Application of horse-radish peroxidase linked chemiluminescence to determine the production mechanism of Shiga-like toxins by E. coli O157:H7

2007 ◽  
Author(s):  
Shu-I Tu ◽  
Joseph Uknalis ◽  
Andrew Gehring ◽  
Yiping He
Keyword(s):  
Production Mechanism ◽  
Horse Radish Peroxidase ◽  
E Coli

scholarly journals Macromolecule Permeability in Rodent Intestine following Thermal Injury and Lipopolysaccharide Challenge

2013 ◽  
Vol 2013 ◽  
pp. 1-7
Author(s):  
Peirong Yu ◽  
Edward A. Carter
Keyword(s):  
Barrier Function ◽  
Thermal Injury ◽  
Blood Perfusion ◽  
Severe Trauma ◽  
Proximal Jejunum ◽  
Horse Radish Peroxidase ◽  
E Coli ◽  
Intestinal Hypoperfusion ◽  
Major Route

The barrier function of the intestinal mucosa may be lost during stress such as severe trauma and sepsis. The present study utilized a multicannulated (jugular vein, proximal jejunum, thoracic duct, and portal vein) rat model of burn (30% body surface area (TBSA)) and endotoxemia (E. coli lipopolysaccharide (LPS) infused via the jugular cannula) to investigate in vivo barrier function to macromolecules with different sizes and the route for their transport (horse radish peroxidase (HRP) and 14C-polyethylene glycol (PEG)-4,000 infused via jejunal cannula). In burn rats, mucosa uptakes of HRP and PEG increased 3 h after their intraluminal infusion compared to the controls. Studies with intravenous 111In-IgG infusion showed that its recovery in small intestine was decreased after burn and LPS infusion, indicating that blood perfusion to intestine was compromised. The present study suggests that (1) burn and endotoxemia increase intestinal permeability to macromolecules; (2) the portal blood may be the major route of transport for molecules up to sizes of 4,000 during burn but not endotoxemia; and (3) intestinal hypoperfusion could be one of the factors that contribute to increased gut permeability in severe burn trauma and sepsis.

Endotoxin Alteration of the Mucopolysaccharide Blood Vessel Coating in Rats

1974 ◽  
Vol 32 ◽  
pp. 148-149
Author(s):  
D. E. Philpott ◽  
A. Takahashi
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Salivary Gland ◽  
Carotid Artery ◽  
Basement Membrane ◽  
Coronary Vessels ◽  
Ruthenium Red ◽  
E Coli ◽  
Old Rats ◽  
The Brain

Two month, eight month and two year old rats were treated with 10 or 20 mg/kg of E. Coli endotoxin I. P. The eight month old rats proved most resistant to the endotoxin. During fixation the aorta, carotid artery, basil arartery of the brain, coronary vessels of the heart, inner surfaces of the heart chambers, heart and skeletal muscle, lung, liver, kidney, spleen, brain, retina, trachae, intestine, salivary gland, adrenal gland and gingiva were treated with ruthenium red or alcian blue to preserve the mucopolysaccharide (MPS) coating. Five, 8 and 24 hrs of endotoxin treatment produced increasingly marked capillary damage, disappearance of the MPS coating, edema, destruction of endothelial cells and damage to the basement membrane in the liver, kidney and lung.

Ribosome Structural Organization

1974 ◽  
Vol 32 ◽  
pp. 332-333
Author(s):  
James A. Lake
Keyword(s):  
Ribosomal Proteins ◽  
Structural Organization ◽  
Three Dimensional ◽  
Small Subunit ◽  
Structural Studies ◽  
X Ray Diffraction ◽  
Specific Antibodies ◽  
X Ray ◽  
E Coli ◽  
Complete Sequences

The understanding of ribosome structure has advanced considerably in the last several years. Biochemists have characterized the constituent proteins and rRNA's of ribosomes. Complete sequences have been determined for some ribosomal proteins and specific antibodies have been prepared against all E. coli small subunit proteins. In addition, a number of naturally occuring systems of three dimensional ribosome crystals which are suitable for structural studies have been observed in eukaryotes. Although the crystals are, in general, too small for X-ray diffraction, their size is ideal for electron microscopy.

