Oxygen-derived free radical inhibition in the healing of experimental zone-of-stasis burns.

When dehydration, infection, and mechanical trauma are prevented, procedures (such as cooling and/or oral antithromboxane) designed to diminish ischemia in experimental zone-of-stasis burns have been associated with no or only minor improvement in wound healing. To test the hypothesis that ongoing skin damage occurring postburn (PB) may in part be due to release of oxygen-derived free radicals during the 16-hour through 4-day PB period of reperfusion in such burns, beginning immediately and for a period of 5 days PB, equal numbers of guinea pigs received: allopurinol 150 mg/kg PO q 6 h vs. placebo, dimethylsulfoxide (DMSO) 75% applied topically q 12 h vs. placebo, or yeast-derived superoxide dismutase coupled with polyethylene glycol (PEG-SOD, Pharmacia) 10,000 U (Fridovich) given IV q 8 h producing a concentration of 16 U/cc of plasma 8 hr after injection vs. placebo. Gross and histologic examination of wounds by a 'blinded' investigator at 1 week and 3 weeks PB revealed no difference between treatment and control groups when rates of re-epithelialization and frequencies of hair-follicle retention were compared. Using the dosages, routes, and model described, treatment of a zone-of-stasis burn with PO allopurinol (a xanthine oxidase inhibitor), topical DMSO (a scavenger of the hydroxyl radical), or IV PEG-SOD (a scavenger of the superoxide radical) during the first 5 days PB was associated with no increase in the rate of re-epithelialization or frequency of hair follicle retention at 1 and 3 weeks PB when compared with controls.

Stromal and nuclear markers for rapid identification of pituitary adenomas at biopsy.

We tested fluorescent and light microscopic markers to improve recognition of pituitary adenomas at biopsy. The optimal reagent was 100 mg/L of fluoresceinated Ricinus communis agglutinin 120 (RCA 120) lectin plus 3 mg/L of propidium iodide. The refrigerated solution was immediately available for use on routine frozen sections. The sections were stained for one minute and viewed immediately after they were rinsed with saline and coverslips applied. Fluorescein-labeled RCA 120, which binds galactose, localized vascular stroma. Propidium iodide, which binds nucleic acids, stained nuclei. Stromal configuration, nuclear morphology, and cell to stroma ratio were illuminated and used to distinguish adenoma from adenohypophysis. We also describe a method utilizing peroxidase-conjugated RCA 120 that demonstrates the same features by light microscopy. Fluoresceinated RCA 120-stained vessels and stroma of routinely processed material more reliably than hematoxylin-eosin or labeled antibody to fibronectin and faster than peroxidase-conjugated RCA 120 or the rapid method for reticulin.

Detection of triethylenemelamine-induced translocation heterozygotes in CD-1 mice: fertility and cytological methods.

A study was conducted to determine the extent to which translocation carriers can be missed in the heritable translocation test. CD-1 male mice were treated with the mutagen triethylenemelamine and bred to untreated females and male progeny were subjected to fertility and cytogenetic analyses. Fertility testing involved mating 1 male to 3 virgin females and subjecting the females to uterine analysis. The males were classified as having normal or reduced fertility according to the number of live implants produced by the females. The cytogenetic analysis involved scoring 25 primary spermatocytes per mouse for translocation multivalents. A total of 103 male progeny were analyzed. Evaluation of fertility and cytogenetic data confirmed the presence of 18 translocation heterozygotes among the male progeny. On the basis of fertility testing, 1 translocation heterozygote was classified as normal (false negative) based on the production of 11 or more live implants by at least 1 mated female. On the basis of cytogenetic analysis of 25 cells per mouse, 1 partially sterile male was classified as normal (false negative); however, analysis of additional cells showed that this mouse was a translocation heterozygote. The study demonstrates the importance of evaluating fertility and cytogenetic criteria to minimize the extent of misclassification in conducting a heritable translocation test.

Biosynthesized products of cultured neuroglial cells: I. Selective release of proteins by cells from human astrocytomas.

The proteins synthesized and released by human astrocytoma cells cultured with radiolabeled amino acids were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate and by trichloracetic acid precipitation. A select group of extracellular proteins was released from the astrocytomas. The most abundant extracellular proteins were at least 250,000 daltons. Five other major proteins (P 175, P 125, P 60, P 47, and P 40) were 175,000, 125,000, 60,000, 47,000, and 40,000 daltons, respectively. The complement of proteins retained by the cells was considerably more complex than those released. Clinical efforts to enhance the immunologic response directed against human astrocytomas must distinguish between extracellular and retained antigens. The pattern of proteins released by gliomas might be diagnostically useful if present in cerebrospinal fluid or serum.