scholarly journals PHAGE FORMATION IN STAPHYLOCOCCUS MUSCAE CULTURES

1950 ◽  
Vol 34 (2) ◽  
pp. 251-277 ◽  
Author(s):  
Winston H. Price
Keyword(s):  
Aspartic Acid ◽  
Phage Particle ◽  
Burst Size ◽  
Synthetic Medium ◽  
Acid Synthesis ◽  
Active Particle ◽  
Multiple Infection ◽  
Experimental Conditions ◽  
Virus Particles ◽  
Infected Cells

1. A strain of S. muscae which requires a substance present in certain acid-hydrolyzed proteins (AHPF) for virus liberation when singly infected in Fildes' synthetic medium no longer needs this substance when multiply infected. 2. In the absence of the AHPF under conditions of multiple infection the amount of phage released is approximately equal to the number of infecting particles between two to ten. Over ten particles per cell has no further effect on the yield of virus. 3. The experimental evidence indicates that it is the phage particle and not some other component in the lysate which can replace the AHPF. 4. The minimum latent period and rise period of cells singly infected in the presence of the AHPF and multiply infected in the absence of the AHPF are the same. 5. The desoxynucleic acid synthesis of cells, infected with a very few virus particles in the presence of excess AHPF and multiply infected with ten particles in the absence of the AHPF, occurs at approximately the same rate, with both infected samples synthesizing about the same amount of desoxynucleic acid and liberating the same yields of virus. 6. A strain of S. muscae which requires aspartic acid for virus synthesis when singly infected does not need this substance when multiply infected, the burst size under the latter conditions depending upon the multiplicity of infection between 3 to 12 particles per cell. 7. The data indicate that the virus released from multiply infected cells in the absence of added AHPF or aspartic acid is newly synthesized virus and not the original infecting particles. 8. The phage particle contains the AHPF and aspartic acid. 9. As a tentative working hypothesis, it is assumed that the AHPF and aspartic acid for phage formation under conditions of multiple infection, in the absence of added AHPF, or of aspartic acid, are contributed by the original infecting particles. 10. Ultraviolet-inactivated phage is adsorbed to the host cell and kills the cell although little virus is released under the experimental conditions. 11. Ultraviolet-inactivated phage particles, if added before the active particle is adsorbed, will greatly inhibit the liberation of new virus particles; but does not do so if added a few minutes after the active particle has been adsorbed. 12. Under the experimental conditions, reactivation of phage when present in multiply infected cells does not occur; and such ultraviolet-inactivated phage cannot serve as a source of the AHPF or aspartic acid, although the AHPF can be liberated from such inactivated particles by acid hydrolysis. 13. The results are discussed in relation to Luria's experiments with ultraviolet-treated phage and to his "gene pool" hypothesis of phage formation.

scholarly journals PHAGE FORMATION IN STAPHYLOCOCCUS MUSCAE CULTURES

1950 ◽  
Vol 34 (2) ◽  
pp. 231-250 ◽  
Author(s):  
Winston H. Price
Keyword(s):  
Aspartic Acid ◽  
Burst Size ◽  
Synthetic Medium ◽  
Growth Curves ◽  
Acid Synthesis ◽  
Virus Protein ◽  
Complete Medium ◽  
Multiplication Rate ◽  
One Step ◽  
Step Growth

