The herpes simplex virus type 1 genome (160 kilobases) contains three origins of DNA synthesis: two copies of oris located within the repeated sequences flanking the short unique arm (Us), and one copy of OriL located within the long unique arm (UL). Precise localization and characterization of oriL have been severely hampered by the inability to clone sequences which contain it (coordinates 0.398 to 0.413) in an undeleted form in bacteria. We report herein the successful cloning of sequences between 0.398 to 0.413 in an undeleted form, using a yeast cloning vector. Sequence analysis of a 425-base pair fragment spanning the deletion-prone region has revealed a perfect 144-base pair palindrome with striking homology to oris. In a functional assay, the undeleted clone was amplified when functions from herpes simplex virus type 1 were supplied in trans, whereas clones with deletions of 55 base pairs or more were not amplified.The herpes simplex virus type 1 (HSV-1) genome is a 160-kilobase pair (kb) linear duplex DNA molecule consisting of two components, L and S. The L component consists of unique sequences (UL) flanked by the inverted repeated sequences ab and b'a', whereas the S component consists of unique sequences (Us) flanked by inverted repeated sequences ac and c'a' (Fig. 1A) (26,46). The "a" sequence is present as a direct repeat at both molecular termini and in inverted orientation at the L-S junction (9,22,38,56).Attempts to localize the origin(s) of viral DNA synthesis within the structurally complex HSV-1 genome have involved studies of both standard and defective genomes. Electron microscopic studies of replicating standard wildtype viral DNA extracted from infected cells have been interpreted to suggest that the genome contains two origins of viral DNA synthesis, one near the middle of UL and the other near one molecular terminus (20. 27).Indirect evidence supporting the existence of two origins comes from studies of defective molecules of HSV-1 which are generated during serial passage of the virus at high multiplicities of infection. Defective DNA molecules fall into two classes, class I and class II, each consisting of tandem duplications of small subsets of viral DNA sequences. Class I defective genomes contain sequences from the "c" repeats which bracket Us (16-18, 32, 33, 36), whereas class II defective genomes contain sequences from Ul (16,19,32,45) (Fig. IB). Both classes also contain the "a" sequence which specifies a site for the cleavage of viral DNA concatamers during encapsidation (40,55). By analogy with the defective genomes of other DNA viruses, all of which contain origins of DNA synthesis. the existence of two distinct subsets of viral DNA sequences in defective HSV-1 genomes suggests that the genome contains three origins of DNA synthesis: two in diploid "c" sequences (ois), and one in UL (oriL) (Fig. IB).Direct evidence that the repeat units of class I and II * Corresponding author.

SummaryTo date, no prospective study on Plerixafor 'on-demand' in combination with chemotherapy and granulocyte colony-stimulating factor (G-CSF) has been reported. We present an interim analysis of the first prospective study in which Plerixafor was administered on-demand in patients affected by multiple myeloma and lymphoma who received high dose cyclophosphamide or DHAP (dexamethasone, cytarabine, cisplatin) plus G-CSF to mobilize peripheral blood stem cells (PBSC). One hundred and two patients were evaluable for response. A cohort of 240 patients receiving the same mobilizing chemotherapy was retrospectively studied. Failure to mobilize CD34 + cells in peripheral blood was reduced by 'on-demand' strategy compared to conventional mobilization; from 13Á0 to 3Á0% (P = 0Á004). Failure to harvest CD34 + cells 2 9 10 6 /kg decreased from 20Á9 to 4Á0% (P = 0Á0001). The on-demand Plerixafor strategy also resulted in a lower rate of mobilization failure (P = 0Á03) and harvest failure (P = 0Á0008) when compared to a 'bias-adjusted set of controls'. Evaluation of economic costs of the two strategies showed that the overall cost of the two treatments were comparable when salvage mobilizations were taken into account. When in combination with cyclophosphamide or DHAP plus G-CSF, the 'on-demand' use of Plerixafor showed, in comparison to conventionally treated patients, a significant improvement in mobilization of PBSC with no increase in overall cost.

Objective To evaluate the impact of IVUS guidance on the final volume of contrast agent utilized in patients undergoing PCI. Background To date, few approaches have been described to reduce the final dose of contrast agent in percutaneous coronary interventions (PCI). We hypothesized that intravascular ultrasound (IVUS) might serve as an alternative imaging tool to angiography in many steps during PCI, thereby reducing the use of iodine contrast. Methods A total of 83 patients were randomized to I) angiography-guided PCI or II) IVUS-guided PCI, both groups treated according to a pre-defined meticulous procedural strategy. The primary endpoint was the total volume contrast agent used during PCI. Patients were followed clinically for an average of 4 months. Results The median total volume of contrast was 64.5 ml (interquartile range [IQR] 42.8 – 97.0 ml; minimum 19 ml; maximum 170 ml) in angiography-guided group vs. 20.0 ml (IQR 12.5 – 30.0 ml; minimum 3 ml; maximum 54 ml) in IVUS-guided group (p<0.001). Similarly, the median volume of contrast / creatinine clearance ratio was significantly lower among patients treated with IVUS-guided PCI (1.0 [IQR 0.6 – 1.9] vs. 0.4 [IQR 0.2 – 0.6] respectively; p<0.001). In-hospital and 4-month outcomes were not different between patients randomized to angiography-guided and IVUS-guided PCI. Conclusions Thoughtful and extensive utilization of IVUS as the primary imaging tool to guide PCI is safe, and markedly reduces the volume of iodine contrast, compared to angiography-alone guidance. The use of IVUS should be considered for patients at high risk for contrast-induced acute kidney injury or volume overload undergoing coronary angioplasty.

Artificial addition of calcium-like elements to the atherosclerotic plaque led to an increase in necrotic tissue in virtual histology that is probably artefactual. The overestimation of necrotic tissue by calcium strictly followed a linear pattern, indicating that it may be amenable to mathematical correction.

No abstract