Purification and immunochemical characterization of a low-pI form of UDP glucuronosyltransferase from mouse liver

A liver UDP glucuronosyltransferase (GT) enzyme from either phenobarbital- or 3-methylcholanthrene-treated C57BL/6N mice was isolated by phenyl-Sepharose, DEAE-ion exchange, and UDP hexanolamine chromatographic steps. This enzyme had a broad substrate specificity and was mainly responsible for the microsomal capacity to glucuronidate testosterone, 1-naphthol, and morphine. This UDP glucuronosyltransferase ( GTM1 ) appeared to be at least 95% homogeneous and had a subunit molecular weight of 51,000 using sodium dodecyl sulfate-polyacrylamide gel and two-dimensional gel electrophoreses. Antibodies prepared against the purified protein developed a single immunoprecipitin line by double-diffusion analysis with purified antigen and with solubilized microsomes from both control and drug-induced C57BL/6N and DBA/2N mice. A precipitin line was also observed with microsomal proteins which isoelectrofocused at approximately pH 6.7, but not with those which isoelectrofocused at approximately pH 8.5. GTM1 was, therefore, designated at low-pI form. Immunopurified antibody preferentially inhibited and immunoprecipitated GT activities toward testosterone, 1-naphthol, and morphine. To a lesser extent, activities toward phenolphthalein, 3-hydroxybenzo[a]pyrene, and estrone were inhibited while activities toward 4-nitrophenol and 4-methylumbelliferone were not affected. All activities, however, were immunoadsorbed in the presence of protein A-Sepharose. This observation can be explained by the following results. Immunoprecipitates from labeled microsomes contained primarily a 51,000-Da protein. When the immune complexes were adsorbed with protein A-Sepharose, a 54,000-Da protein as well as the expected 51,000-Da GTM1 was detected. This 54,000-Da protein was associated with the glucuronidation of 3-hydroxybenzo[a]pyrene and 4-nitrophenol, and was designated GTM2 .     

Increase in nuclear cyclic GMP-dependent protein kinase following partial hepatectomy

The level of cyclic GMP-dependent protein kinase in the nucleus of rat liver was shown to increase 80% at 3 hr following partial hepatectomy while cyclic AMP-binding activity decreased 28%. Subnuclear fractionation demonstrated that the increase in cyclic GMP-dependent protein kinase was localized to the nucleolus and nucleoplasm, with no change in the extranucleolar particulate material. Cyclic AMP-binding activity was decreased in all subnuclear fractions under these conditions. At 16 hr following partial hepatectomy, the level of cyclic GMP-dependent protein kinase was not changed in the nucleolus but was significantly increased in the nucleoplasm, while cyclic AMP-binding activity was slightly increased in the nucleolus and decreased in the nucleoplasm.     

Fatal pneumonia with terminal emaciation in nude mice caused by pneumonia virus of mice

Athymic nude mice used as sentinel animals in a mouse holding room died of pneumonia 17 to 32 weeks after being placed in the room. Lesions in the pulmonary parenchyma consisted of monocytic exudate, epithelial cell necrosis, hemorrhage, fibrin deposition and interstitial fibrosis. Septal edema, septal cell necrosis and septal capillary stasis were common, but there was limited sloughing of bronchial lining epithelium. Indirect fluorescence microscopy (IFA) of lung sections using pneumonia virus of mice (PVM) antibody was positive. The pneumonia and IFA results were reproduced in euthymic mice inoculated experimentally with lung suspension from naturally infected mice or with tissue culture fluid from cultures infected with American Type Culture Collection PVM. The lungs of a naturally infected nude mouse were studied by transmission electron microscopy. Virus growth was found on Type II alveolar epithelium and on poorly differentiated replacement alveolar epithelium. Virus particles appeared as long exophytic filaments containing one to six linearly arranged nucleocapsids. Inclusion bodies and intracellular virus structures were not observed.     

Renicola metacercariae (Digenea: Renicolidae) in clupeoid fish: new host records

Two distinct types of Renicola metacercariae were found in herring, Clupea harengus L., from Scottish waters, and one in anchovies, Engraulis encrasicholus (L.), from the central North Sea. Both constitute new host records. These parasites may prove to be of value as biological tags for herring and other clupeoid fish.     

Flow microfluorometric analysis of living spermatozoa stained with Hoechst 33342

Bovine spermatozoa were stained with Hoechst 33342. The fluorescence distribution of stained spermatozoa was complex. Non-motile spermatozoa displayed a higher fluorescence than did motile spermatozoa. The fluorescence profile of the motile spermatozoa was bimodal. Sort and reanalysis, and orientation experiments suggested that there are two distinct populations of motile spermatozoa.     

Simple and stereoselective syntheses of nucleoside analogues with a benzo[c]furan glycone moiety.

A series of d4T analogues have been synthesised in which the 2',3'-didehydro-2',3'-dideoxyribose moiety is replaced by a benzo[c]furan core. A simple strategy has been developed to access a range of compounds for biological screening. In addition, a stereoselective approach has been achieved with view to permit more detailed studies.