Metabolism of tritiated C19 steroids by shionogi mouse mammary tumors

In the present report, the metabolism of dehydroepiandrosterone, 5-androstene-3β, 17β-diol and androstenedione has been studied in two different Shionogi tumors, one is androgendependent (“androgen sensitive”) and grows in intact male mice and the other is apparently androgen-independent (“androgen insensitive”) since the cells continue to grow in castrated antiandrogen (Flutamide) treated male animals. Our data clearly show that both the sensitive and insensitive tumors contain 3β-hydroxysteroid Δ⁵-Δ⁴ isomerase which causes the transformation of C₁₉ steroids into potent androgenic steroids. However, the androgen sensitive tumor is able to convert 5-androstene-3β, l7β-diol into 2-hydroxyestrogens while the rate of conversion is extremely low in the insensitive tumor. Most interestingly, the production of 5α-reduced steroids observed in both tissues was clearly higher in insensitive tumor homogenates.     

Role of the reticulum in the stability and shape of the isolated human erythrocyte membrane

In order to examine the widely held hypothesis that the reticulum of proteins which covers the cytoplamsic surface of the human erythrocyte membrane controls cell stability and shape, we have assessed some of its properties. The reticulum, freed of the bilayer by extraction with Triton X-100, was found to be mechanically stable at physiological ionic strength but physically unstable at low ionic strength. The reticulum broke down after a characteristic lag period which decreased 500-fold between 0 degrees and 37 degrees C. The release of polypeptide band 4.1 from the reticulum preceded that of spectrin and actin, suggesting that band 4.1 might stabilize the ensemble but is not essential to its integrity. The time-course of breakdown was similar for ghosts, the reticulum inside of ghosts, and the isolated reticulum. However, at very low ionic strength, the reticulum was less stable within the ghost than when free; at higher ionic strength, the reverse was true. Over a wide range of conditions the membrane broke down to vesicles just as the reticulum disintegrated, presumably because the bilayer was mechanically stabilized by this network. The volume of both ghosts and naked reticula varied inversely and reversibly with ionic strength. The volume of the naked reticulum varied far more widely than the ghost, suggesting that its deformation was normally limited by the less extensible bilayer. The contour of the isolated reticulum was discoid and often dimpled or indented, as visualized in the fluorescence microscope after labeling of the ghosts with fluoroscein isothiocyanate. Reticula derived from ghosts which had lost the ability to crenate in isotonic saline were shriveled, even though the bilayer was smooth and expanded. Conversly, ghosts crenated by dinitrophenol yielded smooth, expanded reticula. We conclude that the reticulum is a durable, flexible, and elastic network which assumes and stabilizes the contour of the membrane but is not responsible for its crenation.     

Behavioural and electrophysiological responses by reproductive female Neogobius melanostomus to odours released by conspecific males

Behavioural and electrophysiological responses of reproductive and non‐reproductive female round gobies Neogobius melanostomus to water previously occupied by male round gobies (reproductive male water) were compared. Reproductive females spent more time than non‐reproductive females in a tank near the input of water conditioned from reproductive males. Also, reproductive females swam significantly faster than non‐reproductive females, suggesting that reproductive male odour may have activated spawning behaviour. Olfactory epithelial field potential measurement (electro‐olfactogram, EOG) showed that reproductive male water was a potent olfactory stimulus to reproductive females, but not to non‐reproductive females. Reproductive females responded significantly more than non‐reproductive females to solid‐phase (octadecylsilane) extracts of reproductive male water. Also, when these extracts were separated on reverse phase high performance liquid chromatography (HPLC), reproductive females showed noticeably greater responses than non‐reproductive females to the fractions that eluted between 30 and 40 min. The behavioural data support the hypothesis that reproductive male round gobies release compounds into the water that attract potential mates. The EOG data indicate that these compounds can be quantitatively extracted from the water and be partially purified by HPLC. The evidence is not sufficient to indicate whether or not the compounds are steroids. The relatively early elution time on HPLC, however, suggests that if these compounds are steroids, then it is more likely that they will be conjugated rather than free steroids.     

