Mechanisms of poliovirus inactivation by the direct and indirect effects of ionizing radiation.
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Biomedical subjects
Publications and source records attributed to R L Ward.
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The effect of detergents on the stability of enteric viruses was found to be highly dependent on pH. This was demonstrated primarily with two ionic detergents, sodium dodecyl sulfate (an anionic detergent) and dodecyltrimethylammonium chloride (a cationic detergent). Both detergents were shown to be potent virucidal agents for reovirus, but the effects of sodium dodecyl sulfate were minimal near neutrality and much more pronounced at low than at high pH values. Dodecyltrimethylammonium chloride was extremely virucidal at high pH's but had little observable effect on reovirus stability at low pH values. In contrast, both detergents protected enteroviruses against heat at neutral and alkaline pH's. However, as was found with reovirus, sodium dodecyl sulfate was extremely virucidal at pH values below 5, even when the virus samples were incubated in ice. At different pH's the effects of detergents on the stabilities of coliphages T4, f1, and Q beta were qualitatively similar to those found with reovirus. Differences in viral stability in these experiments appeared to be due to the effects of pH on the ionic states of the viral capsid proteins.
Association with sludge solids did not allow poliovirus to enter cells other than through the normal receptor-mediated route. This implies that the infectivities of sludge-encapsulated virions are masked until their exposure permits binding to cell surface receptors.
Methods for inactivating bacterial viruses in serum were developed through the use of heat and ionizing radiation, and the effects of these treatments on the growth rates of cultured cells were tested. Viruses chosen for this study were the radiation-resistant bacteriphage f2 and heat-resistant phage T4. The viabilities of these phages were reduced more than 2 and 4 orders of magnitude, respectively, by a treatment at 60 degrees C for 30 min followed by 420 krads of ionizing radiation. Simultaneous application of heat and radiation caused a considerably greater reduction in viability of both phages in serum, but also caused a significant decrease in the growth rates of L cells in medium supplemented with serum treated in this manner. Treatment of serum with these same doses but given in the sequential fashion of heat followed by radiation caused little or no change in the growth rates of L cells. Finally, it was found that simultaneous treatment of serum with these doses of heat and radiation had little effect on the growth rates of either HeLa or Chinese hamster cells.
A case of immunoblastic lymphadenopathy which underwent transformation into immunoblastic sarcoma is reported. A 64-year-old man presented with a rash, generalised lymphadenopathy, and hepatosplenomegaly. A cervical lymph node removed at biopsy showed the features of immunoblastic lymphadenopathy with the presence of heavy chain classes IgG, IgM, and IgA and both kappa and lambda light chain types in the cytoplasm of the immunoblasts. No such immunoglobulins could be demonstrated in the lymph nodes obtained at necropsy when the patient died of widespread immunoblastic sarcoma. The biological evolution and histogenesis of the disease are discussed and the current literature is reviewed.
Permeability rates were determined across excised rabbit corneas for 11 steroids. Permeability coefficients for each steroid were calculated, and their logarithms were plotted against their respective log octanol-water partition coefficients. A parabolic relationship resulted, with an optimum log permeability and coefficient observed at a log Po of 2.9. From these experimental results, an improvement in ophthalmic bioavailability of dexamethasone acetate as compared to dexamethasone is predicted and correlates with literature results.
Gel formulations containing 2% pilocarpine hydrochloride were prepared from ethylene maleic anhydride, carbomer, hydroxyethylcellulose, polyacrylamide, ethylhydroxyethylcellulose, hydroxypropylcellulose, and poly(methylvinyl ether--maleic anhydride). The viscosity characteristics of each formulation were evaluated from rheograms developed at 37 degrees using a cone and plate viscometer. Single-point viscosities were determined at room temperature using a single-point rotational viscometer. Plastic viscosity parameters correlated to miosis durations in the rabbit following ophthalmic dosing of 50 microliters. Carbomer formulations varying in concentration between 0.9 and 5.0% (w/w) showed a discontinuous relationship when either yield value or plastic viscosity was plotted against miosis durations. At carbomer concentrations above 3%, miosis durations increased 1.5-fold. Above and below this range, plastic parameters did not correlate to miosis duration. It was reasoned that the increased duration was a consequence of the gel's increased yield value such that appreciable in vivo thinning of the gel does not take place with eyelid and/or eyeball movements. As a result, the residence time of the drug in the eye would be expected to increase, thus promoting an increased duration.
Anabaena cylindrica sparged with argon gas produced H2 continuously for 30 days under limited light conditions (6.0 W/m2) and for 18 days under elevated light conditions (32 W/m2) in the absence of exogenous nitrogen. The efficiency of converting visible light energy (32 W/m2) into chemical energy that is trapped as H2 ranged between 0.35 and 0.85% (approximately 13 microliter of H2 per mg [drywt] per h). Ammonium additions (0.2 mM NH4+) at various times destabilized the system and eventually suppressed H2 production completely, as compared with the control. Cultures grown with 5.0 mg of Fe3+ per liter produced H2 at a rate about twice that of cultures with 0.5 mg of Fe3+ per liter. Cultures grown at pH 7.4 produced H2 at the same initial rates as cultures that were grown at pH 9.4; however, the latter cultures continued to produce H2 after CO2 deprivation.
