Pharmacological studies on the potentiation of phenytoin teratogenicity by acetaminophen

An in vivo murine model was developed to measure maternal phenytoin biotransformation along with the covalent binding of phenytoin to fetal tissues in the same fetuses which were assessed for fetal anomalies. Acetaminophen was administered to pregnant CD-1 mice 1 hour prior to phenytoin, both given i.p. at varying doses and gestational times between days 11 and 13. Dams were killed between days 12 and 19. Metabolites reflecting the enzymatic bioactivation of phenytoin were quantified in maternal plasma and urine with high-performance liquid chromatography (HPLC). Acetaminophen pretreatment caused a threefold increase in phenytoin-induced fetal cleft palates without increasing resorptions. The covalent binding of radiolabeled phenytoin to fetal and placental tissues measured on day 13 was increased twofold and threefold, respectively, by acetaminophen pretreatment. Phenytoin covalent binding measured on day 16 was significantly increased in the livers of fetuses with cleft palates, but not in the livers of dams with fetuses having cleft palates. Binding to fetal brain on day 16 was over fourfold higher than that in maternal brain. Acetaminophen pretreatment differentiated dams into poor and extensive metabolisers of phenytoin, with only the latter group carrying fetuses with cleft palates. The incidence of fetal cleft palates correlated positively with maternal urinary levels of phenytoin (r = +.81, P less than .01) and its dihydrodiol metabolite (r = +.61, 0.05 less than P less than .1), and negatively with levels of para-hydroxylated phenytoin (r = -.85, P less than .01). These findings related both to the mechanism of phenytoin teratogenicity and its potentiation by acetaminophen.     

Tryptophan auxotrophs of Rhizobium japonicum

Eleven tryptophan-requiring mutants of Rhizobium japonicum I-110 ARS were isolated after nitrous acid mutagenesis and fell into five groups based on characterization by supplementation with intermediates and enzyme assays.     

Detection of gonadotrophic hormones on isolated granulosa cells of the domestic hen, Gallus domesticus, by an immunohistochemical method

Two immunohistochemical methods were used to detect the presence of luteinizing hormone (LH) on the cells of chicken granulosa. Using a peroxidase-labelled anti-rabbit serum together with anti-chicken LH serum raised in rabbits, a strong positive response was obtained with granulosa cells from small and large pre-ovulatory follicles obtained from the mid-cycle. Similarly, by using an available antiserum to human follicle stimulating hormone (FSH), a slightly weaker response was obtained with cells from both large and small follicles. After incubating cells with ovine LH, ovine FSH and ovine prolactin, there was no detectable difference with the method used in reaction with their respective antisera to cells which had received no incubation, implying that chicken gonadotrophins were displaced only partially from their receptors by mammalian gonadotrophic hormones. Pre-incubation of the cells with human chorionic gonadotrophin gave negative results with anti-hCG serum. Using a fluorescent-labelled antibody method, similar results were obtained except that the distribution of the receptors on the granulosa cell for LH or FSH appeared to be different. With the LH, the fluorescence formed a halo around the cell in contrast to the overall fluorescence with FSH.     

Population equilibria and stability in plant-animal pollination systems

Differential equation models of the change in population size with change in time have provided valuable insight into nature and have served as tools for management of both predator-prey and competition systems. Presented here is an extension of the differential equation models for application to pollination ecology plant-animal interactions. Analyses define conditions for equilibria and for equilibrium stability for a one-plant one-animal pollinator system. Predictions about plant speciation and pollinator foraging behavior are discussed.     

Cloned nodulation genes of Rhizobium leguminosarum determine host-range specificity

Summary Three nodulation-deficient (nod) mutants of Rhizobium leguminosarum were isolated following insertion of the transposon Tn5 into pRL1JI, the R. leguminosarum plasmid known to carry the nodulation genes. DNA adjacent to the nod: Tn5 alleles was subcloned and used to probe a cosmid clone bank containing DNA from a Rhizobium strain carrying pRL1JI. Two cosmid clones which showed homology with the probe contained about 10 kb of DNA in common. The R. leguminosarum host-range determinants were found to be present within this 10 kb common region since either of the cosmid clones could enable a cured R. phaseoli strain to nodulate peas instead of Phaseolus beans, its normal host. Electron microscopy of nodules induced by Rhizobium strains cured of their normal symbiotic plasmid but containing either of the two cosmid clones showed bacteroid-forms surrounded by a peri-bacteroid membrane, indicating that normal infection had occurred. Thus it is clear that this 10 kb region of nodDNA carries the genes that determine host range and that relatively few bacterial genes may be involved in nodule and bacteroid development.     

Recovery from a viral respiratory tract infection. IV. Specificity of protection by cytotoxic T lymphocytes

Immune spleen cells enhanced for influenza-specific cytotoxic activity after exposure to virus-infected stimulator cells in vitro effect recovery when transferred to nude and immunocompetent mice with influenza pneumonia (5). This protective effect correlated with the virus-specific cytotoxic activity of the transferred lymphocytes and is removed by treatment with anti-0 serum and complement. The experiments presented here indicate that spleen cells taken directly from mice undergoing a primary or secondary infection are less protective than immune spleen cells that are restimulated in vitro before transfer. This decreased ability to clear pulmonary virus and effect survival correlated with their relatively lower levels of influenza-specific cytotoxicity. Protection did not correlate with the level of natural killer cell activity of transferred cells. The results also indicate the immune spleen cells that are protective are influenza A subtype cross-reactive and are H-2-restricted; H-2d immune spleen cells effected recovery of H-2d but not H-2k challenged mice.     

New Method of Isolating Salmonellae from Milk

The use of a cotton gauze swab and subsequent culture of the swab was found to be a more sensitive method for isolating Salmonella from liquid milk than the revised procedure of North. The swab method was found to be as sensitive as the North procedure for recovering Salmonella when incubated at 37 C but more sensitive when incubated at 43 C. Incubation of the swab cultures at the elevated temperature of 43 C gave good results when Salmonella was present at levels as low as one per liter. Swabs exposed to milk contaminated with 100 Salmonella per liter remained positive even when subsequently washed for 2 hr in noncontaminated milk. Bismuth sulfite agar and Brilliant Green sulfadiazine agar were equally effective for isolating Salmonella from broth cultures; use of both media resulted in maximal isolations.