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H G Aaslestad

Publications and source records attributed to H G Aaslestad.

12 recordsLinked to original sources
PubMed1997

Sabia virus incident at Yale University.

An incident involving a human exposure to a newly isolated arenavirus, Sabia virus, in the Yale Arbovirus Research Unit occurred at Yale University on August 8, 1994. A senior-level visiting research scientist was exposed to Sabia virus while purifying the virus from a large volume of tissue culture fluid. The exposure resulted in development of a Sabia virus infection followed by recovery of the patient. The incident resulted in a comprehensive review by a Yale faculty committee and an external expert committee. As a result, a number of new practices and procedures were added to Yale's biosafety policy for investigating infectious agents in BL-3 facilities.

Air Microbiology↗
PubMed1977

Lipids of rabies virus and BHK-21 cell membranes.

The lipid composition of highly purified Flury strain of rabies virus (HEP) propagated in BHK-21 cells in a chemically defined medium was observed to be 6.7% neutral lipids, 15.8% phospholipids, and 1.5% glycolipids. In the virion, phosphatidylethanolamine, phosphatidylcholine, and sphingomyelin were the most abundant phospholipids, accounting for 90% of the total, and the molar ratio of cholesterol to phospholipid was 0.48. Uninfected BHK-21 cell membranes were obtained by nitrogen cavitation techniques and separated by density gradient centrifugation, and the membranes were assayed for purity using 5'-nucleotidase, cytochrome oxidase, and reduced nicotinamide adenine dinucleotide phosphate diaphorase activities. Lipids of the plasma membrane were enriched in cholesterol, phosphatidylcholine, and phosphatidylethanolamine. In contrast, membranes of the endoplasmic reticulum were enriched in phosphatidylcholine, but contained smaller amounts of phosphatidylethanolamine and sphingomyelin. Comparison of the fatty acyl chains of virus and membranes from uninfected cells revealed the virion to have the lowest ratio of C18:1 to C18:0 (1.771), compared with values of about 3.0 for the plasma membrane and endoplasmic reticulum. Total polyenoic fatty acids were enriched in the plasma membrane, whereas the virus contained higher amounts of total saturates than either of the two membrane preparations. Analysis of the polar and neutral lipid fractions as well as the acyl chain analysis suggests the virion has a lipid composition that is intermiediate to that of the plasma membrane and endoplasmic reticulum and is consistent with the view that numerous viral particles are synthesized de novo by not utilizing a preexisting membrane template. From the ratio of cholesterol to phospholipid of 0.48, we calculated that 1.92 X 10(5) molecules of lipid would cover 4.14 X 10(4) nm2 in the form of a bilayer. Considerations of the molecular dimensions of the rabies envelope (total surface area, 5 X 10(4) nm2) as a bilayer suggest that some penetration of lipids by envelope proteins (M and G) is necessary.

Animals↗
PubMed1975

Partial purification and biochemical characterization of herpesvirus saimiri.

HVS propagated in Vero cells was labelled with 3H-leucine and purified by isopycnic flotation in sucrose. Electron microscopic examination revealed both complete and incomplete HVS. No significant contamination with cellular debris was noted. Polyacrylamide gel electrophoresis analysis of solubilized virus revealed at least 12 proteins varying in molecular weight from 8 000 to 200 000 daltons.

Centrifugation, Isopycnic↗
PubMed1972

Immunogenicity of rabies virus inactivated by -propiolactone, acetylethyleneimine, and ionizing irradiation.

Ionizing radiation, beta-propiolactone, and acetylethyleneimine were compared for their ability as virus-inactivating agents for the preparation of rabies vaccine. Each agent reduced viral infectivity exponentially; ionizing radiation also destroyed viral hemagglutinin. The vaccine prepared by ionizing radiation was equal or superior to that prepared by beta-propiolactone in its ability to protect mice from rabies infection. The acetylethyleneimine-treated vaccine was a less potent immunogen.

Acetates↗
PubMed1972

Recovery of protective activity in rabies virus vaccines concentrated and purified by four different methods.

Rabies vaccines concentrated by ultrafiltration, zinc acetate precipitation, ammonium sulfate precipitation, or aluminum phosphate gel adsorption were compared with respect to recovery of protective activity and purity, as measured by protective activity per mg of protein. Vaccine obtained by ammonium sulfate precipitation had a better recovery rate and a higher purity than those prepared by the other methods. Potent vaccines were also obtained by the zinc acetate precipitation and aluminum phosphate gel adsorption methods, whereas ultrafiltration was the least satisfactory method from the standpoint of vaccine purity. Chromatography of virus concentrated by ultrafiltration on a cellulose ion exchange column reduced the level of nonviral proteins. The protective activity data obtained for the vaccines examined in these experiments were found to correlate with the vaccine's complement fixation titer per mg of protein.

Acetates↗
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↗
PubMed1971

Comparison of the ribonucleic acid polymerases of two rhabdoviruses, Kern Canyon virus and vesicular stomatitis virus.

A ribonucleic acid (RNA)-dependent RNA polymerase has been demonstrated in Kern Canyon virus (KCV) particles. The RNA product of the KCV polymerase hybridizes to KCV viral RNA. The properties of this viral enzyme have been characterized and compared with those of vesicular stomatitis virus (VSV). RNA polymerases from both viruses require similar conditions of temperature, pH, and detergent and magnesium concentrations for maximal synthesis of RNA. The RNA polymerase contained in the virion of KCV was more dependent on the presence of a sulfhydryl agent than was the VSV enzyme. Under optimal conditions, the specific activity of the VSV polymerase is about twenty-five times as great as that of KCV.

Animals↗
PubMed1968

Fractionation of Eastern equine encephalitis virus by density gradient centrifugation in CsCl.

When partially purified Eastern equine encephalitis (EEE) virus was centrifuged to equilibrium in CsCl, three virus specific bands were observed. A hemagglutinin was detected at a buoyant density of 1.18 g/cm(3). Infectious EEE virus banded in two positions; most of the virus banded at 1.20 g/cm(3) and a lesser amount banded at 1.22 to 1.23 g/cm(3). Analysis of radioactive profiles of CsCl-fractionated EEE virus labeled with either (32)PO(4) or (3)H-uridine suggested that the hemagglutinin was stripped from the intact EEE virion. The viral origin of the hemagglutinin was verified by inhibition with specific antiserum. Attempts to differentiate between infectious EEE virus of the different buoyant densities showed that the denser particle was neither a virus contaminant nor a density mutant. No evidence was obtained to indicate that the denser particle was an immature form of EEE virus. The two infectious EEE species obtained after CsCl fractionation were indistinguishable antigenically. Furthermore, unfractionated as well as CsCl-fractionated EEE virus sedimented at about 260S in sucrose gradients. These results together with the results of rebanding experiments suggested that the denser EEE species (1.23 g/cm(3)) results from a salt (CsCl)-induced alteration or breakdown of the EEE virion (1.20 g/cm(3)), and that it arises as the hemagglutinin is stripped from the surface of the EEE virion.

Arboviruses↗