1. Four strains of Staphylococcus muscae have been isolated which differ in their growth rates and phage syntheses in Fildes' synthetic medium. 2. Two of the strains when singly infected cannot release phage in Fildes' synthetic medium unless a substance present in certain acid-hydrolyzed proteins is added to the medium. One of these strains also requires other substance(s) present in acid-hydrolyzed proteins in order to grow in Fildes' medium. 3. The two strains which do not require the addition of the phage-stimulating factor have been found either to synthesize this substance, or one similar to it. One of these strains will not grow in Fildes' medium unless substance(s) present in acid-hydrolyzed proteins is added to the medium. 4. The purified acid-hydrolyzed protein factor necessary for virus liberation does not affect the multiplication rate of uninfected S. muscae cells in Fildes' synthetic medium. 5. The substance is not needed for the adsorption or the invasion of the host cell by the virus. In the absence of the factor, the virus is adsorbed to the cell and "kills" it. 6. An analysis carried out by means of the one-step growth curve technique has indicated that the substance is not concerned simply with the mechanism of virus release, but is necessary for some initial stage in virus synthesis. 7. With one bacterial strain not requiring the AHPF, aspartic acid had to be present at least during the minimum latent period for the cell to form virus. 8. In the absence of aspartic acid, the virus was adsorbed to the cell and killed it, but no virus was released from singly infected bacteria. 9. If the cells were grown in a medium containing aspartic acid and then resuspended in the medium minus aspartic acid, no virus was released, although such cells contained at least two times the amount of aspartic acid necessary for the burst size in the complete medium. 10. Aspartic acid, a constituent of the virus particle, appears from an analysis of one-step growth curves to take part in the initial phase of phage synthesis. 11. The effect of amino acids on virus formation is discussed in relation to the time sequence of virus protein and desoxyribonucleic acid synthesis.

scholarly journals Nuclear Synthesis of Cytoplasmic Ribonucleic Acid in Amoeba proteus

1959 ◽  
Vol 6 (2) ◽  
pp. 203-206 ◽  
Author(s):  
David M. Prescott
Keyword(s):  
Ribonucleic Acid ◽  
Rna Synthesis ◽  
Synthetic Medium ◽  
Acid Synthesis ◽  
Amoeba Proteus ◽  
Food Vacuole ◽  
Yeast Cells ◽  
Starvation Period ◽  
Experimental Conditions ◽  
Food Vacuoles

The enucleation technique has been applied to Amoeba proteus by several laboratories in attempts to determine whether the cytoplasm is capable of nucleus-independent ribonucleic acid synthesis. This cell is very convenient for micrurgy, but its use requires a thorough starvation period to eliminate the possibility of metabolic influence by food vacuoles and frequent washings and medium renewal to maintain asepsis. In the experiments described here, amoebae were starved for periods of 24 to 96 hours, cut into nucleated and enucleated halves, and exposed to either C-14 uracil, C-14 adenine, C-14 orotic acid, or a mixture of all three. When the starvation period was short (less than 72 hours), organisms (especially yeast cells) contained within amoeba food vacuoles frequently showed RNA synthesis in both nucleated and enucleated amoebae. When the preperiod of starvation was longer than 72 hours, food vacuole influence was apparently negligible, and a more meaningful comparison between enucleated and nucleated amoebae was possible. Nucleated cells incorporated all three precursors into RNA; enucleated cells were incapable of such incorporation. The experiments indicate a complete dependence on the nucleus for RNA synthesis. The conflict with the experimental results of others on this problem could possibly stem from differences in culture conditions, starvation treatment, or experimental conditions. For an unequivocal answer in experiments of this design, ideally the cells should be capable of growth on an entirely synthetic medium under aseptic conditions. The use of a synthetic medium (experiments with A. proteus are done under starvation conditions) would permit, moreover, a more realistic comparison of metabolic capacities of nucleated and enucleated cells.

Further Observations on the Virus of Epizootic Diarrhea of Infant Mice. An Electron Microscopic Study

1967 ◽  
Vol 25 ◽  
pp. 110-111
Author(s):  
W. G. Banfield ◽  
G. Kasnic ◽  
J. H. Blackwell
Keyword(s):  
Electron Microscope ◽  
Infected Cell ◽  
Microscopic Study ◽  
Intestinal Epithelium ◽  
Ultrastructural Study ◽  
Osmium Tetroxide ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Virus Particles ◽  
Infected Cells

An ultrastructural study of the intestinal epithelium of mice infected with the agent of epizootic diarrhea of infant mice (EDIM virus) was first performed by Adams and Kraft. We have extended their observations and have found developmental forms of the virus and associated structures not reported by them.Three-day-old NLM strain mice were infected with EDIM virus and killed 48 to 168 hours later. Specimens of bowel were fixed in glutaraldehyde, post fixed in osmium tetroxide and embedded in epon. Sections were stained with uranyl magnesium acetate followed by lead citrate and examined in an updated RCA EMU-3F electron microscope.The cells containing virus particles (infected) are at the tips of the villi and occur throughout the intestine from duodenum through colon. All developmental forms of the virus are present from 48 to 168 hours after infection. Figure 1 is of cells without virus particles and figure 2 is of an infected cell. The nucleus and cytoplasm of the infected cells appear clearer than the cells without virus particles.

