LYSIS OF CELL WALLS AND INTACT CELLS OF PSEUDOMONAS AERUGINOSA BY ETHYLENEDIAMINE TETRAACETIC ACID AND BY LYSOZYME

1965 ◽  
Vol 11 (2) ◽  
pp. 193-201 ◽  
Author(s):  
R. G. Eagon ◽  
K. Jane Carson
Keyword(s):  
Electron Microscopy ◽  
Pseudomonas Aeruginosa ◽  
Cell Walls ◽  
Chelating Agents ◽  
Tetraacetic Acid ◽  
Intact Cells ◽  
Divalent Metals ◽  
Ethylenediamine Tetraacetic Acid

It was shown that Pseudomonas aeruginosa strain 64 was extensively Iysed by EDTA alone and that destruction of cells was as complete as when lysozyme was used in combination with EDTA as evidenced by microscopic examinations and viability tests of the bacterial suspensions. Lysozyme alone caused no appreciable lysis. Lysozyme was not completely without effect. Cells that were preincubated with lysozyme, washed, and resuspended in the presence of EDTA lysed at the same rate and to the same extent as in a system containing both EDTA and lysozyme. Electron microscopy revealed that, after incubation with lysozyme, cells showed altered physical appearances manifested by "ballooning" and bulging of the cell walls. Incubation of cells with rhodamine-labelled lysozyme indicated that lysozyme is bound to the cellular surface and that it is readily removed by washing.There is evidence that EDTA acts through its chelation properties. Calcium was chelated in greatest quantity. EDTA was replaceable by structurally related polyacetic acid chelating agents but not by other chelating agents or by detergents. It is concluded that the binding of divalent metals, which may function to form cross-linkages, is essential for the integrity of the cell walls of P. aeruginosa.

Characterization of a protein-lipopolysaccharide complex released from cell walls of Pseudomonas aeruginosa by ethylenediaminetetraacetic acid

1969 ◽  
Vol 15 (7) ◽  
pp. 743-748 ◽  
Author(s):  
S. W. Rogers ◽  
H. E. Gilleland Jr. ◽  
R. G. Eagon
Keyword(s):  
Electron Microscopy ◽  
Pseudomonas Aeruginosa ◽  
Cell Walls ◽  
Ethylenediaminetetraacetic Acid ◽  
Gradient Centrifugation ◽  
Hollow Spheres ◽  
Gel Filtration ◽  
A Chain ◽  
Ultracentrifugal Analysis

Results from analytical ultracentrifugal analysis, Sephadex gel filtration, isopycnic density-gradient centrifugation, and polyacrylamide disc-gel electrophoresis revealed that ethylenediaminetetraacetic acid liberated a protein–lipopolysaccharide complex from cell walls of Pseudomonas aeruginosa with an estimated molecular weight of not less than 160 000 and probably about one million. Electron microscopy of this complex revealed spherules and rodlets. The diameter of the former was approximately 70 ± 10 Å while the dimensions of the latter were 70 ± 10 Å × 200 ± 50 Å. The rodlets appeared to be composed of three or more spherules arranged in a chain-like fashion. Electron microscopy of protein-free lipopolysaccharide revealed predominantly hollow spheres from 300 Å to 1500 Å in diameter, morphologically resembling membrane sacculi. It is proposed that the protein–lipopolysaccharide complex, but not the protein-free lipopolysaccharide, is representative of the in situ form of native endotoxin.

scholarly journals BINDING OF CALCIUM AND MAGNESIUM BY THE CONTRACTILE ELEMENTS

1955 ◽  
Vol 38 (6) ◽  
pp. 735-742 ◽  
Author(s):  
Emil Bozler
Keyword(s):  
Mechanical Properties ◽  
Muscle Fibers ◽  
Tetraacetic Acid ◽  
Salt Solutions ◽  
Dilute Solutions ◽  
Partial Loss ◽  
Inorganic Pyrophosphate ◽  
Divalent Metals ◽  
Ethylenediamine Tetraacetic Acid ◽  
Calcium And Magnesium

