Phosphorylation of the androgen receptor by a nuclear cAMP-independent protein kinase

The androgen receptor was purified from rat ventral prostate. The purified receptor migrated as a single band of mol. wt. 87000 on SDS-polyacrylamide gels, had a kd for R-1881 (17 beta-hydroxy-17 alpha-methyl-estra-4,9,11-trien-3-one) binding as 6 nM, and sedimentation coefficient of 4.5 S. Phosphorylation of the purified receptor was studied by incubating it with [gamma-32P]ATP in the presence of several purified protein kinases including cAMP-dependent protein kinase, and four cAMP-independent protein kinases (which were active towards substrates such as phosvitin and casein). Phosphorylation of the 87000 mol. wt. androgen receptor protein occurred only in the presence of a nuclear cAMP-independent protein kinase (of the N2 type). No auto-phosphorylation of the receptor was detected. The results indicate that the androgen receptor is a phosphoprotein. Further, phosphorylation of the androgen receptor by only a specific nuclear cAMP-independent protein kinase may be important in determining the dynamics of its function.     

Role of glutathione in the regulation of hepatic cholesterol 7 alpha-hydroxylase, the rate-limiting enzyme of bile acid biosynthesis

The effect of in vivo variation of hepatic glutathione (using diethyl maleate and L-cysteine) on in vitro cholesterol 7 alpha-hydroxylase activity was studied in male Sprague-Dawley rats. Cholesterol 7 alpha-hydroxylase activity in glutathione-depleted rats (ca. 10% of control glutathione) was significantly reduced compared to that in vehicle-injected controls. While L-cysteine treatment of glutathione-depleted animals increased glutathione levels somewhat (ca. 20% of control glutathione), they were still significantly less than control levels. Similarly, cholesterol 7 alpha-hydroxylase activity in the partially glutathione replete animals was approximately 50% greater than that in the glutathione-depleted animals, but still significantly less than that in the controls. The rate of 7 alpha-hydroxylation of cholesterol was found to be dependent on liver glutathione content. The calculated maximal rate was 34.4 picomoles/mg/min with a half maximal activity at 1.89 mumoles glutathione/gm liver. These results suggest that hepatic glutathione may be an important modulator of in vivo activity of cholesterol 7 alpha-hydroxylase.     

Effects of polyamines on prostatic chromatin- and non-histone-protein-associated protein kinase reactions.

Studies are presented on the influence of polyamines on prostatic chromatin- and non-histone-protein-associated protein kinase reactions involving both exogenous and endogenous substrates. The activities toward the model acidic protein substrate, dephosphophosvitin, were maximal at 160--200mM-NaCl (or -KCl or -NH4Cl). Under these conditions, spermidine and spermine added in concentrations up to 2mM were essentially without effect. However, without addition of NaCl to the medium, marked stimulation of these reactions was elicited by these polyamines at 1--2mM concentrations. The stimulatory effects were not due to non-specific changes in the ionic strength or to substitution of spermine for Mg2+, as maximal stimulation by 1 mM-spermine was observed only at optimal (2--4mM) Mg2+ concentrations. Qualitatively similar effects of polyamines were observed with enzyme preparations from the prostates of castrated rats, and with chromatin and non-histone-protein preparations from other tissues besides ventral prostate. When phosphorylation of endogenous non-histone proteins of the chromatin was measured, spermine stimulated both the initial rates and the final extent of transphosphorylation, even in the presence of optimal concentration of NaCl. By contrast, spermine or spermidine had no effect on the chromatin- and non-histone-protein-associated protein kinase reactions determined with lysine-rich histones as substrates. Chemically NN-dimethylated dephosphophosvitin was a less active substrate for the chromatin-associated protein kinase, but its phosphorylation was more markedly stimulated by spermine in comparison with unmodified dephosphophosvitin. These observations hint that the polyamine stimulations of the various protein kinase reactions may be due to effects on the conformations of the non-histone protein substrates rather than on the kinases themselves.     

