Quantitative determination of the induction of apoptosis in a murine B cell line using flow cytometric bivariate cell cycle analysis.

WEHI-231 cells have been used extensively as a model of tolerance induction in B cells. Recent evidence has shown that anti-IgM treatment of WEHI-231 cells resulted in the induction of apoptosis. In this study, using acridine orange staining and flow cytometric analysis, we demonstrated that apoptotic cells are detected as a distinct population of cells separate from the cells in normal cell cycle progression. The validity of analysis gates was confirmed by cell sorting of the apoptotic population versus normal cells and subsequent gel analysis. Using this technique, we have demonstrated that F(ab')2 anti-mu, A23187, or PMA induced apoptosis in the WEHI-231 cells. The addition of LPS reversed apoptotic induction as seen previously with the WEHI-231 cell line. In contrast, however, PMA did not prevent the induction of apoptosis in anti-mu-treated cells. Additionally, we were interested in determining if the induction of apoptosis was restricted to a specific phase of cell cycle. Since growth inhibition results in most cells arresting in the G1 phase of cell cycle, we wanted to demonstrate apoptosis as a G1-dependent event. This was examined with WEHI-231 cells treated with known cell cycle inhibitors. Interestingly, inhibition of cells in each phase of cycle resulted in the induction of apoptosis. LPS was able to inhibit the induction of apoptosis with each of the cell cycle inhibitors except actinomycin D. Furthermore, we have demonstrated that the WEHI-231 cells contain a Ca(2+)-Mg(2+)-dependent preexisting endonuclease.     

The impact of tick parasites on the behaviour of the lizard Tiliqua rugosa

Populations of the Australian sleepy lizard, Tiliqua rugosa, near Mt. Mary, South Australia carry natural infestations of two tick species Aponomma hydrosauri and Amblyomma limbatum. In field experiments at two sites, 18 km apart, lizards with experimentally increased tick loads had smaller home ranges, moved shorter distances in a day, and were found basking more but moving less often than lizards from which ticks were experimentally removed. The results were consistent for adult lizards in two years, and for sub-adults in a third year. Laboratory trials showed that juvenile lizards that had tick infestations had lower sprint speeds than uninfested siblings, and that adults with tick infestations had less endurance than those that were uninfested. The results contrast with those of a previous survey that showed that lizards with high tick loads had greater body size and remained longer at a site, but indicate that there may be a balance, for lizards, between the fitness advantages in occupying habitats with high-quality resources, and the costs from parasites that also prefer those habitats. Received: 02 March 1999 / Accepted: 07 October 1999     

Elasto-thixotropic properties of bronchial mucus and polymer analogs. I. Experimental results

The non-newtonian viscous and elasto-thixotropic properties of native and lyophilized pathological bronchial mucus and of polymer solutions (3% and 6% PIB in decalin) used as mucus analogs were analyzed using a cone-plate Carri-Med rheometer and a Couette viscoelastometer that we have specifically developed for measuring the rheological properties of bronchial mucus in clinical practice. The master curves obtained for apparent viscosity under steady conditions as a function of shear rates (gamma: 2.6 X 10(-3) to 6.9 X 10(1) sec-1) were fairly similar, whatever the apparatus used. Under transient conditions, at low shear rate (gamma less than 1.4 sec-1), PIB and mucus exhibited a typical viscoelastic behavior: the shear stress increased slightly up to a steady-state value. At higher gamma, a transitory overshoot of sigma characteristic of the elastothixotropic systems appeared. Such a behavior can be interpreted as resulting from structural changes such as formation and rupture of the three-dimensional network present in bronchial mucus as in polymer solutions.     

Sexual behavior and serum hormone levels in aging rhesus males: effects of environmental change

Old (N = 9) and middle-aged (N = 9) rhesus males (Macaca mulatta) that were housed in the same type of home cage and the same room for several years were moved to a new location in an adjacent building. Their sexual behavior and hormone levels in the weeks preceding the move were compared to those after the move. The hypothesis that the nonspecific sensory stimulation provided by the move would increase levels of sexual behavior was not supported by the data. To the contrary, the percentages of tests with contacts, mounts, and penile erections, and the rate of contacting the female, decreased significantly for both old and middle-aged males. The intromission rate of middle-aged, but not of old, males also decreased. In consonance with previous findings, middle-aged males displayed significantly higher levels of sexual activity than did old males. The differences were observed both before and after the move. Serum testosterone levels did not differ between middle-aged and old males, and the move did not produce significant changes in testosterone levels. Serum cortisol levels were significantly higher immediately after the move (1 hr) but returned to premove levels the following day.     

An improved purification method for cytosolic malate dehydrogenase from several sources

A new purification method for cytosolic malate dehydrogenases from several sources has been developed. The procedure, employing chromatographies on 5'AMP-Sepharose, DEAE-Sephacel and Blue-Sepharose, allows for a rapid isolation of the enzyme (approximately 40 hours), in large quantities, with good yields (45-54%). The specific activity of final preparations were around 1300 I.U./mg and were judged homogeneous by polyacrylamide gradient gel and sodium dodecyl sulfate polyacrylamide gel electrophoresis, high performance size exclusion chromatography and isoelectric focusing.