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.     

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.