Leonard J. Beuving

Androgen-dependency of aggressive target-biting and paired fighting in male mice

Abstract The effect of gonadal hormone manipulations by means of castration and/or exogenous injections on aggressive biting of an inanimate bite-target and wheel running by mice was studied. Castration lowered the bite-attack levels of mice but daily injections of testosterone restored these bite attack levels to that of intact mice. Cessation of the testosterone replacement was again followed by lower bite attack levels. Running activity was monitored to assure specificity of independent variable manipulations and standard paired fighting tests were conducted for comparison with traditional methods. There was a high correlation between the number of paired fight responses and the total number of inanimate target attacks. Running activity did not vary across treatments. Results indicate that androgen is critical for the maintenance of biting attack in males, that androgen affects aggression without altering running, that there is a maximal effect beyond which additional androgen produced little change, and that the single subject paradigm utilized in this study is an effective model for the study of pharmacological and physiological effects on aggressive behavior.

An ultrastructural evaluation of acute 1-nitronaphthalene induced hepatic and pulmonary toxicity in the rat

1-Nitronaphthalene is a mutagenic particulate of diesel exhaust which causes acute liver and lung toxicity in rodents. The studies presented here describe morphological changes in the lung and liver at several time intervals following a single injection of 1-nitronaphthalene (100 mg/kg, i.p.) in male Sprague-Dawley rats using transmission and scanning electron microscopy. Although both the lungs and liver are injured by 1-nitronaphthalene, the lungs appear to be the primary target organ. Within 4 h of treatment, all 1-nitronaphthalene treated animals exhibited respiratory distress characterized by labored breathing, severe gasping and chromodacryorrhea. The primary ultrastructural alteration were hydropic changes in the non-ciliated bronchiolar (Clara) cells of the distal-most bronchioles of the lung. These were apparent as early as 1 h after 1-nitronaphthalene injection, while adjacent ciliated cells showed no alterations. Over a 24 h period, the bronchioles showed progressive ultrastructural changes leading to necrosis and exfoliation of both ciliated and Clara cells. Interstitial pneumonitis and edema were observed in all animals treated with 1-nitronaphthalene, and was usually associated with bronchioles containing necrotic epithelium. In the liver, ultrastructural changes were observed in the centrilobular hepatocytes at 8 h and consisted of cytomegaly, loss of continuous inner membrane and reduced matrix density of the mitochondria. At 48 h, cellular damage to centrilobular hepatocytes was severe and nearly all mitochondria were damaged. Elevated levels of alanine aminotransferase, aspartate aminotransferase and bilirubin were detected in the serum of animals treated with 1-nitronaphthalene at 8-48 h. In conclusion, 1-nitronaphthalene is a pulmonary toxicant with a unique progression of injury, which primarily damages Clara cells followed by ciliated cells. This disparity is likely due to a difference in the bioactivation of 1-nitronaphthalene. Furthermore, this systemic toxicant also has injurious effects on the centrilobular region of the liver which precedes lung injury.

The effects of gormone manipulations on aggressive target-biting in mice

Aggressive biting of an inanimate target by mice was studied. Males attacked the bite-target more frequently than females, but this difference disappeared after castration when the response rate of the males approached that of the females. Ovariectomizing the females had little effect on their bite-attack frequencies. Subsequent androgen injections restored the biting-attack frequency of the castrated males to preoperative levels but had little effect on the intact males. Estrogen had little effect on the response frequency of the females, whereas androgen produced a slight increase in their bite-attack frequency. Results indicate that androgen is critical for the maintenance of this aggressive response and that the single subject paradigm utilized in this study was a sensitive measure of aggressive tendencies in mice.

Sinusoidal endothelial cell and parenchymal cell injury during endotoxemia and hepatic ischemia-reperfusion: protection by the 21-aminosteroid tirilazad mesylate

Abstract The early vascular injury in the liver was characterized in an experimental model of multiple organ failure (MOF). Significant increases of hyaluronic acid levels (660%) and plasma alanine aminotransferase activities (1050%) were observed after 20 min hepatic ischemia followed by 4 h reperfusion and injection of 0.5 mg/kg Salmonella enteritidis endotoxin at 30 min reperfusion. Morphological evaluation of sinusoids with transmission electron microscopy indicated neutrophil and Kupffer cell activation as well as damage or loss of sinusoidal endothelial cells. Hepatocellular injury was evident from fused microvilli and blebbed plasma membranes. Treatment with the 21-aminosteroid tirilazad mesylate (U-74006F) (2 × 3 mg/kg) reduced plasma hyaluronic acid levels by 61% and plasma transaminase activities by 69% suggesting a beneficial effect on sinusoidal endothelial cell and parenchymal cell injury. This was confirmed by morphology. Our data provide morphological and functional evidence for severe injury to sinusoidal endothelium and the vascular pole of hepatocytes in this model of MOF. U-74006F significantly protected the liver against this Kupffer cell- and neutrophil-mediated injury. Thus, U-74006F may be a promising therapeutic for liver dysfunction and failure during a local or systemic inflammatory response.

The Effects of Insulin on Hepatic Glucocorticoid Receptor Content in the Diabetic Rat

Streptozotocin-induced diabetic rat liver was analyzed for glucocorticoid receptor (GR) content by saturation and Scatchard analysis. The hepatic GR content of streptozotocin-induced diabetic rats was significantly decreased from a control level of 0.17 +/- .01 pmol/mg protein to 0.11 +/- .01 pmol/mg protein. Insulin replacement therapy to the diabetic rat dramatically increased the hepatic GR content to 0.26 +/- 0.02 pmol/mg protein as compared to the diabetic value of 0.11 +/- 0.01 pmol/mg protein. A time course study of GR content in the diabetic rat liver demonstrated that after an initial decrease in hepatic GR content at 14 days, the 25-day diabetic receptor level elevated back to control levels. A significant increase in GR content over controls was observed in the 110-day diabetic rats. These results suggest that insulin has a role in the regulation of hepatic GR content.

A Photoreactive Fluorescent Marker for Identifying Eosinophils and Their Cytoplasmic Granules in Tissues

Here we describe a simple histochemical technique that provides an improved approach to identifying eosinophil components in tissues through the formation of photoreactive complexes that produce stable fluorescent emissions. This method worked readily with histological tissue sections 6–60 μm thick, which were fixed in neutral buffered formalin (NBF), and with cell suspensions similarly fixed and unfixed. Deep red (>605 nm) fluorescent emissions were produced by eosinophil-specific granules when exposed to broadband excitation spectra from a 100-W mercury lamp source (510–590 nm), as well as single-wavelength excitations from both an argon laser (488 nm) and a UV-visible laser (514 nm). The fluorophore-granule complex emissions increased in intensity during the first minute of continuous photoexcitation, then remained stable (>10 min). All nonspecific autofluorescence phenomena associated with these tissues were photobleached in the first minute, including areas of background Biebrich scarlet binding where photoreactive complexes were not formed (i.e., collagen), indicating environmental influences on the fluorophore. This technique allows the visualization of eosinophil granules over a greater period of time than is usually permissible with standard fluorescent markers. Therefore, techniques such as confocal microscopy can be utilized to their fullest extent, providing much more detailed information on the location and distribution of the cytoplasmic contents of eosinophils.