Bacterial glycosidases for the production of universal red blood cells.

Enzymatic removal of blood group ABO antigens to develop universal red blood cells (RBCs) was a pioneering vision originally proposed more than 25 years ago. Although the feasibility of this approach was demonstrated in clinical trials for group B RBCs, a major obstacle in translating this technology to clinical practice has been the lack of efficient glycosidase enzymes. Here we report two bacterial glycosidase gene families that provide enzymes capable of efficient removal of A and B antigens at neutral pH with low consumption of recombinant enzymes. The crystal structure of a member of the alpha-N-acetylgalactosaminidase family reveals an unusual catalytic mechanism involving NAD+. The enzymatic conversion processes we describe hold promise for achieving the goal of producing universal RBCs, which would improve the blood supply while enhancing the safety of clinical transfusions.

Pathway prediction by bioinformatic analysis of the untranslated regions of the CFTR mRNA.

Mining the information contained within the genetic code in untranslated regions has proven difficult because of the ambiguity of microRNA and protein binding sites. This manuscript describes a bioinformatic screen that identifies long sequences with partial identity to the untranslated regions of the cystic fibrosis transmembrane regulator. This screen uncovered a long, evolutionarily conserved motif common to the 3' UTRs of the CFTR and SEC24A transcripts, and shorter, statistically significant motifs unique to either 5' or 3' UTRs. In addition, of the 140 transcripts identified in the screen that encode proteins with known protein interactions, 130 are linked to CFTR through protein interactions. The screen identified genes that are known to be involved in lung fibrosis, the inflammatory response of cystic fibrosis and sensitivity to Pseudomonas aeruginosa infections. The bioinformatic analysis of untranslated regions should prove to be a powerful adjunct to other tools for predicting pathways and relevant interactions.

Miner Artist/Minor Artist? Class, Politics, and the Post-industrial Consumption of Mining Art.

This article uses the recently discovered art work of a County Durham coal miner, Jimmy Kays (1886-1951) to highlight the terms in which coal mining art has achieved popularity and value in the post-mining period. Kays' work is considered with reference to the presenting narrative that promotes and markets mining art not only in terms of its intrinsic artistic quality, but also as a desirable commodity which, as a legacy of the mining past, can contribute to the revival of post-mining places. Maximizing the value that can accrue from mining art in post-industrial conditions involves appealing to the interest of the largest possible audience. The consequence of this is the dominance of a particular interpretation of the mining past. The art of Jimmy Kays does not conform to the conditions of the market, and cannot achieve a similar status. Despite its artistic qualities and its uniqueness as the product of a Durham working miner in the early twentieth century, it sits outside the dominant lexicon of coal mining art. The outsider status of Jimmy Kays is an example of a wider set of issues relating to the invisibility of working class creativity and the difficulties of achieving excellence or public acknowledgment in conditions that lack organizational support and in which value is established elsewhere. I argue that an understanding of the invisibility of art work such as that produced by Kays illuminates the exercise of class-based power in terms of the production, consumption, and range of meaning inscribed within popular mining art. Mining art that has been allocated value is in danger of being appropriated in ways that pacify rather than energize audiences, by foregrounding particular aspects of the mining past for purposes of consumption whilst submerging the issues that link more troubled aspects of the past with the present.

Messenger RNAs under differential translational control in Ki-ras-transformed cells.

Microarrays have been used extensively to identify differential gene expression at the level of transcriptional control in oncogenesis. However, increasing evidence indicates that changes in translational control are critical to oncogenic transformation. This study identifies mRNA transcripts that are differentially regulated, primarily at the level of translation, in the immortalized human embryonic prostate epithelial cell line 267B1 and the v-Ki-ras-transformed counterpart by comparing total mRNA to polysome-bound mRNA by using Affymetrix oligonucleotide microarrays. Among the transcripts that were identified were those encoding proteins involved in DNA replication, cell cycle control, cell-to-cell interactions, electron transport, G protein signaling, and translation. Many of these proteins are known to contribute to oncogenesis or have the potential to contribute to oncogenesis. Differential expression of RNA-binding proteins and the presence of highly conserved motifs in the 5' and 3' untranslated regions of the mRNAs are consistent with multiple pathways and mechanisms governing the changes in translational control. Although Alu sequences were found to be associated with increased translation in transformed cells, an evolutionarily conserved motif was identified in the 3' untranslated regions of ephrinB1, calreticulin, integrin alpha3, and mucin3B that was associated with decreased polysome association in 267B1/Ki-ras.

