Lanthanide probes in biological systems: characterization of luminescence excitation spectra of terbium complexes with proteins

Upon irradiation in the ultraviolet region aromatic chromophores may transfer energy to a nearby Tb³⁺, which in turn emits a green phosphorescence. This paper reports the characterization of the ultraviolet excitation spectra of aromatic chromophores capable of transferring energy to Tb³⁺ by monitoring of the green Tb³⁺ emission in the 540-550 nm region. Results are included for complexes containing phenyl, hydroxyphenyl, indole. and catechol chromophores. Characteristic excitation spectra are presented for the aromatic chromophores occurring as side chains in proteins. Though it is preferable to compare entire excitation spectra, the ratio of intensities at 292 to 276 nm, R, is suggested as a useful diagnostic criterion. Numerical R values are indicative of the following aromatic side chains as the energy donor to Tb³⁺: R <0.2, unionized tyrosine; R = 0.5 to 1.0, tryptophan; and R > 1.8. ionized tyrosine. Tlie phenylalanyl chromophore displays a definitive excitation spectrum at shorter wavelengths. For ovotransferrin R = 0.9 and comparison of the full excitation spectra suggests that it contains comparable contributions from both ionized tyrosine and tryptophan side chains. Some difficulties in obtaining reliable excitation spectra are described. An analysis of inner-filtering of incident light reveals that for an absorbance less than 0.8 the excitation spectrum is broadened and flattened compared to the absorption spectrum. At maximum absorbances greater than 0.8 false maxima may appear to both sides of a real maximum. Two spurious maxima in an excitation spectrum were generated in a Tb³⁺ complex and compared to the correct excitation spectrum of the same complex obtained at lower absorbance.     

Molecular Epidemiology of Carbapenem Non-Susceptible Acinetobacter baumannii in France

Carbapenem-resistant Acinetobacter baumannii have emerged globally. The objective of this study was to investigate the epidemiology, clonal diversity and resistance mechanisms of imipenem non-susceptible A. baumannii isolates in France. Between December 2010 and August 2011, 132 notifications were collected, including 37 outbreaks corresponding to 242 cases (2 to 55 per cluster). Multilocus sequence typing, pulsed-field gel electrophoresis (PFGE) and characterisation of carbapenemase-encoding genes were performed on 110 non-repetitive isolates. Gene bla _OXA-23 was the most frequently detected (82%), followed by bla _OXA-24 (11%) and bla _OXA-58 (7%). Eleven sequence types (ST) were distinguished, among which sequence types ST1, ST2 (64%), ST20, ST25, ST85 and ST107. Isolates from epidemiological clusters had the same ST and resistance genes, indicating probable transmission within centres. In contrast, PFGE types of isolates differed among centres, arguing against transmission among centers. This study provides the first epidemiological snapshot of the population of A. baumannii with reduced susceptibility to carbapenems from France, and further underlines the predominance of international clones.     

Should doctors be more proactive as advocates for victims of violence?

Currently the management of adult victims of violence by general practitioners and accident and emergency departments is reactive, concerned almost exclusively with the management of physical injuries. Professor Jonathan Shepherd outlines some ideas for a more proactive approach on the part of doctors to improve the protection and support of vulnerable people; to deal with psychological sequelae; to take the responsibility of making an official complaint to the police away from seriously injured people, who are unable to give or withhold consent to disclosure; and to prevent assailants inflicting further injuries. We asked a sociologist, a psychiatrist, a moral philosopher, and a police surgeon for their comments.     

Hydrolysis of diacylglycerols by lipoprotein lipase.

Enantiomeric diacylglycerols were emulsified, mole for mole, with lyso(1-acyl) lecithin and were hydrolyzed with lipoprotein lipase in NH4Cl-beef serum albumin buffer at pH 8.6 after a brief incubation with delipidated rat serum. The enzyme was prepared from lyophilized and dialyzed bovine skim milk in a 4 percent solution. The course of hydrolysis for each set of enantiomers was determined by gas-liquid chromatography of the masses of the diacylglycerols remaining or monoacylglycerols released in the medium between 0 and 15 min. The majority of sets of sn-1,2- and 2,3-diacylglycerols, including an isotope-labeled true enantiomeric set which was assessed by mass spectrometry, demonstrated preference by the enzyme for lipolysis at position 1 but with less specificity than previously was shown in sn-triacylglycerol hydrolysis. The results preclude the possibility that the predominance of sn-2,3-diacylglycerol intermediates during triacylglycerol hydrolysis is due solely to a preferential breakdown of the 1,2-isomers and reinforce the conclusion that lipoprotein lipase is specific for position 1.     

Prior evolution in stochastic versus constant temperatures affects RNA virus evolvability at a thermal extreme

It is unclear how historical adaptation versus maladaptation in a prior environment affects population evolvability in a novel habitat. Prior work showed that vesicular stomatitis virus (VSV) populations evolved at constant 37°C improved in cellular infection at both 29°C and 37°C; in contrast, those evolved under random changing temperatures between 29°C and 37°C failed to improve. Here, we tested whether prior evolution affected the rate of adaptation at the thermal‐niche edge: 40°C. After 40 virus generations in the new environment, we observed that populations historically evolved at random temperatures showed greater adaptability. Deep sequencing revealed that most of the newly evolved mutations were de novo. Also, two novel evolved mutations in the VSV glycoprotein and replicase genes tended to co‐occur in the populations previously evolved at constant 37°C, whereas this parallelism was not seen in populations with prior random temperature evolution. These results suggest that prior adaptation under constant versus random temperatures constrained the mutation landscape that could improve fitness in the novel 40°C environment, perhaps owing to differing epistatic effects of new mutations entering genetic architectures that earlier diverged. We concluded that RNA viruses maladapted to their previous environment could “leapfrog” over counterparts of higher fitness, to achieve faster adaptability in a novel environment.