High affinity specific antiestrogen binding sites are concentrated in rough microsomal membranes of rat liver

Saturation and competitive binding analyses demonstrated the presence of a high affinity (K_D = 0.92 nM), specific antiestrogen binding site (AEBS) in rat liver microsomes and at least 75% of total liver AEBS was recovered in this fraction. When microsomes were further separated into smooth and rough fractions, AEBS was concentrated in the latter. Subsequent dissociation of ribosomes from the rough membranes revealed that AEBS was associated with the membrane and not the ribosomal fraction. Antiestrogen binding activity could not be extracted from membranes with 1 M KCl or 0.5 M acetic acid but could be solubilized with sodium cholate. These data indicate that AEBS is an integral membrane component of the rough microsomal fraction of rat liver.     

Computational Investigations of the Chalcogen-Substituted Carboxylic Acids RC(=O)XH and RC(=X)OH and their Dimers [RC(=O)XH]2 and [RC(=X)OH]2 (X=S, Se, Te)

Semiempirical and ab initio theoretical methods have been used to investigate molecular structures of the chalcogen-substituted carboxylic acid isomers RC(=O)XH (chalcogenol acid) and RC(=X)OH (chalcogenon acid). A recent experimental report suggests that the chalcogenon isomers, although less stable at room temperature, predominate at low temperature in polar solvents and that there is only a small barrier to isomerization between the isomers. Theoretical calculations have been used to locate minimum energy structures of chalcogen-substituted carboxylic acid isomers and to calculate energy differences between pairs of isomers. Carboxylic acids are well known to dimerize, especially in the gas phase and in non-polar solvents. We have, therefore, also calculated energies of dimerization of the chalcogen-substituted acids by optimizing the geometries of the symmetric dimers. We note that the PM3 level of theory is only qualitatively correct for sulfur- and selenium-containing species but fails even qualitatively for the tellurium-containing compounds. Ab initio results confirm the experimental observations and provide good estimates of both isomerization and dimerization energies. We conclude that for many functional groups with tautomers RC(=X)YH and RC(=Y)XH, the more acidic tautomer is the one with the acid proton on the smaller, more electronegative atom, although in many cases this may not be the more stable tautomer.Electronic Supplementary Material available.     

Lens regeneration from cultured newt irises stimulated by retina-derived growth factors (EDGFs)

It has been shown that lens regeneration from the iris of the newt Notophthalmus viridescens is dependent on the presence of neural retinal tissue in organ culture and in vivo. The recent discovery of various eye-derived growth factors (EDGFs) in the bovine retina [14] prompted us to investigate whether one of these factors may be involved in the stimulation of lens regeneration. Dorsal irises were cultured for 20 days in serum-supplemented diluted Eagle's medium. Growth factors from bovine retina of various degrees of purification were added. Lens regeneration was assessed on the basis of morphological lens-regeneration stages and by immunofluorescent detection of a lens-specific marker protein, alpha-crystallin. Crude isotonic retinal extract at 80-800 micrograms/ml significantly augmented lens regeneration. Very similar results were obtained when EDGF III, the nonretained retinal factor after heparin-affinity chromatography, was present at 2-20 micrograms/ml. Lens regeneration was also significantly increased when EDGF II, the retinal form of acidic fibroblast growth factor (aFGF) at 50-500 ng/ml was added to the cultures. On the other hand, EDGF I at 4-40 ng/ml and brain basic FGF at 5-50 ng/ml did not seem to significantly stimulate lens regeneration under the conditions used. Our results suggest that at least two retina-derived growth factors (EDGF II and III) can stimulate lens regeneration. These growth factors may be the putative signal that is naturally produced by the retina during lens regeneration in the newt.     

Use of Metabolic Inhibitors to Estimate Protozooplankton Grazing and Bacterial Production in a Monomictic Eutrophic Lake with an Anaerobic Hypolimnion

Inhibitors of eucaryotes (cycloheximide and amphotericin B) and procaryotes (penicillin and chloramphenicol) were used to estimate bacterivory and bacterial production in a eutrophic lake. Bacterial production appeared to be slightly greater than protozoan grazing in the aerobic waters of Lake Oglethorpe. Use of penicillin and cycloheximide yielded inconsistent results in anaerobic water and in aerobic water when bacterial production was low. Production measured by inhibiting eucaryotes with cycloheximide did not always agree with [³H]thymidine estimates or differential filtration methods. Laboratory experiments showed that several common freshwater protozoans continued to swim and ingest bacterium-size latex beads in the presence of the eucaryote inhibitor. Penicillin also affected grazing rates of some ciliates. We recommend that caution and a corroborating method be used when estimating ecologically important parameters with specific inhibitors.