Studies of a Defective Measles Virus

1968 ◽  
Vol 26 ◽  
pp. 208-209
Author(s):  
R. M. McCombs ◽  
M. Benyesh-Melnick ◽  
J. P. Brunschwig
Keyword(s):  
Inclusion Bodies ◽  
Measles Virus ◽  
Giant Cells ◽  
Helical Structures ◽  
Thin Sections ◽  
Virus Particles ◽  
Extracellular Virus ◽  
Culture Cells ◽  
Infected Cells ◽  
Eosinophilic Inclusions

Measles virus is an agent that is capable of replicating in a number of different culture cells and generally causes the formation of multinucleated giant cells. As a result of infection, virus is released from the cells into the culture fluids and reinfection can be initiated by this cell-free virus. The extracellular virus has been examined by negative staining with phosphotungstic acid and has been shown to be a rather pleomorphic particle with a diameter of about 140 mμ. However, no such virus particles have been detected in thin sections of the infected cells. Rather, the only virus-induced structures present in the giant cells are eosinophilic inclusions (intracytoplasmic or intranuclear). These inclusion bodies have been shown to contain helical structures, resembling the nucleocapsid observed in negatively stained preparations.

scholarly journals Zeolite NaP1 Functionalization for the Sorption of Metal Complexes with Biodegradable N-(1,2-dicarboxyethyl)-D,L-aspartic Acid

Materials ◽  
2021 ◽  
Vol 14 (10) ◽  
pp. 2518
Author(s):  
Dorota Kołodyńska ◽  
Yongming Ju ◽  
Małgorzata Franus ◽  
Wojciech Franus
Keyword(s):  
Experimental Data ◽  
Aspartic Acid ◽  
Ph Value ◽  
Complexing Agents ◽  
X Ray Diffraction ◽  
Experimental Conditions ◽  
Modified Zeolite ◽  
X Ray ◽  
Slow Release Fertilizers ◽  
Pseudo Second Order

The possibility of application of chitosan-modified zeolite as sorbent for Cu(II), Zn(II), Mn(II), and Fe(III) ions and their mixtures in the presence of N-(1,2-dicarboxyethyl)-D,L-aspartic acid, IDHA) under different experimental conditions were investigated. Chitosan-modified zeolite belongs to the group of biodegradable complexing agents used in fertilizer production. NaP1CS as a carrier forms a barrier to the spontaneous release of the fertilizer into soil. The obtained materials were characterized by Fourier transform infrared spectroscopy (FTIR); surface area determination (ASAP); scanning electron microscopy (SEM-EDS); X-ray fluorescence (XRF); X-ray diffraction (XRD); and carbon, hydrogen, and nitrogen (CHN), as well as thermogravimetric (TGA) methods. The concentrations of Cu(II), Zn(II), Mn(II), and Fe(III) complexes with IDHA varied from 5–20 mg/dm3 for Cu(II), 10–40 mg/dm3 for Fe(III), 20–80 mg/dm3 for Mn(II), and 10–40 mg/dm3 for Zn(II), respectively; pH value (3–6), time (1–120 min), and temperature (293–333 K) on the sorption efficiency were tested. The Langmuir, Freundlich, Dubinin–Radushkevich, and Temkin adsorption models were applied to describe experimental data. The pH 5 proved to be appropriate for adsorption. The pseudo-second order and Langmuir models were consistent with the experimental data. The thermodynamic parameters indicate that adsorption is spontaneous and endothermic. The highest desorption percentage was achieved using the HCl solution, therefore, proving that method can be used to design slow-release fertilizers.

