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At least 937 records · Page 52Linked to original sources
PubMed1973

Proteins specified by herpes simplex virus. IX. Contiguity of host and viral proteins in the plasma membrane of infected cells.

Artificial mixtures of plasma membrane vesicles produced by microcavitation from infected and uninfected cells band at the same density on isopycnic centrifugation in sucrose density gradient. However, after reaction with antiviral antibody, the density of the infected cell plasma membrane vesicles increases, and the infected and uninfected cell membranes are quantitatively separable on isopycnic centrifugation. Plasma membrane vesicles prepared from cells doubly labeled before and after infection with radioactive amino acids and reacted with antibody banded at a high density. Polyacrylamide gel electropherograms show that the vesicles reacted with antibody consist of both host- and virus-specific membrane proteins. Microcavitation does not disrupt viral envelopes since infectivity is not affected by this procedure. We conclude that viral and cellular proteins in the plasma membrane preparations are contiguous.

Amino Acids↗
PubMed1970

Defective virions of reovirus.

When purified preparations of stock reovirus, type 3, were digested with chymotrypsin, the virions were converted into two different types of particle. These new particles could be separated from each other by isopycnic centrifugation in cesium chloride gradients. One particle banded at a buoyant density of 1.43 g/cm(3), the other at a density of 1.415 g/cm(3). The former particle is termed the heavy (H) particle, the latter is the light (L) particle. The ratio of H/L particles varied between 0.5 and 0.25 in various purified preparations of virus. In electron micrographs, both H and L particles had the appearance and dimensions of viral cores. H particles were infectious for L cells. When plaques formed by stock virus, or by H particles, were picked and propagated in L cells, the majority of the clones gave rise only to H particles on chymotrypsin digestion. On continued serial passage of the clones, virions containing L particles again appeared in the progeny. The simplest explanation of these results was that stock virus was comprised of two populations of virions. One type of virion which contained H particles was infectious, whereas the other, which contained L particles, was not itself infectious and could replicate only in cells coinfected with an H particle virion. Added weight was given to this hypothesis by two observations. First, a small but definite separation of H and L virions could be achieved by isopycnic centrifugation in a gradient of cesium chloride. Second, L particles and virions containing L particles were both shown to lack the largest of the ten segments of double-stranded ribonucleic acid genome. Thus, L particle virions have defective genomes.

Acrylates↗
PubMed1978

Demonstration of specific storage material within cutaneous nerves in metachromatic leukodystrophy.

Biopsies of clinically unaltered skin from a 5-year-old girl with metachromatic leukodystrophy were studied by light, fluorescence and analytical electron microscopy. Investigation of hematoxylin-eosin-stained sections revealed no pathologic changes, but a brown metachromatic material was found within cutaneous nerves after acetic acid cresyl violet staining of frozen sections. In semithin Epon sections small dermal nerve fascicles contained endoneural deposits, which proved to correspond with the typical prismatic, dense or Tuffstein bodies described in other organs of patients suffering from MLD. The inclusions were located mainly within Schwann cells and exhibited a bright orange fluorescence after trypaflavine-phosphotungstic acid-treatment as well as a high electron density in serial ultrathin sections. Energy dispersive X-ray microanalysis revealed a high sulfur content in the membrane bound granules. These findings demonstrate the presence of the specific storage material also within cutaneous nerves in MLD and thus suggest skin biopsies as an addiitonal and simple diagnostic acid in this disease.

Child, Preschool↗
PubMed1972

Chemical and physical characteristics of the deoxycholate-soluble and magnesium-reaggregated membrane nicotinamide adenine dinucleotide (reduced form) oxidase of Bacillus megaterium.

The inactive components of the nicotinamide adenine dinucleotide (reduced form) (NADH) oxidase present in the 0.4% deoxycholate-soluble fraction obtained from Bacillus megaterium KM membranes were reaggregated into active NADH oxidase by dilution in the presence of Mg(2+). The reaggregated oxidase was different from the original membrane with respect to sedimentation behavior in a sucrose gradient and morphological appearance. The deoxycholate-insoluble portion of the membrane had membrane-like structure whereas the reaggregated oxidase appeared to be a filamentous aggregate of small particles. The reaggregated oxidase and the deoxycholate-insoluble membrane residue were similar to the original membrane with respect to total protein and total lipid content. The inactive components of the NADH oxidase system exist in deoxycholate as two molecular species which were separable by sucrose density gradient centrifugation or gel filtration in deoxycholate-containing solutions. Both components and dilution in the presence of Mg(2+) were necessary for restoration of oxidase activity. The smaller-molecular-weight component contained all of the NADH-2,6-dichlorophenolindophenol oxidoreductase activity of the original membrane.

