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Biomedical subjects

R L Crowell

Publications and source records attributed to R L Crowell.

5 recordsLinked to original sources
PubMed1979

Survival of coxsackievirus B3 under diverse environmental conditions.

The survival of coxsackievirus B3 was studied under various conditions of incubation. The comparative study demonstrated that coxsackievirus B3 was stable for 24h (less than 0.4-log decrease in titer) when suspended at neutral pH (6 or 23 degrees C) in the presence of 0.25% bovine serum albumin in saline regardless of whether the preparations were subjected to evaporation. Bovine serum albumin provided increased stability to the virus for each of the conditions tested. At 37 degrees C, evaporation greatly reduced the virus infectivity between 6 and 20 h of incubation. Nevertheless, coxsackievirus B3 was found to be stable for at least 24 h under conditions similar to those of a household environment, and its presence represents a potential biohazard to nonimmune persons. These data provide a rationale for using coxsackievirus B3 as a model for investigating the role of environmental surfaces in the transmission of enteroviral diseases.

Culture Media↗
PubMed1979

Stabilization of "A" particles of coxsackievirus B3 by a HeLa cell plasma membrane extract.

Previous studies in our laboratory showed that HeLa cell plasma membranes were recovered from sucrose gradients in two major bands and that the heavier band possessed a putative inhibitor of uncoating of coxsackievirus B3. It has now been found that the mechanism of inhibition is the stabilization of "A" particles against inactivation at 37 degrees C. [3H]uridine-labeled virions converted to A particles by band 4, the heavier band, were four times more stable at 37 degrees C than those produced by band 3. Partially purified A particles from both bands were equally unstable. It was found that the stabilizing factor was extractable by saline from band 4 and remained soluble after centrifugation (109,000 X g for 2 h). Addition to A particles of this soluble factor isolated from either band 4 or band 3 stabilized the A particles. The stabilizing factor could not be replaced by an extract from band 3 or by bovine serum albumin. Thus, the finding that the membrane factor inhibits virus uncoating by stabilizing A particles against spontaneous disruption at 37 degrees C focuses attention on an inherent problem associated with defining receptor-mediated virus uncoating.

Cell Membrane↗
PubMed1975

Elution and uncoating of Coxsackievirus B3 by isolated HeLa cell plasma membranes.

Plasma membranes isolated from HeLa cells on discontinuous sucrose gradients were assayed for their capacity to elute and uncoat coxsackievirus B3 at 37 C. Because the viral receptors are limited to the surface of HeLa cells, the addition of radioactively labeled virus to the cells prior to cell homogenization provided a useful marker for locating the plasma membranes during the fractionation procedure. Four bands were formed on the discontinuous sucrose gradients with approximately 70% or more of the membrane-associated viral label being recovered in the most dense bands, designated as bands 3 and 4. Bands 3 and 4 also possessed the plasma membrane marker enzymes, Na+, K+ adenosine triphosphatase and 5'-nucleotidase and revealed typical structures characteristic of plasma membranes as revealed by electron microscopy. Pelleted and washed membranes from band 3 both eluted and uncoated B3 32P-labeled virus, whereas membranes from band 4 eluted virus but failed to uncoat it. The membranes from band 4 were shown to inhibit the viral uncoating activity when mixed with membranes of band 3. Characteristically, unfractionated homogenates of cell membranes eluted but did not uncoat virus. The finding of a naturally occurring inhibitor of virus uncoating provides for the first time a way to distinguish between the membrane activities of virus elution and virus uncoating. The inhibitor remains to be characterized.

Adenosine Triphosphatases↗