Using a technique for determining Ca and Mg based on Schwarzenberg's method of titration with ethylenediamine tetraacetic acid (EDTA), it was found that glycerol-extracted muscle fibers contain on the average 0.58 millimole Ca and 0.55 millimole Mg per kg. muscle. The fibers take up additional Ca or Mg from dilute solutions of these metals, but in KCl solutions, the excess is exchanged for K ions. Inorganic pyrophosphate (PP) removes part of the bound Mg, no Ca; EDTA removes predominantly Ca, but never more than about one-half the total amount. These results are discussed in relation to previous observations on the effects of PP and EDTA on mechanical properties and contractility of extracted muscle fibers. After the partial loss of bound divalent metals, muscle fibers swell in dilute salt solutions; they also contract slightly and become more translucent.

Ectodesmata as Revealed By Freeze-Etch Preparations of Plant Epidermal Cell Walls

1973 ◽  
Vol 31 ◽  
pp. 468-469
Author(s):  
N.C. Lyon ◽  
W. C. Mueller
Keyword(s):  
Electron Microscopy ◽  
Acetic Acid ◽  
Nitric Acid ◽  
Oxalic Acid ◽  
Light Microscopy ◽  
Epidermal Cell ◽  
Cell Walls ◽  
Plant Viruses ◽  
Plant Leaves ◽  
Thin Sections

Schumacher and Halbsguth first demonstrated ectodesmata as pores or channels in the epidermal cell walls in haustoria of Cuscuta odorata L. by light microscopy in tissues fixed in a sublimate fixative (30% ethyl alcohol, 30 ml:glacial acetic acid, 10 ml: 65% nitric acid, 1 ml: 40% formaldehyde, 5 ml: oxalic acid, 2 g: mecuric chloride to saturation 2-3 g). Other workers have published electron micrographs of structures transversing the outer epidermal cell in thin sections of plant leaves that have been interpreted as ectodesmata. Such structures are evident following treatment with Hg++ or Ag+ salts and are only rarely observed by electron microscopy. If ectodesmata exist without such treatment, and are not artefacts, they would afford natural pathways of entry for applied foliar solutions and plant viruses.

Silicification of wood-cell walls

1994 ◽  
Vol 52 ◽  
pp. 428-429
Author(s):  
S. E. Keckler ◽  
D. M. Dabbs ◽  
N. Yao ◽  
I. A. Aksay
Keyword(s):  
Electron Microscopy ◽  
Cell Wall ◽  
Cell Walls ◽  
Lignin Content ◽  
Resin Composite ◽  
Middle Lamella ◽  
Cellulose Fibers ◽  
Complex Structures ◽  
Rich Region ◽  
Inorganic Precursors

Cellular organic structures such as wood can be used as scaffolds for the synthesis of complex structures of organic/ceramic nanocomposites. The wood cell is a fiber-reinforced resin composite of cellulose fibers in a lignin matrix. A single cell wall, containing several layers of different fiber orientations and lignin content, is separated from its neighboring wall by the middle lamella, a lignin-rich region. In order to achieve total mineralization, deposition on and in the cell wall must be achieved. Geological fossilization of wood occurs as permineralization (filling the void spaces with mineral) and petrifaction (mineralizing the cell wall as the organic component decays) through infiltration of wood with inorganics after growth. Conversely, living plants can incorporate inorganics into their cells and in some cases into the cell walls during growth. In a recent study, we mimicked geological fossilization by infiltrating inorganic precursors into wood cells in order to enhance the properties of wood. In the current work, we use electron microscopy to examine the structure of silica formed in the cell walls after infiltration of tetraethoxysilane (TEOS).