Computational prediction of the polyQ and CAG repeat spinocerebellar ataxia network based on sequence identity to untranslated regions.

Computational prediction of biological networks would be a tremendous asset to systems biology and personalized medicine. In this paper, we use a moving window bioinformatic screen to identify transcripts with partial identity to the 5' and 3'UTRs of the polyQ spinocerebellar ataxia (SCA) genes ATXN1, ATXN2, ATXN3, ATXN7, TBP and CACNA1A and the CAG repeat expansion gene PPP2R2B. We find that the bioinformatic screen enriches for transcripts that encode proteins that interact and that have functions relevant to polyQ SCA. Transcription control and RNA binding are the primary functional groups represented in the proteins from the combined screens. The insulin growth factor pathway, the WNT pathway, long term potentiation, melanogenesis and ATM mediated DNA repair pathways were identified as important pathways. UGUUU repeats were identified as an abundant motif in the SCA network and PAXIP1, CELF2, CREBBP, EBF1, PLEKHG4, SRSF4, C5orf42, NFIA, STK24, and YWHAG were identified as statistically significant proteins in the polyQ and PPP2R2B network.

Identification of a GH110 subfamily of alpha 1,3-galactosidases: novel enzymes for removal of the alpha 3Gal xenotransplantation antigen.

In search of alpha-galactosidases with improved kinetic properties for removal of the immunodominant alpha1,3-linked galactose residues of blood group B antigens, we recently identified a novel prokaryotic family of alpha-galactosidases (CAZy GH110) with highly restricted substrate specificity and neutral pH optimum (Liu, Q. P., Sulzenbacher, G., Yuan, H., Bennett, E. P., Pietz, G., Saunders, K., Spence, J., Nudelman, E., Levery, S. B., White, T., Neveu, J. M., Lane, W. S., Bourne, Y., Olsson, M. L., Henrissat, B., and Clausen, H. (2007) Nat. Biotechnol. 25, 454-464). One member of this family from Bacteroides fragilis had exquisite substrate specificity for the branched blood group B structure Galalpha1-3(Fucalpha1-2)Gal, whereas linear oligosaccharides terminated by alpha1,3-linked galactose such as the immunodominant xenotransplantation epitope Galalpha1-3Galbeta1-4GlcNAc did not serve as substrates. Here we demonstrate the existence of two distinct subfamilies of GH110 in B. fragilis and thetaiotaomicron strains. Members of one subfamily have exclusive specificity for the branched blood group B structures, whereas members of a newly identified subfamily represent linkage specific alpha1,3-galactosidases that act equally well on both branched blood group B and linear alpha1,3Gal structures. We determined by one-dimensional (1)H NMR spectroscopy that GH110 enzymes function with an inverting mechanism, which is in striking contrast to all other known alpha-galactosidases that use a retaining mechanism. The novel GH110 subfamily offers enzymes with highly improved performance in enzymatic removal of the immunodominant alpha3Gal xenotransplantation epitope.

Secondary prevention of vitamin D-deficiency rickets.

Reports of vitamin D-deficiency rickets and its associated morbidity continue among inadequately supplemented, dark-skinned breastfed infants. Despite the new vitamin D dietary guidelines, there remain significant numbers of unsupplemented breastfed infants. Here we report a case of subclinical vitamin D-deficiency rickets. This patient had biochemical and radiographic but not clinical evidence for rickets. We propose a new step of screening high-risk infants for subclinical rickets using wrist films paired with 25-hydroxyvitamin D levels.