Permeability of Endometrial Blood Vessels to Exogenous Horse Radish Peroxidase in Mice. Electron Microscope Study

1973 ◽  
Vol 31 ◽  
pp. 562-563
Author(s):  
M.C. Castillo-Jessen ◽  
A. González-Angulo
Keyword(s):  
Electron Microscopy ◽  
Blood Vessels ◽  
Electron Microscope Study ◽  
Ultrastructural Level ◽  
Low Molecular Weight ◽  
Horse Radish Peroxidase ◽  
Intravascular Injection ◽  
Normal Morphology ◽  
Functional Implications ◽  
Low Molecular Weight Proteins

Information regarding the normal morphology of uterine blood vessels at ultrastructural level in mammals is scarce Electron microscopy studies dealing with endometrial vasculature despite the functional implications due to hormone priming are not available. Light microscopy observations with combined injection of dyes and microradiography along with histochemical studies does not enable us to know the detailed fine structure of the possible various types of blood vessels in this tissue. The present work has been designed to characterize the blood vessels of endometrium of mice as well as the behavior of the endothelium to injection of low molecular weight proteins during the normal estrous cycle in this animal. One hundred and forty female albino mice were sacrificed after intravascular injection of horse radish peroxidase (HRP) at 30 seconds, 5, 15, 30 and 60 minutes.

Sites of Adsorption of the Bacteriophages T3 and T5 on the Wall of Escherichia Coli B.

1967 ◽  
Vol 25 ◽  
pp. 96-97
Author(s):  
Manfred E. Bayer
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Electron Microscope ◽  
Uranyl Acetate ◽  
E Coli ◽  
Receptor Sites ◽  
Rigid Cell Wall ◽  
Host Cell Surface ◽  
Bacterial Viruses ◽  
Escherichia Coli B

Bacterial viruses adsorb specifically to receptors on the host cell surface. Although the chemical composition of some of the cell wall receptors for bacteriophages of the T-series has been described and the number of receptor sites has been estimated to be 150 to 300 per E. coli cell, the localization of the sites on the bacterial wall has been unknown.When logarithmically growing cells of E. coli are transferred into a medium containing 20% sucrose, the cells plasmolize: the protoplast shrinks and becomes separated from the somewhat rigid cell wall. When these cells are fixed in 8% Formaldehyde, post-fixed in OsO4/uranyl acetate, embedded in Vestopal W, then cut in an ultramicrotome and observed with the electron microscope, the separation of protoplast and wall becomes clearly visible, (Fig. 1, 2). At a number of locations however, the protoplasmic membrane adheres to the wall even under the considerable pull of the shrinking protoplast. Thus numerous connecting bridges are maintained between protoplast and cell wall. Estimations of the total number of such wall/membrane associations yield a number of about 300 per cell.

Emigration of neutrophils in the central nervous system

1983 ◽  
Vol 41 ◽  
pp. 788-789
Author(s):  
John L.Beggs ◽  
John D. Waggener ◽  
Wanda Miller ◽  
Jane Watkins
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Osmium Tetroxide ◽  
White Blood Cells ◽  
Arterial Perfusion ◽  
E Coli ◽  
Intracisternal Injection ◽  
The Central Nervous System ◽  
Brain And Spinal Cord ◽  
Interendothelial Junctions

Studies using mesenteric and ear chamber preparations have shown that interendothelial junctions provide the route for neutrophil emigration during inflammation. The term emigration refers to the passage of white blood cells across the endothelium from the vascular lumen. Although the precise pathway of transendo- thelial emigration in the central nervous system (CNS) has not been resolved, the presence of different physiological and morphological (tight junctions) properties of CNS endothelium may dictate alternate emigration pathways.To study neutrophil emigration in the CNS, we induced meningitis in guinea pigs by intracisternal injection of E. coli bacteria.In this model, leptomeningeal inflammation is well developed by 3 hr. After 3 1/2 hr, animals were sacrificed by arterial perfusion with 3% phosphate buffered glutaraldehyde. Tissues from brain and spinal cord were post-fixed in 1% osmium tetroxide, dehydrated in alcohols and propylene oxide, and embedded in Epon. Thin serial sections were cut with diamond knives and examined in a Philips 300 electron microscope.

Use of Uranium-Labeled Antibodies for Electron Microscopic Localization of Various Antigens in Mammalian Cells

1967 ◽  
Vol 25 ◽  
pp. 136-137
Author(s):  
J. P. Petrali ◽  
E. J. Donati ◽  
L. A. Sternberger
Keyword(s):  
Endoplasmic Reticulum ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Specific Antiserum ◽  
Electron Microscopic ◽  
E Coli ◽  
Cytoplasmic Membranes ◽  
Viral Progeny ◽  
Ultrathin Sections ◽  
Electron Microscopic Localization

