central stroma of the mouse cornea caused a redistribution of keratocytes in the stroma via apoptosis and, possibly, negative chemotaxis. IL-1 alpha and IL-1 beta induced apoptosis in corneal stromal and dermal fibroblasts in vitro. The epithelial/endothelial-stromal IL-1 system may mediate corneal tissue organization and responses to mechanical- and pathogen-induced injury through induction of keratocyte apoptosis. Keratocyte apoptosis is likely an initiating event in wound healing following corneal surgery. We hypothesize that derangement's in this system may have a role in the pathogenesis of keratoconus and other diseases of the cornea. (C) 1996 Academic Press Limite