Bacillus megaterium↗
PubMed1971

Biological, physical, and chemical properties of Eastern equine encephalitis virus. I. Purification and physical properties.

A new purification procedure was adopted for Eastern equine encephalitis virus which does not subject the virus to pelleting at any stage. Three- to 4-liter volumes were passed through a diethylaminoethyl cellulose column. The virus-containing fractions were banded on a sucrose cushion and finally concentrated in an isopycnic band in a linear sucrose gradient. This method reduced the volume 1,000-fold with a concomitant increase in viral titer, i.e., better than 90% recovery. Numerous criteria have been used to establish that this viral preparation was essentially free from cellular debris and nonviral material. Physical studies on this purified viral product were initiated. The sedimentation coefficient as determined by band sedimentation was 240S, the buoyant density in sucrose was 1.18 g/cc, and the diameter of the virus was 54 nm. From the diameter and the buoyant density it was possible to calculate the molecular weight of a spherical particle. In this case, the calculated molecular weight for Eastern equine encephalitis virus was 58 x 10(6) daltons.

Amino Acids↗
PubMed1969

Microtubules in brain homogenates.

Microtubules from neurons are preserved in homogenates of mammalian brain by medium containing organic solvents at acidic pH. By means of negative staining and electron microscopy, the relative concentration of microtubules in suspensions can be assayed. Microtubules from brain have a filamentous and substructure.

Animals↗
PubMed1986

Light and electron microscopic study on complex carbohydrates in the testis of Salamandra salamandra L. (Amphibia, Urodela).

The distribution of complex carbohydrates was studied in the testis of the European fire salamander, Salamandra salamandra, by light- and electron-microscopic methods. The basal laminae and fibrous structures in the connective tissue between the lobules are PAS-positive. After alcianblue staining (at pH = 2.8), acid mucopolysaccharides could be demonstrated in steroid hormone-producing cells in the interstitial tissue between lobules containing spermatids, spermatozoa, and lobules after spermiation, as well as in most of the Sertoli cells in lobules after spermiation. In all spermatogenic stages from secondary spermatocytes to mature sperms, dictyosome-like structures and flat vesicles showed a distinct contrast enhancement, as did parts of the acrosome after treatment with the phosphotungstic acid-chromic acid method for electron microscopy.

Acrosome↗
PubMed1980

[Nomarski interference microscopy: use of combined contrast uranyl acetate-PTA for in vitro observation of monolayer cultures].

In this paper is reported a technique for the fixation and staining of cellular monolayers in vitro for observation by the interferential microscope according to Nomarski. This method includes the use of glutaraldehyde at 2.5% in phosphate buffer 0.1 M pH 7.5 and the sequential use of Uranil acetate (0.9% in absolute alcohol) and phosphtungstic acid (1% in absolute alcohol). This study has been conducted on 3 cellular lines: PC12 (rat pheocromocytoma), R.P.C. (rat pineal cells), primary culture obtained from a human carcinoma of the uterus.

Acetates↗
PubMed1992

Antithrombotic and thrombolytic activity of sulodexide in rats.

We evaluated the ability of sulodexide, an extracted glycosaminoglycan, to prevent thrombus formation and to reduce a stabilized thrombus in a rat venous thrombosis model (vena cava ligature). Injection of sulodexide 10 min before induction of venous stasis, prevented thrombus formation in a dose-dependent manner (median effective dose 0.55 mg/kg). When given to rats with 6-h-old thrombi, sulodexide caused a marked reduction in thrombus size which reached 70% after 2 h with the highest dose tested (2 mg/kg). The effect of sulodexide on established thrombi appears to be due, at least in part, to a fibrinolysis-mediated mechanism, since it was significantly inhibited by epsilon-aminocaproic acid, a well-known antifibrinolytic drug. Treatment with sulodexide did not noticeably affect plasma levels of plasminogen activator and its specific inhibitor. We also showed that fluorescein-labelled sulodexide, when given to animals with 6-h-old thrombi, was present within the thrombi harvested 2 h later, but was then absent from blood. The fluorescence was mainly located in areas filled with amorphous material, that was identified as fibrin by staining with phosphotungstic acid-hematoxylin. No fluorescein-labelled material could be detected in rats treated with fluorescein alone. These findings indicate that, besides preventing venous thrombus formation, sulodexide is able to promote thrombus dissolution by a mechanism that is partly related to local fibrinolysis stimulation.

