Structure of the precursor to an enzyme mediating COOH-terminal amidation in peptide biosynthesis

Many bioactive peptides terminate with an amino acid alpha-amide at their COOH terminus. The enzyme responsible for this essential posttranslational modification is known as peptidyl-glycine alpha-amidating monooxygenase or PAM. We identified cDNAs encoding the enzyme by using antibodies to screen a bovine intermediate pituitary lambda gt11 expression library. Antibodies to a beta-galactosidase/PAM fusion protein removed PAM activity from bovine pituitary homogenates. The 108,207 dalton protein predicted by the complete cDNA is approximately twice the size of purified PAM. An NH2-terminal signal sequence and short propeptide precede the NH2 terminus of purified PAM. The sequences of several PAM cyanogen bromide peptides were localized in the NH2-terminal half of the predicted protein. The cDNA encodes an additional 430 amino acid intragranular domain followed by a putative membrane spanning domain and a hydrophilic cytoplasmic domain. The forms of PAM purified from bovine neurointermediate pituitary may be generated by endoproteolytic cleavage at a subset of the 10 pairs of basic amino acids in the precursor. High levels of PAM mRNA were found in bovine pituitary and cerebral cortex. In corticotropic tumor cells, levels of PAM mRNA and pro-ACTH/endorphin mRNA were regulated in parallel by glucocorticoids and CRF.     

Ketone-body metabolism after partial hepatectomy in the rat.

Fed or 24 h-starved rats were subjected to two-thirds partial hepatectomy or sham-operation and subsequently starved for 4, 14 or 24 h. Despite high plasma fatty acid concentrations, the partially hepatectomized rats failed to respond to post-operative starvation with increased blood and liver ketone-body concentrations or to maintain the high ketone-body concentrations associated with pre-operative starvation. Hypoglycaemia and hyperlactaemia were observed within 30 min of functional hepatectomy, but not partial hepatectomy, of 24 h-starved rats, and, even after a further 24 h starvation of partially hepatectomized rats, blood glucose concentrations were only slightly decreased. The results are discussed with reference to fat oxidation and gluconeogenesis in the liver remaining after partial hepatectomy.     

The persistence of Legionella pneumophila in non-sterile, sterile and artificial hard waters and their growth pattern on tap washer fittings

Simultaneous experiments were performed with sterilized and non-sterile water and an artificial hard water. After seeding with an environmental isolate of Legionella pneumophila numbers in the sterile land hard water decreased rapidly and colonization of various tap washer fittings failed to take place. Adhesion and growth of an environmental isolate of L. pneumophila to washers in non-sterile tap water was followed over a 4-month period with fluorescein-labelled antibody and by scanning electron microscopy. After adherence the individual cells appeared to divide to form chains which spread over the surfaces. Organisms other than legionellas were also present and a complex colonization matt was formed which was embedded in a protective coat of slime and debris. The numbers of L. pneumophila recovered from the water were highest between 4 and 7 weeks but they could still be cultivated after 4 months.     

Industrial contributions to desertification in South America

Degradation of tropical lands has a grim inevitability. This is true of semi-arid and seasonal ecosystems as it is of the rainforests, whose plight has been the principal focus of attention in recent years. The broad belt of savanna-like vegetation in central South America has suffered an accelerating decline in productivity due to overclearing and overgrazing, a process that can be traced directly to the industrialization of Europe in the 19th century. The remorseless slide towards unproductivity need not be slow: a mere 50-100 years seems more than adequate, even for the more robust landscapes. With careful management however, some of these lands are slowly recovering.