Pore formation by complement in the outer membrane of Gram-negative bacteria studied with asymmetric planar lipopolysaccharide/phospholipid bilayers

Summary The interaction of complement with an asymmetric planar lipopolysaccharide/phospholipid bilayer system as a model for the lipid matrix of the outer membrane of Gram-negative bacteria has been studied. The addition of whole human serum to the aqueous solution at the lipopolysaccharide side of the asymmetric membrane resulted in a rapid increase of the bilayer conductance in discrete steps, indicating the formation of transmembrane pores, which were not observed in the case of pure phospholipid membranes. The amplitudes of the discrete conductance steps varied over a range of more than one order of magnitude. The mean single step conductance was (0.39±0.24) nS for a subphase containing (inmm): 100 KCl, 5 MgCl₂ and 5 HEPES buffer. The steps were grouped into bursts of typically 9±3 events per burst and the conductance change within one burst was (8.25±4.00) nS.The pore-forming activity of serum at the asymmetric membrane system was independent of the presence of specific antibodies against the lipopolysaccharide but was dependent on calcium ions. Furthermore, the pore-forming activity required complement component C9.A model for the mode of pore formation by complement is proposed: The complement pore is generated in discrete steps by insertion of C9 monomers into the membrane and their irreversible aggregation to water-filled channels with a diameter of approximately 7 nm assuming a circular geometry.     

Newly discovered populations of the Ethiopian endemic and endangered Afrixalus clarkei Largen,implications for conservation

Knowledge of the Ethiopian amphibian fauna is limited and Southwest Ethiopia remains understudied. This part of Ethiopia, where most of the country’s remaining natural forest is situated, is known to harbour the only populations of Afrixalus clarkei (Largen), an endemic banana frog, worldwide. This species is under great threat of extinction and is therefore classified as endangered on the IUCN red list. We surveyed different potential habitats for this species outside its known range and found several new populations extending its known habitat preference, and the geographical and altitudinal range of the species. We here show that Afrixalus clarkei is more common than previously thought.     

Impaired High-Density Lipoprotein Anti-Oxidant Function Predicts Poor Outcome in Critically Ill Patients

Introduction

Oxidative stress affects clinical outcome in critically ill patients. Although high-density lipoprotein (HDL) particles generally possess anti-oxidant capacities, deleterious properties of HDL have been described in acutely ill patients. The impact of anti-oxidant HDL capacities on clinical outcome in critically ill patients is unknown. We therefore analyzed the predictive value of anti-oxidant HDL function on mortality in an unselected cohort of critically ill patients.

Method

We prospectively enrolled 270 consecutive patients admitted to a university-affiliated intensive care unit (ICU) and determined anti-oxidant HDL function using the HDL oxidant index (HOI). Based on their HOI, the study population was stratified into patients with impaired anti-oxidant HDL function and the residual study population.

Results

During a median follow-up time of 9.8 years (IQR: 9.2 to 10.0), 69% of patients died. Cox regression analysis revealed a significant and independent association between impaired anti-oxidant HDL function and short-term mortality with an adjusted HR of 1.65 (95% CI 1.22–2.24; p = 0.001) as well as 10-year mortality with an adj. HR of 1.19 (95% CI 1.02–1.40; p = 0.032) when compared to the residual study population. Anti-oxidant HDL function correlated with the amount of oxidative stress as determined by Cu/Zn superoxide dismutase (r = 0.38; p<0.001).

Conclusion

Impaired anti-oxidant HDL function represents a strong and independent predictor of 30-day mortality as well as long-term mortality in critically ill patients.     

Quantitative Cytochemische Untersuchungen an Nukleolen aus Speicheldrüsenkernen von Chironomus Thummi

Zusammenfassung Trockengewicht und Absorptionsverhältnisse des Nukleolus am 4. Speicheldrüsenchromosom von Chironomus thummi wurden mikrointerferometrisch und mikrospektrophotometrisch bestimmt.Die Trockengewichtszunahme des Nukleolus während der Larvenentwicklung folgt weder linear noch exponentiell der zunehmenden Polytänie der Chromosomen. Gegen Ende des 3. Larvenstadiums steigen die Trockengewichte der Nukleolen unabhängig vom Polytäniegrad individueller Chromosomen steil an.Das Verhältnis von Ribonukleinsäuren zu Proteinen bleibt in den Nukleolen von Speicheldrüsenkernen aller Larvenstadien bis zur Degenerierung der Nukleoli trotz bedeutender Veränderungen ihrer Totalmasse konstant.Induzierte Störungen des Energiestoffwechsels der Larven durch Hungern und Übervölkerung in den Kulturschalen führen zu erheblicher Rückbildung der Nukleolarmasse — bzw. zu Hemmung in der Akkumulation von Nukleolarsubstanz -, ohne die Konstanz der Ribonukleoproteidverhältnisse zu ändern. Ebenso bewirkt Kälteschock zwar eine Kontraktion der Nukleolen, doch keine Änderung in der Zusammensetzung der Grundkomponenten.Mit Unterstützung des Schwedischen Medizinischen Forschungsrates, der Wallenberg-Stiftung und des U.S. Public Health Service (Grant C-3082).