Regeneration of Transgenic Soybean (Glycine max) Plants from Electroporated Protoplasts

Transgenic soybean (Glycine max [L.] Merr.) plants were regenerated from calli derived from protoplasts electroporated with plasmid DNA-carrying genes for a selectable marker, neomycin phosphotransferase (NPTII), under the control of the cauliflower mosaic virus 35-Svedberg unit promoter, linked with a nonselectable mannityl opine synthesis marker. Following electroporation and culture, the protoplast-derived colonies were subjected to kanamycin selection (50 micrograms per milliliter) beginning on day 15 for 6 weeks. Approximately, 370 to 460 resistant colonies were recovered from 1 × 10⁶ electroporated protoplasts, giving an absolute transformation frequency of 3.7 to 4.6 × 10⁻⁴. More than 80% of the kanamycin-resistant colonies showed NPTII activity, and about 90% of these also synthesized opines. This indicates that the linked marker genes were co-introduced and co-expressed at a very high frequency. Plants were regenerated from the transformed cell lines. Southern blot analysis of the transformed callus and leaf DNA demonstrated the integration of both genes. Single-plant assays performed with different plant parts showed that both shoot and root tissues express NPTII activity and accumulate opines. Experiments with NPTII and mannityl opine synthesis marker genes on separate plasmids resulted in a co-expression rate of 66%. These results indicate that electroporation can be used to introduce both linked and unlinked genes into the soybean to produce transformed plants.     

Action of acetazolamide on the chick embryo during late development.

Acetazolamide was injected into chick embryos on the 14th or 15th day of incubation. Doses ranging between 5 and 10 mg per egg produced a retardation in the growth of long bones. The affected bones contained a normal proportion of mineral as determined by ashing and presented a normal histological picture. On the basis of these findings, it is suggested that the alterations were not due to a specific direct effect of the drug on bones. The incorporation of 131-I by the thyroid glands of acetazolamide-injected embryos was analyzed radioautographically and quantitated on the same 6 mu-paraffin sections, with a thin window Geiger counter. The incorporation appeared notably reduced 3 h after the injection of acetazolamide and the reduction persisted for a least 24 h.the electron microscopical observation of thyroid follicular cells from similarly treated embryos showed that the cytological characteristics indicating an active protein synthesis were unmodified with respect to those found in control embryos. These results may indicate that acetazolamide inhibits the iodination of the throid hormone without interfering with the synthesis of the globulin. It is suggested that the growth retardation observed in the embryos treated with acetazolamide may be secondary to the action of the drug on the thyroid gland, although this action appears to be a transitory one.     

Competitive dominance of wild and domestic Norway rats (Rattus norvegicus)

The objective was to determine differences in the relative dominance of wild and domestic Norway rats (Rattus norvegicus) when competing for food in inter-stock, round-robin pairings. Each day, after food-deprivation, wild and domestic rats were individually placed in a testing arena containing a slice of apple. On day 20 and every fourth day thereafter, eight wild-domestic pairs were allowed to compete for the apple. Domestic subjects spent more time eating than did wild rats and were more often dominant. Competition tended to wane over test days in response to defeat in earlier trials. The outcome of competition trials was not changed by increasing the extent of food deprivation together with continuous pairing. A reduction in social inhibitions accompanying the domestication process provides the best explanation for the enhanced competitive potential of the domestic rat.     

L-656,224 (7-chloro-2-[(4-methoxyphenyl)methyl]-3- methyl-5-propyl-4-benzofuranol): a novel, selective, orally active 5-lipoxygenase inhibitor

L-656,224 (7-chloro-2-[(4-methoxyphenyl)methyl]-3-methyl-5-propyl-4-benzofuranol) was a potent inhibitor of leukotriene biosynthesis in intact rat and human leukocytes and CXBG mastocytoma cells (IC50 values, 18-240 nM) and of crude human leukocyte and highly purified porcine leukocyte 5-lipoxygenase (IC50 value, 4 X 10(-7) M). The selectivity of L-656,224 for 5-lipoxygenase was shown through the relative lack of activity of the compound on 12-lipoxygenase, 15-lipoxygenase, cyclooxygenase, catalase, and myeloperoxidase. The compound showed (i) oral activity against hyperalgesia induced in the rat paw by injection of yeast or platelet-activating factor, (ii) dyspnea in sensitized inbred rats induced by an aerosol of antigen, and (iii) bronchoconstriction induced by an aerosol of Ascaris in squirrel monkeys, suggesting a role for 5-lipoxygenase inhibitors in the treatment of asthma and peripheral pain.     

Polarity: the role of localized secretion

Studies of single cells from brown algae suggest that localized secretions stabilize the polar axis resulting in an asymmetry in the cell wall. This cortical asymmetry appears to play a role in orienting the plane of cell division and in determining the different fates of the resulting daughter cells. Recent studies indicate that similar processes may operate in seed plants.