Neutrophil priming mechanisms of sulfolipid-I and n-formyl-methionyl-leucyl-phenylalanine

The principal sulfatide of virulentMycobacterium tuberculosis, sulfolipid-I (SL-I), both directly stimulates neutrophil superoxide (O ₂ ^– ) release and, at substimulatory concentrations, primes these cells for markedly enhanced oxidative responsiveness to other stimuli. The present study was undertaken to clarify the priming mechanisms by comparing cellular events following priming doses of SL-I with those following priming with N-formyl-methionyl-leucyl-phenylalanine (FMLP). We compared the involvement of the calcium cation (Ca²⁺), as well as membrane protein kinase C (PKC) activity and the translocation of NADPH oxidase-cytosolic cofactor effected by priming levels of the two agonists. The investigation led to two important conclusions. First, we clearly demonstrate that priming by both SL-I and FMLP results from activation of cellular processes that are not involved in direct oxidative activation. For example, whereas direct induction of O ₂ ^– generation by FMLP and SL-I required increases in intracellular Ca²⁺, an increase in intracellular calcium concentration ([Ca²⁺]_i) above basal levels was not required for priming. Second, we identified key differences in the cellular responses to priming doses of SL-I and FMLP. Whereas increased membrane PKC activity caused by priming doses of FMLP was only partially blocked by chelation of intracellular Ca²⁺, Ca²⁺ chelation completely inhibited the increase in membrane PKC activity caused by SL-I. NADPH oxidase-cytosolic factor translocation to plasma membranes was completely blocked by pertussis toxin when priming doses of SL-I were used. This guanine-nucleotide-binding protein inhibitor had no effect on FMLP-dependent translocation of the oxidase cofactors. The comparative approach introduced in this report provides a valuable and novel method to discern the complex interactions of various cellular processes that regulate the state of activation of stimulated cells.     

Diversity in nucleotide acquisition by antigenically similar Chlamydia psittaci of avian origin

Abstract Two different nucleic acid precursor utilization patterns were obtained for five avian isolates of Chlamydia psittaci . Three of the isolates bahaved in a manner similar to that previously described, showing total dependency on the host cell for ribonucleoside triphosphates and being unable to utilize medium-supplied thymidine. In contrast, the other two isolates were incapable of taking pyrimidine ribonucleotides from the host cell and they could efficiently utilize medium-supplied thymidine. These unusual isolates were resistant to 5-fluorouridine while the other three isolates were sensitive. Of the five isolates only 6BC was sensitive to sulfonamides. The five isolates were divided into two groups by comparing the Alu I restriction endonuclease patterns obtained following digestion of the major outer membrane protein (OMP1) gene, amplified by the polymerase chain reaction. The OMP1 genotyping results were confirmed by serotyping.     

A NEW PHYLOGENY FOR CHROMOPHYTE ALGAE USING 16S-LIKE RRNA SEQUENCES FROM MALLOMONAS PAPILLOSA (SYNUROPHYCEAE) AND TRIBONEMA AEQUALE (XANTHOPHYCEAE)1

The 16S-like ribosomal RNA genes from Mallomonas papillosa Harris et Bradley (Synurophyceae) and Tribonema aequale Pascher (Xanthophyceae) were sequenced and compared to those of other eukaryotes. Mallomonas is closely related to Ochromonas (Chrysophyceae) and supports the general hypothesis of a close phylogenetic relationship between the Synurophyceae and Chrysophyceae. Tribonema is specifically related to Costaria costata (C. A. Agardh) Saunders (Phaeophyceae) demonstrating an unexpected phylogenetic relationship between the Xanthophyceae and Phaeophyceae. Distance and parsimony analysis place these four chromophyte genera in a complex evolutionary assemblage that includes the Bacillariophyceae and Oomycetes but excludes the Dinophyceae. The close relationship between the chromophyte algae and the Öomycete fungi supports the hypothesis that protists with tripartite hairs form a natural assemblage.     

COMPETITION BETWEEN STAURASTRUM LUETKEMUELLERII (CHLOROPHYCEAE) AND MICROCYSTIS AERUGINOSA(CYANOPHYCEAE) UNDER VARYING MODES OF PHOSPHATE SUPPLY1

The desmid Staurastrum luetkemuellerii Donat et Ruttner and the cyanobacterium Microcystis aeruginosa Kütz. were grown in mixed cultures with various phosphate (P_i) additions. One pulse of P_i each day (semi-continuous cultures) favored M. aeruginosa whereas S. luetkemuellerii was favored when the same quantity of P_i was supplied continuously (chemostats). Both species coexisted under P limitation provided that the nutrient was supplied in an appropriate mode. The ability of each species to compete for P depended on their P_i uptake characteristics and their capability to retain the accumulated P_i. High affinity in uptake at low P_i concentrations contributed considerably to the growth eficiency of S. luetkemuellerii under continuous supply of P_iM. aeruginosa was, however, consistently superior to S. luetkemuellerii in accuniulatiug the newly added P, but had a high rate of P_i release. In both -types of cultures, a net high of P went from M. aeruginosa to S. luetkemuellerii. The kinetic characteristics of the two species were used to simulate the outcome of competition experiments. Simulations agreed with the experimental data f both uptake and P_i release were considered in the model. The zlariable P*(the concentration of P_i at which the net uptake is equal to μ·Q_P is a function of uptake and release of P_i but could not explain the chemostat results. S. luetkemuellerii was the winner in many experiments even if its P*was higher thou that of M. aeruginosa. Thus, in the present case P_c (the concentration at which the net uptake is zero) was a better predictor of the ability to compete for P_i under steady state as well as transient conditions in the P_i supply.     

