Isolation and characterization of the human pulmonary surfactant apoprotein gene

Pulmonary surfactant is a phospholipid-protein complex which serves to lower the surface tension at the air-liquid interface in the alveoli of the mammalian lung and is essential for normal respiration. Inadequate levels of surfactant at birth, a frequent situation in premature infants, results in respiratory failure. In all species examined, surfactant is composed primarily of dipalmitoylphosphatidylcholine and two major protein species of relative molecular mass (Mr) 32,000 (32K) and 10K (refs 2-5). Reconstitution in vitro of purified 32K pulmonary surfactant apoprotein (PSAP) with synthetic lipids forms a lipoprotein complex that lowers surface tension by spreading to create a thin interfacial film. Here we describe the cloning of the human PSAP gene and complementary DNA, and discuss features of the unusual encoded protein.     

Resistance to beta-lactam antibiotics by re-modelling the active site of an E. coli penicillin-binding protein

The beta-lactam antibiotics kill bacteria by inhibiting a set of penicillin-binding proteins (PBPs) that catalyse the final stages of peptidoglycan synthesis. In some bacteria the development of intrinsic resistance to beta-lactam antibiotics by the reduction in the affinity of PBPs causes serious clinical problems. The introduction of beta-lactam antibiotics that are resistant to hydrolysis by beta-lactamases may also result in the emergence of intrinsic resistance among the Enterobacteriaceae. The clinical problems that would arise from the emergence of resistant PBPs in enterobacteria have led us to examine the ease with which Escherichia coli can gain resistance to beta-lactams by the production of altered PBPs. The development of resistant PBPs also provides an interesting example of enzyme evolution, since it requires a subtle re-modeling of the enzyme active centre so that it retains affinity for its peptide substrate but excludes the structurally analogous beta-lactam antibiotics. We show here that only four amino-acid substitutions need to be introduced into PBP 3 of E. coli to produce a strain possessing substantial levels of resistance to a wide variety of cephalosporins. We also show that transfer of the gene encoding the resistant PBP 3 from the chromosome to a plasmid could result in the spread of intrinsic resistance not only to other strains of E. coli but also to other enterobacterial species.     

Hybrid penicillin-binding proteins in penicillin-resistant strains of Neisseria gonorrhoeae

Benzylpenicillin has been used extensively for approximately 40 years in the treatment of gonorrhoea. The intense selective pressures resulting from the continual exposure of Neisseria gonorrhoeae to penicillin have resulted in the emergence of resistant strains that produce altered forms of penicillin-binding proteins (PBPs) with decreased affinity for the antibiotic. A comparison of the sequences of the PBP-2 genes from penicillin-sensitive and penicillin-resistant strains, suggests that penicillin-resistant forms of PBP 2 may have arisen both by amino-acid substitutions and insertions, and by the exchange of a region encoding part of the penicillin-sensitive transpeptidase domain with the homologous region from a closely related species.     

Whole-genome analysis of diverse Chlamydia trachomatis strains identifies phylogenetic relationships masked by current clinical typing

Chlamydia trachomatis is responsible for both trachoma and sexually transmitted infections, causing substantial morbidity and economic cost globally. Despite this, our knowledge of its population and evolutionary genetics is limited. Here we present a detailed phylogeny based on whole-genome sequencing of representative strains of C. trachomatis from both trachoma and lymphogranuloma venereum (LGV) biovars from temporally and geographically diverse sources. Our analysis shows that predicting phylogenetic structure using ompA, which is traditionally used to classify Chlamydia, is misleading because extensive recombination in this region masks any true relationships present. We show that in many instances, ompA is a chimera that can be exchanged in part or as a whole both within and between biovars. We also provide evidence for exchange of, and recombination within, the cryptic plasmid, which is another key diagnostic target. We used our phylogenetic framework to show how genetic exchange has manifested itself in ocular, urogenital and LGV C. trachomatis strains, including the epidemic LGV serotype L2b.     

Effect of light and pH on diazotized bilirubin

Zusammenfassung Die Wirkung normaler Raumbelichtung (elektrisches fluoreszierendes Licht) und diejenige der Wasserstoffionen-Konzentration auf diazotiertes Bilirubin wurde untersucht. Während elektrisches Licht bei den gegebenen Versuchsbedingungen zu keiner Änderung der optischen Dichte führte, bewirkte ein Zusatz von Natronlauge eine Verminderung der optischen Dichte und zwar unbeeinflusst durch den Zusatz von HCL.     

Localized sex in bacteria

Electrophoretic studies suggest that bacterial populations consist of a number of independent clones and that genetic recombination is rare in nature. But DNA sequencing reveals that individual bacterial genes have a mosaic structure that could have arisen only by recombination. How can these observations be reconciled and what is their relevance to prokaryotic evolution?     

Crustal rheology of the Himalaya and Southern Tibet inferred from magnetotelluric data

The Cenozoic collision between the Indian and Asian continents formed the Tibetan plateau, beginning about 70 million years ago. Since this time, at least 1,400 km of convergence has been accommodated by a combination of underthrusting of Indian and Asian lithosphere, crustal shortening, horizontal extrusion and lithospheric delamination. Rocks exposed in the Himalaya show evidence of crustal melting and are thought to have been exhumed by rapid erosion and climatically forced crustal flow. Magnetotelluric data can be used to image subsurface electrical resistivity, a parameter sensitive to the presence of interconnected fluids in the host rock matrix, even at low volume fractions. Here we present magnetotelluric data from the Tibetan-Himalayan orogen from 77 degrees E to 92 degrees E, which show that low resistivity, interpreted as a partially molten layer, is present along at least 1,000 km of the southern margin of the Tibetan plateau. The inferred low viscosity of this layer is consistent with the development of climatically forced crustal flow in Southern Tibet.