Structural determinants and distribution of phosphate specificity in ribonucleotide reductases.

Ribonucleotide reductases (RNRs) catalyze the reduction of ribonucleotides to the corresponding deoxyribonucleotides, the building blocks of DNA. RNRs are specific for either ribonucleoside diphosphates or triphosphates as substrates. As far as is known, oxygen-dependent class I RNRs (NrdAB) all reduce ribonucleoside diphosphates, and oxygen-sensitive class III RNRs (NrdD) are all ribonucleoside triphosphate reducers, whereas the adenosylcobalamin-dependent class II (NrdJ) contains both ribonucleoside diphosphate and triphosphate reducers. However, it is unknown how this specificity is conveyed by the active site of the enzymes and how this feature developed in RNR evolution. By structural comparison of the active sites in different RNRs, we identified the apical loop of the phosphate-binding site as a potential structural determinant of substrate specificity. Grafting two residues from this loop from a diphosphate- to a triphosphate-specific RNR caused a change in preference from ribonucleoside triphosphate to diphosphate substrates in a class II model enzyme, confirming them as the structural determinants of phosphate specificity. The investigation of the phylogenetic distribution of this motif in class II RNRs yielded a likely monophyletic clade with the diphosphate-defining motif. This indicates a single evolutionary-split event early in NrdJ evolution in which diphosphate specificity developed from the earlier triphosphate specificity. For those interesting cases where organisms contain more than one nrdJ gene, we observed a preference for encoding enzymes with diverse phosphate specificities, suggesting that this varying phosphate specificity confers a selective advantage.

Clinical experience of photon counting breast tomosynthesis: comparison with traditional mammography.

BACKGROUND: In two-dimensional mammography, a well-known problem is over- and underlying tissue which can either obstruct a lesion or create a false-positive result. Tomosynthesis, with an ability to layer the tissue in the image, has the potential to resolve these issues. PURPOSE: To compare the diagnostic quality, sensitivity and specificity of a single tomosynthesis mammography image and a traditional two-view set of two-dimensional mammograms and to assess the comfort of the two techniques. MATERIAL AND METHODS: One hundred and forty-four women, mainly chosen because of suspicious features on standard mammograms (76 malignant), had a single tomosynthesis image taken of one breast using a novel photon counting system. On average, the dose of the tomosynthesis images was 0.63 times that of the two-view images and the compression force during the procedure was halved. The resulting images were viewed by two radiologists and assessed both individually and comparing the two techniques. RESULTS: In 56% of the cases the radiologists rated the diagnostic quality of the lesion details higher in the tomosynthesis images than in the conventional images (and in 91% equal or higher), which means there is a statistically significant preference for the tomosynthesis technique. This included the calcifications which were rated as having better quality in 41% of the cases. While sensitivity was slightly higher for traditional mammography the specificity was higher for tomosynthesis. However, neither of these two differences was large enough to be statistically significant. CONCLUSION: The overall accuracy of the two techniques was virtually equal despite the radiologist's very limited experience with tomosynthesis images and vast experience with two-dimensional mammography. As the diagnostic quality of the lesion details in the tomosynthesis images was valued considerably higher this factor should improve with experience. The patients also favored the tomosynthesis examination, rating the comfort of the procedure as much higher than regular mammography which might affect screening attendance.