A Sperm Midpiece Defect in a Hereford Bull with Variable Semen Quality and Freezability

A midpiece sperm defect with a frequency of 25-35% in ejaculates obtained from a Hereford bull with a 60 d non-retum rate of 76.4% after careful pre- and postfreeze semen selection was studied in light microscope and by transmission electron microscopy. The defect consisted in a folding and coiling of the distal midpiece characterized by disorganization and irregularity of mitochondria surrounding the axial fiber bundle, combined with retraction of doublet fibers and dislocation and fracturing of these elements and the corresponding dense fibers. Based on examination of the sper- matogenic epithelium it was concluded that the alterations in the axial fiber bundle were secondary to those in the mitochondrial sheath. The abnormality appeared to be related to the “Dag-like” defect earlier observed in different breeds.     

Double blind randomised controlled trial of effect of metoprolol on myocardial ischaemia during endoscopic cholangiopancreatography.

OBJECTIVE--To evaluate the effect of metoprolol, a beta adrenergic blocking drug, on the occurrence of myocardial ischaemia during endoscopic cholangiopancreatography. DESIGN--Double blind, randomised, controlled trial. SETTING--University Hospital. SUBJECTS--38 (two groups of 19) patients scheduled for endoscopic cholangiopancreatography. INTERVENTIONS--Metoprolol 100 mg or placebo as premedication two hours before endoscopy. MAIN OUTCOME MEASURES--Heart rate, arterial oxygen saturation by continuous pulse oximetry, ST segment changes during endoscopic cholangiopancreatography (an ST segment deviation > 1 mV was defined as myocardial ischaemia), electrocardiogram monitored continuously with a Holter tape recorder. RESULTS--All patients had increased heart rate during endoscopy compared with rate before endoscopy, but heart rate during endoscopy was significantly lower in the metoprolol group compared with the placebo group (P = 0.0002). Twenty one patients (16 placebo, 5 metoprolol; P = 0.0008) developed tachycardia (heart rate > 100/min) during the procedure, and 11 patients (10 placebo, 1 metoprolol; P = 0.003) developed myocardial ischaemia. One patient in the placebo group had an acute inferolateral myocardial infarction. In the 10 other patients with signs of myocardial ischaemia during endoscopy the ST deviation disappeared when the endoscope was retracted. In all patients myocardial ischaemia was related to increases in heart rate, and 10 of the 11 patients had tachycardia coherent with myocardial ischaemia. CONCLUSIONS--Metoprolol prevented myocardial ischaemia during endoscopic cholangiopancreatography, probably through lowering the heart rate. Thus, tachycardia seems to be a key pathogenic factor in the development of myocardial ischaemia during endoscopy.     

Identification of a locus involved in meningococcal lipopolysaccharide biosynthesis by deletion mutagenesis

A novel method for insertion/deletion mutagenesis in meningococci was devised. This consisted of ligating a digest of total chromosomal DNA to a 1.1 kb restriction fragment containing an erythromycin-resistance marker ( ermC ), and subsequent transformation of the ligation mixture into the homologous meningococcal strain H44/76. Southern blotting of a number of the resulting erythromycin-resistant transformants demonstrated that all carried the ermC gene inserted at different positions in the chromosome. Mutants with a specific phenotype were identified by screening with the anti-lipopolysaccharide (LPS) monoclonal antibody MN4A8B2, which is specific for immunotype L3. In this way, two independent L3-negative mutant strains were isolated. In transformation experiments with chromosomal DNA from these mutants, erythromycin-resistance and lack of MN4A8B2 reactivity were always linked, showing that the insertion/deletion was in a locus involved in LPS biosynthesis. On SDS–PAGE, the mutant LPS displayed an electrophoretic mobility intermediate between that produced by the previously isolated galE and rfaF mutant strains. Chemical analysis of the mutant LPS revealed that the structure was probably lipid A–(KDO)₂–(Hep)₂. Chromosomal DNA flanking the ermC insertion in these two mutant strains was cloned, and used as probe for the isolation of the corresponding region of the wild-type strain. From hybridization and polymerase chain reaction (PCR) analysis, it could be concluded that both mutations map to the same locus. The affected gene probably encodes the glycosyltransferase necessary for adding N -acetylglucosamine to heptose.     

Succession and soil development in man-made coastal ecosystems at the Baltic Sea

By the creation of Køge Bay Seaside Park in 1978 the opportunity was given to study the establishment of vegetation and primary succession in a man-made coastal area. As the soil consists of marine material from the Baltic Sea, no organic matter or seedbank was present. The first steps of primary succession were followed in 1980, when the initial inventories of flora immigration and soil development were carried out in coastal grasslands and plantings. The surveys were repeated in 1992 and 1993. Except for the planted woody species and a few sown grasses, all other species of plants have reached the area through natural dispersal of diaspores. The total number of species in the permanent plots has increased from 26 in 1980 to 91 in 1993. The results indicate that this number will continue to increase in the coming years until a certain level. Then it will probably decrease as a result of competition from woody species, unless the vegetation is kept in a steady state by disturbances or management. Today the area is very far from the initial situation, and the off-shore barrier has changed towards a landscape dominated by small groves and grasslands of an urban common type.