scholarly journals Regulation of Host Ribonucleic Acid Synthesis in Bacteriophage T3-infected Cells

1974 ◽  
Vol 249 (6) ◽  
pp. 1787-1791
Author(s):  
S.P. Mahadik ◽  
B. Dharmgrongartama ◽  
P.R. Srinivasan
Keyword(s):  
Ribonucleic Acid ◽  
Acid Synthesis ◽  
Infected Cells

scholarly journals Coat protein and RNAs of Cole latent virus are not present within chloroplasts of Chenopodium quinoa-infected cells

2003 ◽  
Vol 28 (1) ◽  
pp. 84-88 ◽  
Author(s):  
Priscila Belintani ◽  
José O. Gaspar
Keyword(s):  
Electron Microscopy ◽  
Coat Protein ◽  
Chenopodium Quinoa ◽  
Latent Virus ◽  
Percoll Gradient ◽  
Northern Blots ◽  
Virus Particles ◽  
Ultrastructural Studies ◽  
Particle Aggregates ◽  
Infected Cells

Cole latent virus (CoLV), genus Carlavirus, was studied by electron microscopy and biochemical approaches with respect both to the ultrastructure of the Chenopodium quinoa infected cells and to its association with chloroplasts. The CoLV was observed to be present as scattered particles interspersed with membranous vesicles and ribosomes or as dense masses of virus particles. These virus particles reacted by immunolabelling with a polyclonal antibody to CoLV. Morphologically, chloroplasts, mitochondria and nuclei appeared to be unaltered by virus infection and virus particles were not detected in these organelles. However, virus particle aggregates were frequently associated with the outer membrane of chloroplasts and occasionally with peroxisomes. Chloroplasts were purified by Percoll gradient, and the coat protein and virus-associated RNAs were extracted and analyzed by Western and Northern blots respectively. Coat protein and CoLV-associated RNAs were not detected within this organelle. The results presented in this work indicate that the association CoLV/chloroplasts, observed in the ultrastructural studies, might be a casual event in the host cell, and that the virus does not replicate inside the organelle.

scholarly journals Human Cytomegalovirus TR Strain Glycoprotein O Acts as a Chaperone Promoting gH/gL Incorporation into Virions but Is Not Present in Virions

2009 ◽  
Vol 84 (5) ◽  
pp. 2597-2609 ◽  
Author(s):  
Brent J. Ryckman ◽  
Marie C. Chase ◽  
David C. Johnson
Keyword(s):  
Endothelial Cells ◽  
Human Cytomegalovirus ◽  
Null Mutant ◽  
Virus Particles ◽  
Extracellular Virus ◽  
Biochemical Studies ◽  
Infected Cells ◽  
Low Passage ◽  
Er Export ◽  
Strain Ad169

ABSTRACT Human cytomegalovirus (HCMV) produces the following two gH/gL complexes: gH/gL/gO and gH/gL/UL128-131. Entry into epithelial and endothelial cells requires gH/gL/UL128-131, and we have provided evidence that gH/gL/UL128-131 binds saturable epithelial cell receptors to mediate entry. HCMV does not require gH/gL/UL128-131 to enter fibroblasts, and laboratory adaptation to fibroblasts results in mutations in the UL128-131 genes, abolishing infection of epithelial and endothelial cells. HCMV gO-null mutants produce very small plaques on fibroblasts yet can spread on endothelial cells. Thus, one prevailing model suggests that gH/gL/gO mediates infection of fibroblasts, while gH/gL/UL128-131 mediates entry into epithelial/endothelial cells. Most biochemical studies of gO have involved the HCMV lab strain AD169, which does not assemble gH/gL/UL128-131 complexes. We examined gO produced by the low-passage clinical HCMV strain TR. Surprisingly, TR gO was not detected in purified extracellular virus particles. In TR-infected cells, gO remained sensitive to endoglycosidase H, suggesting that the protein was not exported from the endoplasmic reticulum (ER). However, TR gO interacted with gH/gL in the ER and promoted export of gH/gL from the ER to the Golgi apparatus. Pulse-chase experiments showed that a fraction of gO remained bound to gH/gL for relatively long periods, but gO eventually dissociated or was degraded and was not found in extracellular virions or secreted from cells. The accompanying report by P. T. Wille et al. (J. Virol., 84:2585-2596, 2010) showed that a TR gO-null mutant failed to incorporate gH/gL into virions and that the mutant was unable to enter fibroblasts and epithelial and endothelial cells. We concluded that gO acts as a molecular chaperone, increasing gH/gL ER export and incorporation into virions. It appears that gO competes with UL128-131 for binding onto gH/gL but is released from gH/gL, so that gH/gL (lacking UL128-131) is incorporated into virions. Thus, our revised model suggests that both gH/gL and gH/gL/UL128-131 are required for entry into epithelial and endothelial cells.

