Resolution and characterisation of multiple isoforms of bovine kappa-casein by 2-DE following a reversible cysteine-tagging enrichment strategy

Visualisation of multiple isoforms of kappa-casein on 2-D gels is restricted by the abundant alpha- and beta-caseins that not only limit gel loading but also migrate to similar regions as the more acidic kappa-casein isoforms. To overcome this problem, we took advantage of the absence of cysteine residues in alpha(S1)- and beta-casein by devising an affinity enrichment procedure based on reversible biotinylation of cysteine residues. Affinity capture of cysteine-containing proteins on avidin allowed the removal of the vast majority of alpha(S1)- and beta-casein, and on subsequent 2-D gel analysis 16 gel spots were identified as kappa-casein by PMF. Further analysis of the C-terminal tryptic peptide along with structural predictions based on mobility on the 2-D gel allowed us to assign identities to each spot in terms of genetic variant (A or B), phosphorylation status (1, 2 or 3) and glycosylation status (from 0 to 6). Eight isoforms of the A and B variants with the same PTMs were observed. When the casein fraction of milk from a single cow, homozygous for the B variant of kappa-casein, was used as the starting material, 17 isoforms from 13 gel spots were characterised. Analysis of isoforms of low abundance proved challenging due to the low amount of material that could be extracted from the gels as well as the lability of the PTMs during MS analysis. However, we were able to identify a previously unrecognised site, T-166, that could be phosphorylated or glycosylated. Despite many decades of analysis of milk proteins, the reasons for this high level of heterogeneity are still not clear.

An analysis of the images attached to referral messages in an e-mail based telemedicine system for developing countries

Little is known about the quality of the images transmitted in email telemedicine systems. The present study was designed to survey the quality of images transmitted in the Swinfen Charitable Trust email referral system. Telemedicine cases were examined for a 3 month period in 2002 and a 3 month period in 2006. The number of cases with images attached increased from 8 (38%) to 37 (53%). There were four types of images (clinical photographs, microscope pictures, notes and X-ray images) and the proportion of radiology images increased from 27 to 48%. The cases in 2002 came from four different hospitals and were associated with seven different clinical specialties. In 2006, the cases came from 19 different hospitals and 20 different specialties. The 46 cases (from both study periods) had a total of 159 attached images. The quality of the images was assessed by awarding each image a score in four categories: focus, anatomical perspective, composition and lighting. The images were scored on a five-point scale (1 = very poor to 5 =very good) by a qualified medical photographer. In comparing image quality between the two study periods, there was some evidence that the quality had reduced, although the average size of the attached images had increased. The median score for all images in 2002 was 16 (interquartile range 14-19) and the median score in 2006 was 15 (13-16). The difference was significant (P < 0.001, Mann-Whitney test).