Specific contrast is conferred to subcellular antigen by applying purified antibodies, exhaustively labeled with uranium under immunospecific protection, to ultrathin sections. Use of Seligman’s principle of bridging osmium to metal via thiocarbohydrazide (TCH) intensifies specific contrast. Ultrathin sections of osmium-fixed materials were stained on the grid by application of 1) thiosemicarbazide (TSC), 2) unlabeled specific antiserum, 3) uranium-labeled anti-antibody and 4) TCH followed by reosmication. Antigens to be localized consisted of vaccinia antigen in infected HeLa cells, lysozyme in monocytes of patients with monocytic or monomyelocytic leukemia, and fibrinogen in the platelets of these leukemic patients. Control sections were stained with non-specific antiserum (E. coli).In the vaccinia-HeLa system, antigen was localized from 1 to 3 hours following infection, and was confined to degrading virus, the inner walls of numerous organelles, and other structures in cytoplasmic foci. Surrounding architecture and cellular mitochondria were unstained. 8 to 14 hours after infection, antigen was localized on the outer walls of the viral progeny, on cytoplasmic membranes, and free in the cytoplasm. Staining of endoplasmic reticulum was intense and focal early, and weak and diffuse late in infection.

Structural similarity of ribosomes from E. coli and A. salina

1978 ◽  
Vol 36 (2) ◽  
pp. 242-243
Author(s):  
M. Boublik ◽  
R.M. Wydro ◽  
W. Hellmann ◽  
F. Jenkins
Keyword(s):  
Ribosomal Proteins ◽  
Direct Evidence ◽  
Structural Similarity ◽  
Carbon Support ◽  
Artemia Salina ◽  
Uranyl Acetate ◽  
Biological Assays ◽  
E Coli ◽  
And Function ◽  
Fine Carbon

Ribosomes are ribonucleoprotein particles necessary for processing the genetic information of mRNA into proteins. Analogy in composition and function of ribosomes from diverse species, established by biochemical and biological assays, implies their structural similarity. Direct evidence obtained by electron microscopy seems to be of increasing relevance in understanding the structure of ribosomes and the mechanism of their role in protein synthesis.The extent of the structural homology between prokaryotic and eukaryotic ribosomes has been studied on ribosomes of Escherichia coli (E.c.) and Artemia salina (A.s.). Despite the established differences in size and in the amount and proportion of ribosomal proteins and RNAs both types of ribosomes show an overall similarity. The monosomes (stained with 0.5% aqueous uranyl acetate and deposited on a fine carbon support) appear in the electron micrographs as round particles with a diameter of approximately 225Å for the 70S E.c. (Fig. 1) and 260Å for the 80S A.s. monosome (Fig. 2).

RNA polymerase: Preparation and structural analysis of helical polymers

1984 ◽  
Vol 42 ◽  
pp. 152-153
Author(s):  
E. Loren Buhle ◽  
Pamela Rew ◽  
Ueli Aebi
Keyword(s):  
Structural Analysis ◽  
Rna Polymerase ◽  
Molecular Level ◽  
X Ray Diffraction ◽  
Helical Polymers ◽  
X Ray ◽  
E Coli ◽  
The Past ◽  
Ordered Arrays ◽  
Low Ionic Strength

While DNA-dependent RNA polymerase represents one of the key enzymes involved in transcription and ultimately in gene expression in procaryotic and eucaryotic cells, little progress has been made towards elucidation of its 3-D structure at the molecular level over the past few years. This is mainly because to date no 3-D crystals suitable for X-ray diffraction analysis have been obtained with this rather large (MW ~500 kd) multi-subunit (α2ββ'ζ). As an alternative, we have been trying to form ordered arrays of RNA polymerase from E. coli suitable for structural analysis in the electron microscope combined with image processing. Here we report about helical polymers induced from holoenzyme (α2ββ'ζ) at low ionic strength with 5-7 mM MnCl2 (see Fig. 1a). The presence of the ζ-subunit (MW 86 kd) is required to form these polymers, since the core enzyme (α2ββ') does fail to assemble into such structures under these conditions.

An Ultrastructural Study of Chronic Pyelonephritis in Macaca Arctoides

1976 ◽  
Vol 34 ◽  
pp. 310-311
Author(s):  
T.W. Smith ◽  
J.A. Roberts ◽  
B.J. Martin
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Ultrastructural Study ◽  
Effective Means ◽  
Chronic Pyelonephritis ◽  
Macaca Arctoides ◽  
Left Ureter ◽  
E Coli ◽  
Transmission Electron ◽  
Sizeable Number

Chronic pyelonephritis is one of the most common diseases of the kidney and accounts for a sizeable number of cases of renal insufficiency in man, however its pathogenesis requires further elucidation. Transmission electron microscopy may serve as a uniquely effective means of observing details of the nature of this disease. The present paper describes preliminary results of an ultrastructural study of chronic pyelonephritis in Macaca arctoides (stumptail monkey).The infection was induced in these experiments in a retrograde fashion by means of a unilateral catheterization of the left ureter whereby an innoculum of 10 cc of broth containing approximately 2 billion E. coli per cc and radio-opaque dye were injected under pressure (mimicing vesico-ureteric reflux).

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