Der Einfluss des Wirtes in der Massenzucht auf die Qualität des EiparasitenTrichogramma evanescens bei der Bekämpfung des Maiszünslers,Ostrinia nubilalis

Zusammenfassung In vier Feldversuchen wurde die Wirksamkeit eines an den Maiszünsler besonders angepassten Stammes vonTrichogramma evanescens Westw. bei der Bek?mpfung des MaiszünslersOstrinia nubilalis Hübner geprüft. Die Parasiten wurden entweder auf Eiern der GetreidemotteSitotroga cerealella (Oliv.) oder auf Eiern der MehlmotteEphestia kuehniella Zell. gezüchtet. Die Freilassung erfolgte in den Versuchsparzellen durch Anbringen von Eikarten mit jeweils etwa 1000 parasitierten Wirtseiern an Bl?ttern von Maispflanzen in Abst?nden von 14 Metern. VierTrichogramma-Freilassungen in w?chentlichen Abst?nden mit jeweils 45000 Parasiten pro Hektar ergaben hohe Parasitierungsraten bei denO. nubilalis-Eiern und starke Verminderungen des Larvenbefalls. In den Parzellen, in denen aufS. cerealella-Eiern gezüchtete Parasiten freigelassen wurden, lagen die Parasitierungsraten zwischen 81,7 und 82,3% im Vergleich zu 63,0 und 84,3% bei den aufE. kuehniella gezüchteten Trichogrammen. Die Verminderung des bei der Ernte festgestelltenO. nubilalis-Larvenbefalls im Vergleich zur unbehandelten Kontrolle lag bei den aufS. cerealella gezüchteten Parasiten zwischen 75,5 und 94,8% verglichen mit 77,6 und 89,9% bei den aufE. kuehniella gezüchteten Tieren. Es gab somit keine signifikanten Unterschiede zwischen den beiden getestetenTrichogramma-St?mmen.

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Summary The effectiveness of a strain ofTrichogramma evanescens Westw. especially adapted to the European corn borerOstrinia nubilalis Hübner was examined in 4 field experiments. The parasites were reared on either the Angoumois grain mothSitotroga cerealella (Oliv.) or the Mediterranean flour mothEphestia kuehniella Zell. and were released in separate experimental plots by attaching egg cards, each containing about 1000 parasitized host eggs, to corn leaves at distances of 14 meters in the field. FourTrichogramma treatments at 1 week intervals using 45000 parasites per hectar per treatment resulted in high parasitization rates ofO. nubilalis-eggs and large reductions in larval damage. In experimental plots whereT. evanescens, reared onS. cerealella-eggs, were released, the parasitization rates ofO. nubilalis-eggs varied between 81,7 and 82,3%, compared with 63,0 and 84,3% forT. evanescens reared onE. kuehniella. The reduction inO. nubilalis larval infestation at the harvest in the treated plots, compared to untreated controls varied between 75,5 and 94,8% forT. evanescens that was reared onS. cerealella, compared with 77,6 and 89,8% forT. evanescens that was reared onE. kuehniella. There were no significant differences in efficiency between the two tested methods of rearingTrichogramma.

     

Microwaves induce an increase in the frequency of complement receptor-bearing lymphoid spleen cells in mice.

A single 30-min exposure of mice to 2450 MHz microwaves (12 to 15 mW/g body weight) in an environmentally controlled waveguide facility induced a significant increase in the proportion of complement-receptor positive lymphoid cells in the spleen. This effect was further enhanced by repeated (three times) exposures, which in addition produced a significant increase in the proportion of Ig+ cells. The proportion of theta-positive cells and the total number of spleen cells remained unchanged.     

Schizogony in Haemoproteus columbae Kruse.

To fill in some of the gaps in our knowledge of Schizogony of Haemoproteus columbae Kruse, transmission experiments involving inoculation into pigeons (Columba livia Gmelin) of sporozonites from salivary glands of the hippoboscid fly Pseudolynchia canariensis (Macquart) were carried out. We were unable to detect prepatent schizonts or to observe schizogonic development when infection became chronic. Schizonts were mainly confined to lung tissue. Observations of parapatent schizonts were made in smears and tissue sections. A variety of forms was found. Cytomeres were rarely encountered. Two types of morphologically distinct merozoites were seen. One type was twice as large as large as the other and was thought to repeat the process of schizogony several times before invading erythrocytes. Schizonts with cytoplasmic clefts were not common in our material due to the fixatives used (Bouin's and Carnoy's). Merozoites were occasionally observed inside monocytes, probably being phagocytosed.