scholarly journals A CYTOCHEMICAL DESCRIPTION OF THE MULTIPLICATION OF MENGOVIRUS IN L-929 CELLS

1962 ◽  
Vol 12 (1) ◽  
pp. 1-15 ◽  
Author(s):  
Richard M. Franklin
Keyword(s):  
Phase Contrast ◽  
Basic Protein ◽  
Virus Production ◽  
Cytological Change ◽  
Virus Particles ◽  
Contrast Microscopy ◽  
Infected Cells ◽  
Cytological Changes ◽  
Perinuclear Area

A correlation of cytochemical changes with virus production has been studied in L cells infected with Mengovirus. After a latent period of about 2 hours, virus was produced rapidly, reaching maximum titers of up to 12,000 particles per cell in 6 to 8 hours. The earliest cytological change was in the nucleus and consisted of a slight condensation of chromatin. There is no evidence, however, for the multiplication of either the viral RNA or protein in the nucleus. RNA, of high molecular weight, accumulated in the perinuclear area of the cytoplasm and was later found in inclusions. The perinuclear RNA was digestible with RNase and may be located in or on ribosomes. The inclusion RNA was resistant to RNase but could be removed by pepsin or potassium permanganate; it is probably in completed virus particles. Viral antigen was first observed in a perinuclear location and later in the above-mentioned inclusions. Although the viral protein contains appreciable amounts of arginine and lysine, it is not a basic protein of the histone type. Phase-contrast microscopy of living cells clearly demonstrated the role of the inclusions in release of virus from infected cells. A comparison is made between these cytological changes in Mengo-infected cells and those which have been found by other workers in polio-infected cells. There are many very similar changes.

scholarly journals STRUCTURE OF TYPE 5 ADENOVIRUS

1963 ◽  
Vol 118 (2) ◽  
pp. 295-306 ◽  
Author(s):  
Wesley C. Wilcox ◽  
Harold S. Ginsberg
Keyword(s):  
Virus Particle ◽  
Density Gradient ◽  
Infectious Virus ◽  
Relative Proportion ◽  
Specific Antigen ◽  
Equilibrium Density ◽  
Virus Particles ◽  
Infected Cells ◽  
Specific Antigens ◽  
Virus Antigens

Type 5 adenovirus was purified by fluorocarbon (freon 113) treatment followed by banding in a CsCl equilibrium density gradient. This method permitted separation of virus from normal host cell materials and virus-specific soluble antigens. Virus banded in CsCl with a mean bouyant density of 1.3349 gm/cm3. The three virus-specific soluble antigens (group- and type-specific antigens and toxin) banded together with a mean bouyant density of 1.2832 gm/cm3. The group-specific antigen was the predominant antigen of the purified virus particle, whereas the group- and type-specific antigens were present in equal titers in the antigen band. Infectious virus particles were inactivated by prolonged dialysis at pH 10.5. Centrifugation of inactivated virus preparations in a CsCl equilibrium density gradient resulted in separation of virus DNA from specific antigen: the antigens banded with a mean bouyant density of 1.2832 gm/cm3 and the DNA sedimented to the bottom of the tube. The predominant antigen derived from purified virus particles was the group-specific antigen and it was in the same relative proportion to the type-specific antigen as measured in intact particles. The antigens derived from disrupted virus were immunologically identical with the soluble virus antigens present in infected cells.

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