Biomedical subjects
C MORGAN
Publications and source records attributed to C MORGAN.
Intranuclear crystals of herpes simplex virus observed with the electron microscope.
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Use of the electron microscope in the study of intracellular virus.
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Intracellular crystals associated with viral development.
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An evaluation of host cell changes accompanying viral multiplication as observed in the electron microscope.
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Electron microscopic and histochemical studies of an unusual crystalline protein occurring in cells infected by type 5 adenovirus.
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A correlated histochemical and electron microscopic study of the intranuclear crystalline aggregates of adenovirus (RI-APC virus) in HeLa cells.
HeLa cells in tissue cultures infected with types 3, 4, or 7 of adenovirus (RI-APC virus) were studied in order to correlate certain histochemical and electron microscopic findings. Adjacent thin (ca. 0.05 micro) and thick (2-4 micro) sections of osmium-fixed, methacrylate-embedded cells were cut; by mapping the sections the same cells could be identified with both the electron and the light microscope. Intranuclear crystalline aggregates seen with the electron microscope to be composed of ordered arrays of viral particles were found by means of the Feulgen reaction to contain DNA. DNA is therefore assumed to be a constituent of the viral particle. The virus appeared to develop from an osmiophilic Feulgen-negative matrix. Displacement of nuclear chromatin occurred during this process. A Feulgen-azure staining method was found to permit clear distinction between viral and nuclear (host) DNA in thick sections.
Structure and development of viruses observed in the electron microscope. III. Influenza virus.
Rods and spheres believed to represent viral particles were observed at the free surface of entodermal cells of the chorioallantoic membrane 6 to 44 hours after infection. Although occasional short rods revealed poorly defined internal bodies, the majority, as well as all the longer rods (filaments), exhibited no visible internal structure. The spheres presumed to lie central to the plane of section contained an inner body 20 to 22 mmicro in diameter. Both forms possessed a dense, sharply defined limiting membrane 30 A thick and a diffuse external coat of lesser density. Where superimposition within the section was minimal, the viral particles were separated by a relatively constant distance. Measured to include this spacing, on the assumption that it reflected the presence of a component of the outer coat, the diameters of a majority of the rods were 50 to 60 mmicro, whereas the spheres averaged 60 to 70 mmicro. The rods appeared to form by a process of extrusion from the cell wall and became detached either singly or in bundles of variable length. The spheres seemed to differentiate at the cell surface and to acquire the inner body, limiting membrane, and outer coat as they migrated through the membrane of the host cell. No characteristic changes were seen in the nuclei or adjacent cytoplasm, and recognizable viral particles were never encountered in these areas of the cell. No support was obtained for the assumption that the spheres developed primarily by segmentation of the rods. It is suggested that the spherical form of the virus is the elemental infectious unit and that the filamentous form is largely or completely non-infective.
Some effects of the microtome knife and electron beam on methacrylate-embedded thin sections.
A technique for the examination of specimens at low electron beam intensity has been presented. Sections micrographed with this technique showed numerous knife scratches and frequently contained bands running parallel to the knife edge. Banding with an average spacing of 0.2 micro appeared to result from periodic distortion produced by impact of the knife. At the beam intensities customarily employed, differential sublimation and probably flow of the methacrylate resulted in obliteration of the bands and all but the deepest knife scratches. In addition, changes in the size, shape, and orientation of certain structures were noted. Artifacts resulting from incineration or sublimation of tissue components fixed in formalin were illustrated, and the suggestion was made that such instability to the electron beam accounted in part for the differences observed in osmium- and formalin-fixed tissues. The deformation revealed in serial sections was discussed, and it was pointed out that shortening in the axis perpendicular to the knife edge was associated with elongation in the axis parallel to the cutting edge, the elongation usually occurring locally without change in the width of the section. It was noted that the material causing contamination of the surface of sections during examination exhibited no structure but caused progressive loss of contrast.
Use of serial sections to delineate the structure of Porthetria dispar virus in the electron microscope.
Consecutive serial sections of polyhedra obtained from gipsy moth larvae infected with P. dispar virus revealed bundles of viral rods scattered and oriented at random within the polyhedral body. Each bundle was entirely surrounded by a dense, sharply defined membrane. The rods measured 18 to 22 mmicro in diameter and averaged 280 mmicro in length. No spherical viral particles were encountered. The effects of variable compression and periodic distortion of the sections on the appearance of the virus are discussed.
A possible means of identifying certain viruses by their structure.
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The macromolecular paracrystalline lattice of insect viral polyhedral bodies demonstrated in ultrathin sections examined in the electron microscope.
Thin sections of polyhedra obtained from gipsy moth larvae infected with P. dispar virus and from silkworm larvae infected with B. mori virus revealed viral particles contained within a pseudohexagonal, macromolecular, paracrystalline lattice. The gipsy moth virus occurs in bundles of one to eight rods enclosed by a limiting membrane. The particles of the silkworm virus, although generally occurring singly, also possess a limiting membrane. The macromolecules appear to be dense, discrete particles when cross-sectioned and to form dense bands by superimposition when longitudinally or obliquely sectioned at certain angles. Calculations of macromolecular size have been made.
Serial sections of vaccinia virus examined at one stage of development in the electron microscope.
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The appearance of herpes simplex, influenza, vaccinia and fowlpox viruses within sectioned chorioallantoic membranes examined in the electron microscope.
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Preliminary clinical observations on oxamycin: a new antibiotic.
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Structure and development of viruses observed in the electron microscope. II. Vaccinia and fowl pox viruses.
Vaccinia and fowl pox viruses were visualized by the electron microscope in sections of infected chorioallantoic membrane of chicken embryos. The viruses were of similar structure and size, averaging 200 x 300 mmicro with considerable individual variation. Intracytoplasmic viral particles contained a dense, nucleus-like body (nucleoid) separated from granular material (viroplasm) by a zone of lesser density. They were enclosed by a single membrane. Near the surface of the host cell and in the extracellular space the particles consisted of a central body of variable shape and density enclosed by a double membrane. The initial sites of development were confined to the cytoplasm of the host cell. Before release from the host cell the viral nucleoids appeared to enlarge and to occupy a central position within the particle, which became enclosed by a double limiting membrane. The brick-shaped forms found after removal of the embedding plastic from thick sections indicated that drying caused characteristic distortion of certain viral particles.
Structure and development of viruses as observed in the electron microscope. I. Herpes simplex virus.
Herpes simplex virus was visualized by the electron microscope in sections of infected chorioallantoic membrane of chicken embryos. Removal of the embedding methacrylate from relatively thick sections permitted large numbers of viral particles to be seen but caused extensive alteration of cellular components as well as variable distortion of viral structure. This distortion was characterized by disruption of particles and loss of central bodies, resulting in ring or empty shell forms. An inner structure of the virus was revealed in ultrathin sections from which the embedding plastic was not removed. The nuclei of infected cells contained small, dense, primary bodies (30 to 40 mmicro in diameter) as well as slightly larger and less dense particles (40 to 50 mmicro in diameter) surrounded by a single membrane (70 to 100 mmicro in diameter). In the cytoplasm most of the particles possessed a double outer membrane (120 to 130 mmicro in diameter). It is suggested that the initial site of viral development is restricted to the nucleus where primary bodies form and become enclosed by a single outer membrane. Upon release into the cytoplasm these particles appear to acquire a second outer membrane and presumably represent the mature virus.
Internal structure in virus particles.
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