Actinomycetes in discolored wood of living silver maple

1981 ◽  
Vol 59 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Robert A. Blanchette ◽  
John B. Sutherland ◽  
Don L. Crawford
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Carbon Source ◽  
Cell Walls ◽  
Hydroxybenzoic Acid ◽  
Liquid Media ◽  
Streptomyces Strain ◽  
Culture Techniques ◽  
Silver Maple ◽  
Scanning Electron

The greenish-brown margin of discolored wood in three living silver maple trees, Acer saccharinum L., was examined by scanning electron microscopy and microbiological culture techniques. Micrographs of xylem vessels revealed filamentous structures; some of them appeared to be actinomycetous hyphae. Actinomycetes identified as Streptomyces parvullus Waksman & Gregory, S. sparsogenes Owen, Dietz & Camiener, and a third Streptomyces strain were isolated repeatedly from discolored wood of each tree. These isolates grew in liquid media in the presence of 0.1% (w/v) concentrations of several phenols. Although other phenols included in the test were not substantially degraded, p-hydroxybenzoic acid was utilized as a carbon source by S. parvullus. All three actinomycetes inhibited growth of selected wood-inhabiting fungi when paired on malt agar. When inoculated on sterilized sapwood and discolored wood from silver maple, the actinomycetes colonized vessel walls and occlusions, but were not observed to decay cell walls.

scholarly journals THE INCORPORATION OF RADIOACTIVE PROLINE INTO CULTURED CELLS

1968 ◽  
Vol 39 (3) ◽  
pp. 698-715 ◽  
Author(s):  
H. W. Israel ◽  
M. M. Salpeter ◽  
F. C. Steward
Keyword(s):  
Electron Microscopy ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Cultured Cells ◽  
Initial Period ◽  
Special Problem ◽  
General Concept ◽  
Cell Wall Protein ◽  
Quiescent Cells ◽  
Growth Induction

Cultured carrot explants, stimulated to grow rapidly in a medium containing coconut milk, were labeled with radioactive proline. After an initial period of absorption (8 hr for proline-3H; 24 hr for proline-14C) the tissue was allowed to grow for a further period of 6 days in a similar medium free from the radioactivity. Samples were prepared for electron microscopy and radioautography at the end of the absorption period and also after the further growth. The distribution of the products from the radioactive proline in the cells is shown by high-resolution radioautography and is rendered quantitative for the different regions of the cells. The results show that the combined label, which was present in the form of proline and the hydroxyproline derived from it, was all in the protoplasm, not in the cell walls. Any combined label that appeared to be over the cell walls is shown to be due to scatter from adjacent cytoplasmic sites. Initially the radioactivity was concentrated in nuclei, even more so in nucleoli, but it subsequently appeared throughout the ground cytoplasm and was also concentrated in the plastids. The significance of these observations for the general concept of a plant cell wall protein and for the special problem of growth induction in otherwise quiescent cells is discussed.

scholarly journals The extraction of cell walls of Pseudomonas aeruginosa with aqueous phenol. The insoluble residue and material from the aqueous layers

1967 ◽  
Vol 105 (2) ◽  
pp. 759-765 ◽  
Author(s):  
K. Clarke ◽  
G. W. Gray ◽  
D. A. Reaveley
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Wall ◽  
Cell Walls ◽  
Lipid A ◽  
Diaminopimelic Acid ◽  
Insoluble Residue ◽  
Muramic Acid ◽  
Gram Negative ◽  
Gram Negative Organisms ◽  
Residue Fraction

1. The insoluble residue and material present in the aqueous layers resulting from treatment of cell walls of Pseudomonas aeruginosa with aqueous phenol were examined. 2. The products (fractions AqI and AqII) isolated from the aqueous layers from the first and second extractions respectively account for approx. 25% and 12% of the cell wall and consist of both lipopolysaccharide and muropeptide. 3. The lipid part of the lipopolysaccharide is qualitatively similar to the corresponding material (lipid A) from other Gram-negative organisms, as is the polysaccharide part. 4. The insoluble residue (fraction R) contains sacculi, which also occur in fraction AqII. On hydrolysis, the sacculi yield glucosamine, muramic acid, alanine, glutamic acid and 2,6-diaminopimelic acid, together with small amounts of lysine, and they are therefore similar to the murein sacculi of other Gram-negative organisms. Fraction R also contains substantial amounts of protein, which differs from that obtained from the phenol layer. 5. The possible association or aggregation of lipopolysaccharide, murein and murein sacculi is discussed.

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