Differential divalent cation requirements uncouple the assembly and catalytic reactions of human immunodeficiency virus type 1 integrase

Previous in vitro analyses have shown that the human immunodeficiency virus type 1 (HIV-1) integrase uses either manganese or magnesium to assemble as a stable complex on the donor substrate and to catalyze strand transfer. We now demonstrate that subsequent to assembly, catalysis of both 3' end processing and strand transfer requires a divalent cation cofactor and that the divalent cation requirements for assembly and catalysis can be functionally distinguished based on the ability to utilize calcium and cobalt, respectively. The different divalent cation requirements manifest by these processes are exploited to uncouple assembly and catalysis, thus staging the reaction. Staged 3' end processing and strand transfer assays are then used in conjunction with exonuclease III protection analysis to investigate the effects of integrase inhibitors on each step in the reaction. Analysis of a series of related inhibitors demonstrates that these types of compounds affect assembly and not either catalytic process, therefore reconciling the apparent disparate results obtained for such inhibitors in assays using isolated preintegration complexes. These studies provide evidence for a distinct role of the divalent cation cofactor in assembly and catalysis and have implications for both the identification and characterization of integrase inhibitors.     

In times of survival, the right of silence

The purpose of this study was to assess cortical and cancellous bone responses to unilateral limb immobilization and, subsequently, to remobilization with exercise, in a young adult canine model. Right forelimbs of 14 1-2-year old mongrel dogs were immobilized in a non-weight-bearing position by a bandage for 16 weeks. Six control dogs were untreated. At 16 weeks, seven immobilized and three control dogs were euthanized. The remaining seven immobilized dogs began a recovery protocol consisting of 16 weeks of kennel confinement (without the right forelimb bandaged) followed by 16 weeks of treadmill exercise conducted three times per week. These seven dogs and three control dogs were euthanized at 48 weeks. Bone mineral density of the proximal radii was determined with dual-energy X-ray absorptiometry and humeral middiaphyseal cross-sectional areas were determined with computed tomography. Humeri were tested in cranio-caudal three-point bending to failure. Cancellous bone cores from the lateral humeral condyles had wet apparent density determined and were tested to failure in compression. Mechanical properties, bone density, and cross-sectional areas were compared between immobilized (right forelimb), contralateral weight bearing (left forelimb), and control forelimbs with Kruskal-Wallis and post hoc tests. At 16 weeks, bone mineral density, cortical load, yield, and stiffness as well as cancellous bone failure stress, yield stress, and modulus were significantly lower (p < 0.02) for immobilized limbs than control limbs. Immobilized limb cancellous bone mechanical properties were 28%-74% of control values, and cortical bone mechanical properties were 71%-98% of control values. After 32 weeks of remobilization, cortical and cancellous bone mechanical properties were not different from control values except that cortical bone failure stress and modulus were significantly higher (p < 0.01) between remobilized and control limbs. In summary, 16 weeks of forelimb immobilization was associated with significantly lower mechanical properties, and with greater differences in cancellous than cortical bone properties. Mechanical properties were not different from control values after 32 weeks of recovery that included 16 weeks of treadmill exercise.     

Vitamin C elevates red blood cell glutathione in healthy adults

Pre-transplant nephrectomy was done in a 25-year-old man for calculous pyelonephritis using a retroperitoneal laparoscopic approach with a newly devised ligature applicator-dissector- kidney retractor.     

Childhood fibrous dysplasia presenting as blindness: a skull base approach for resection and immediate reconstruction

Fibrous dysplasia is an abnormal fibroosseous process of bone of unknown cause. The incidence of skull involvement varies, painless enlargement being the most common presenting symptom. Change in vision is a rare but recognized finding. We report a 3-year-old boy with extreme fibrous dysplasia involving the skull base, who presented with blindness. He underwent exposure osteotomies of the frontal bones and orbits to provide access for skull base tumor removal. The orbital roofs were reconstructed with microplate-fixed cranial grafts. One and one half years after tumor excision followed by immediate reconstruction, the boy retains facial symmetry, and his ocular function has not deteriorated.     

Multicast routing and bandwidth dimensioning in overlay networks

Multicast services can be provided either as a basic network service or as an application-layer service. Higher level multicast implementations often provide more sophisticated features and can provide multicast services at places where no network layer support is available. Overlay multicast networks offer an intermediate option, potentially combining the flexibility and advanced features of application layer multicast with the greater efficiency of network layer multicast. In this paper, we introduce the multicast routing problem specific to the overlay network environment and the related capacity assignment problem for overlay network planning. Our main contributions are the design of several routing algorithms that optimize the end-to-end delay and the interface bandwidth usage at the multicast service nodes within the overlay network. The interface bandwidth is typically a key resource for an overlay network provider, and needs to be carefully managed in order to maximize the number of users that can be served. Through simulations, we evaluate the performance of these algorithms under various traffic conditions and on various network topologies. The results show that our approach is cost-effective and robust under traffic variations.     

Logical foundations of nonmonotonic reasoning

In this paper we shall review the main appraches to nonmonotonic reasoning which we classify from the perspective of their underlying logical settings as classical, intuitionistic, three-valued/partial models, and conditional. We shall be placing special emphasis on some of the prominent approaches. We shall also give hints on potential future directions and emphasize that more theoretical work is still needed before a move to application is made.