Topical ocular anesthetics in ocular irritancy testing: a review.

The routine use of topical anesthetics to alleviate discomfort associated with in vivo ocular irritancy testing has been advocated. This review provides information about the adverse effects of topical ocular anesthetics and answers the questions: are topical anesthetics practical and effective in ocular irritancy protocols, is long-term use contraindicated, will topical anesthetics alter the response of a test substance, and are there significant side-effects which might cause pain and suffering in test animals? There was no evidence to support the use of a specific topical anesthetic. Further, information about using systemic analgesics or combinations with local anesthetics that would effectively alleviate discomfort associated with ocular irritancy testing without affecting test results was not found. Comprehensive studies are needed to identify the most effective combination of drugs that would ameliorate discomfort associated with ocular irritation testing.     

omega-6 and omega-3 fatty acids: monolayer packing and effects on bilayer permeability and cholesterol exchange.

It has been suggested that the polyunsaturated omega-3 fatty acid, docosahexaenoic acid (DHA), can adopt unique closely packed arrays in lipid bilayers (Glomset and Applegate. (1986) J. Lipid Res. 27, 658-680). These conformations are predicted on the basis of molecular dynamics calculations and are in contrast to the expanded conformations characteristic of omega-6 unsaturated fatty acids. It has also been suggested that close packing of omega-3 acyl chains could have a substantial affect on the physical properties of lipid bilayers (e.g. permeability). We report here some experimental tests of these predictions. Surface pressure-area experiments have been carried out on DHA and its mixtures with stearic and oleic acids. At low surface pressures DHA is more expanded than oleic acid. Extrapolation to the high surface pressures characteristic of lipid bilayers indicates that the area per molecule of DHA is only marginally less than that for oleic acid. Thus there is no compelling evidence to suggest that the average area per molecule of the omega-3 fatty acid is substantially different from the omega-6 fatty acid at high surface pressures. Experiments also show that the permeability of bilayers to glucose and the rates of dissociation of pyrenyl cholesterol from bilayers were similar for bilayers containing DHA compared to bilayers containing oleic acid or linoleic acid.     

Experimental infection of American winter flounder (Pseudopleuronectes americanus) with Glugea stephani (Microsporida)

Young-of-the-year Pseudopleuronectes americanus were captured in Sandy Hook Bay, New Jersey and maintained in aquaria at 16–19° C for 115 days in 1982 and 127 days in 1983. Half of them were experimentally exposed to Glugea stephani and the others were sham exposed. During the course of these experiments fish from both groups died as a result of G. stephani infection, demonstrating Glwgea -induced mortality in this species. Infections were found in both control and experimentally exposed fish. Mortality of Glwgea -infected fish in the exposed group was 63% while that in the controls was 28.5%.     

Analysis of trifluorothymidine-resistant (TFTr) mutants of L5178Y/TK+/- mouse lymphoma cells

Three classes of TFTr variants of L5178Y/TK+/- -3.7.2C mouse lymphoma cells can be identified--large colony (lambda), small colony (sigma), and tiny colony (tau). The sigma and lambda mutants are detectable in the routine mutagenesis assay using soft agar cloning. The tau mutants are extremely slow growing and are quantitated only in suspension cloning in microwells. Variants of all three classes have been analyzed in the process of evaluating the usefulness of the thymidine kinase locus in L5178Y/TK+/- mouse lymphoma cells for detecting induced mutational damage. 150 of 152 variants from mutagen treated cultures and 163 of 168 spontaneous mutants were TFTr when rechallenged approximately 1 week after isolation (3 weeks after induction). All of the 41 mutants assayed for enzyme activity were TK-deficient. The sigma and tau phenotypes were found to correlate with slow cellular growth rates (doubling time greater than 12 h), rather than from effects of the TFT selection or mutagen toxicity. Cytogenetic analysis of sigma mutants approximately 3 weeks after induction shows an association between the sigma phenotype and readily observable (at the 230-300 band level) chromosomal abnormalities (primarily translocations involving that chromosome 11 carrying the functional TK gene) in 30 of 51 induced mutants studied. Using an early clonal analysis of mutants (approximately 2 weeks after induction) 28 of 30 sigma mutants showed chromosome 11 rearrangements. All lambda mutants studied (17 of 17 evaluated 3 weeks after induction and 8 of 8 evaluated 2 weeks after induction) showed normal karyotypes (at the 230-300 band resolution level), including the chromosome 11s. These observations support the hypothesis that sigma (and likely tau) mutants represent chromosomal mutations and lambda mutants represent less extensive mutations affecting the TK locus. The inclusion of sigma mutants in the total induced mutant frequency, as well as distinguishing them as a separate subpopulation of TK-deficient mutants, is, therefore, essential in obtaining maximum utility of the information provided by the L5178Y/TK+/- mouse lymphoma assay.