Animals↗
PubMed1975

Colorimetric, enzymatic, and liquid-chromatographic methods for serum uric acid compared.

We describe high-performance liquid chromatography in conjunction with electrochemical detection as a possible reference method for serum uric acid. Separation was effected on a column packed with "Vydac" strong anion-exchange resin, with use of a detection potential of +0.80 V vs. an Ag/AgCl reference electrode. Results were linearly related to concentration up to 1.0 g/liter, and no interferences were seen. Assay of human sera gave within-run and day-to-day coefficients of variation of 0.83% and 1.1%, respectively; analytical recoveries averaged 100%. Comparison of the new procedure (x) with the phosphotungstate and uricase methods (y) showed the following linear regression and correlation coefficients for results: y equal 0.963x + 0.219 (r = 0.995), and y = 0.991x + 0.165 (r = 0.999), respectively. As compared to these methods, the procedure we describe is more accurate, because of the selective detection system based on retention time and redox potential. Samples can be analyzed at the rate of 20/h. No deproteinization is required.

Chromatography, Ion Exchange↗
PubMed1971

Characterization of two psychrophilic Pseudomonas bacteriophages isolated from ground beef.

Characterization studies were performed on two psychrophilic phages which were isolated from ground beef samples. Phage inactivation by exposure to heat, low pH, osmotic shock conditions, and freezing showed that these two isolates were different. One-step growth experiments indicated that one isolate had a burst size five times as large (500) and a latent period two times as long (4 hr) as the other when tested at 7 C. Nucleic acid type was 2-deoxyribonucleic acid for both. Electron micrographs showed one to belong to Bradley's phage group A and the other to phage group C.

Animals↗
PubMed1971

Structure and development of viruses as observed in the electron microscope. XI. Entry and uncoating of herpes simplex virus.

Two morphologically distinct types of capsids are described. The dense capsid appeared to be disrupted near the cellular membrane with release of core material. The light capsid was more stable and was frequently encountered close to the nucleus, where empty capsids were also found. Pretreatment of cells before infection with either puromycin or actinomycin D markedly decreased the percentage of empty capsids. It is suggested that the two types of capsids play different roles in the process of initiating infection. One (the dense capsid) releases deoxyribonucleic acid (DNA) shortly after entry. This DNA is transcribed into a virus-specific ribonucleic acid, which codes for an enzyme capable of altering the permeability of the second type of capsid (the light capsid). In proximity to the nucleus, the infectious DNA then escapes without gross disruption of the capsid.

Adsorption↗
PubMed1970

Electrn microscopic observations on virus-like particles associated with SH antigen.

The structural aspects of SH antigen-containing particles were investigated. These studies confirmed the existence of a large spherical particle (ca. 43 nm) and smaller (ca. 20 nm) rod- and sphere-shaped particles. The large particle consists of an outer and inner membrane and a core of "nucleic acid" as seen by positive staining techniques. The outer membrane of the large particle appears to be similar to that of the 20-nm diameter spheres and rods known to possess the SH antigen.

Acute Disease↗
PubMed1970

Early events in the infection of permissive cells with simian virus 40: adsorption, penetration, and uncoating.

The early events in the interaction of simian virus 40 (SV40) with permissive cells were investigated. Evidence is presented that 30 min after infection intact virions penetrate the nuclei of infected cells. The uncoating of the virus is carried out in the nuclei with a complete dissociation of the viral genome from the protein coat. Opening of the circular parental deoxyribonucleic acid (DNA), i.e., conversion of component I to component II of SV40 DNA, takes place after uncoating, followed by the appearance of a new component sedimenting faster than component I at alkaline pH.

Adsorption↗