The abilities of ammonia and related compounds to inactivate poliovirus were compared. Compounds virucidal at pH 9.5 had the following order of activities: ethylamine greater than propylamine, dimethylamine, methylamine greater than ammonia greater than 2-methoxyethylamine.
The agent in wastewater sludge previously shown to reduce the heat required to inactivate reovirus (R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol. 34:681--688, 1977) was "separated" from other sludge components and analyzed by infrared spectroscopy. The infrared spectrum of this material was quite similar to the spectra of commercial anionic detergents, and subsequent analyses of the fractionated sludge samples revealed that anionic detergents in sludge were copurified with the virucidal activity. Further measurements on the virucidal activities of specific detergents revealed that ionic detergents reduce the heat required to inactivate reovirus, that cationic detergents are more active than anionic, and that nonionic detergents are inactive. Several detergents were also shown to protect poliovirus and other enteroviruses against inactivation by heat. These results indicate that ionic detergents are the major component in wastewater sludge that reduce the thermal stability of reovirus and, in addition, that detergents are able to protect enteroviruses against heat.
The effect of moisture content on the rates of heat inactivation of enteric viruses in wastewater sludge was determined. The protective effect of raw sludge on poliovirus previously observed (R. L. Ward, C. S. Ashley, and R. H. Moseley, Appl. Environ. Microbiol. 32:339--346, 1976) was found to be greatly enhanced in sludge dewatered by evaporation. Other enteroviruses responded in a similar fashion. This effect did not appear to be due merely to the state of dryness of the sludge samples because in humus-deficient soil, a relatively inert material, the rate of poliovirus inactivation by heat was not significantly altered through dewatering. Instead, this effect appeared to have been caused by protective substances in the sludge, such as detergents, which are concentrated through dewatering. As reported previously (R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol. 34:681-688, 1977; R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol 36:889--897, 1978) raw sludge is not protective of reovirus, but, instead, the ionic detergents in sludge cause the rate of heat inactivation of this virus to be accelerated. Dewatering of sludge, however, was found to partially reverse this virucidal effect. Evidence is presented indicating that this reversal is caused by an unidentified protective substance in sludge also concentrated through dewatering. Finally, it was shown that the effects of raw sludge on heat inactivation of poliovirus and reovirus are greatly reduced by composting, a result that correlated with the degradation of detergents.
Poliovirus inactivation by ammonia causes a slight reduction in the sedimentation coefficients of viral particles, but has no detectable effect on either the electrophoretic pattern of viral capsid proteins or the isoelectric points of inactivated particles. These virions still attach to cells, but are unable to repress host translation or stimulate the synthesis of detectable amounts of viral RNA. Although ammonia has no detectable effect on naked poliovirus RNA, it causes cleavage of this RNA when still within viral particles. Therefore, the RNA genome appears to be the only component of poliovirus significantly affected by ammonia.
A study of 43 pediatric oral surgery patients was conducted in a clinical environment. The anesthetic regimen of a minimum dose of ketamine hydrochloride, nitrous oxide-oxygen, and a local anesthetic was tested for recovery time and degree of acceptance by parents and patients. With this regimen, the average recovery time from completion of the procedure was 30 minutes. The combination of these drugs provided a rapid, smooth induction, additional time while the ketamine-induced anesthesia was dissipating, no major complications, and good acceptance by parents and patients.
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Anaerobically digested sludge contains an agent that causes irreversible inactivation of poliovirus. It has now been shown that the agent responsible for this activity is ammonia. The effect of ammonia on poliovirus appears to be typical for picornaviruses, but reovirus, an enteric virus of another group, is quite resistant to this compound. Because ammonia is not virucidal in its charged state, it expresses significant activity only at pH values greater than 8. Therefore, increasing the pH of sludge should cause rapid inactivation of indigenous picornaviruses.
Sludge is highly protective of poliovirus against both heat and ionizing radiation. This protection could be largely overcome when these two treatments were given simultaneously.
The effect of dewatering on the inactivation rates of enteric viruses in sludge was determined. For this study, water was evaporated from seeded raw sludge at 21 degrees C, and the loss of viral plaque-forming units was measured. Initial results with poliovirus showed that recoverable infectivity gradually decreased with the loss of water until the solids content reached about 65%. When the solids content was increased from 65 to 83%, a further, more dramatic decrease in virus titer of greater than three orders of magnitude was observed. This loss of infectivity was due to irreversible inactivation of poliovirus because viral particles were found to have released their RNA molecules which were extensively degraded. Viral inactivation in these experiments may have been at least partially caused by the evaporation process itself because similar effects on poliovirus particles were observed in distilled water after only partial loss of water by evaporation. Coxsackievirus and reovirus were also found to be inactivated in sludge under comparable conditions, which suggests that dewatering by evaporation may be a feasible method of inactivating all enteric viruses in sludge.