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.     

Evaluating hyperspectral imaging of wetland vegetation as a tool for detecting estuarine nutrient enrichment

Nutrient enrichment and eutrophication are major concerns in many estuarine and wetland ecosystems, and the need is urgent for fast, efficient, and synoptic ways to detect and monitor nutrients in wetlands and other coastal systems across multiple spatial and temporal scales. We integrated three approaches in a multi-disciplinary evaluation of the potential for using hyperspectral imaging as a tool to assess nutrient enrichment and vegetation responses in tidal wetlands. For hyperspectral imaging to be an effective tool, spectral signatures must vary in ways correlated with water nutrient content either directly, or indirectly via such proxies as vegetation responses to elevated nitrogen. Working in Elkhorn Slough, central California, where intensive farming practices generate considerable runoff of fertilizers and pesticides, we looked first for long- and short-term trends among temporally ephemeral point data for nutrients and other water quality characters collected monthly at 18 water sampling stations since 1988. Second, we assessed responses of the dominant wetland plant, Salicornia virginica (common pickleweed) to two fertilizer regimes in 0.25 m² experimental plots, and measured changes in tissue composition (C, H, N), biomass, and spectral responses at leaf and at canopy scales. Third, we used HyMap hyperspectral imagery (126 bands; 15–19 nm spectral resolution; 2.5 m spatial resolution) for a synoptic assessment of the entire wetland ecosystem of Elkhorn Slough. We mapped monospecific Salicornia patches (～ 56–500 m²) on the ground adjacent to the 18 regular water sampling sites, and then located these patches in the hyperspectral imagery to correlate long-term responses of larger patches to water nutrient regimes. These were used as standards for correlating plant canopy spectral responses with nitrogen variation described by the water sampling program. There were consistent positive relationships between nitrogen levels and plant responses in both the field experiment and the landscape analyses. Two spectral indices, the Photochemical Reflectance Index (PRI) and Derivative Chlorophyll Index (DCI), were correlated significantly with water nutrients. We conclude that hyperspectral imagery can be used to detect nutrient enrichment across three spatial and at least two temporal scales, and suggest that more quantitative information could be extracted with further research and a greater understanding of physiological and physical mechanisms linking water chemistry, plant properties and spectral imaging characteristics.     

Sea-air flux of CO2 in the Caribbean Sea estimated using in situ and remote sensing data

Empirical relationships between sea surface carbon dioxide fugacity (fCO₂^sw) and sea surface temperature (SST) were applied to datasets of remotely sensed SST to create fCO₂^sw fields in the Caribbean Sea. SST datasets from different sensors were used, as well as the SST fields created by optimum interpolation of bias corrected AVHRR data. Empirical relationships were derived using shipboard fCO₂^sw data, in situ SST data, and SST data from the remote sensing platforms. The results show that the application of a relationship based on shipboard SST data, on fields of remotely sensed SST yields biased fCO₂^sw values. This bias is reduced if the fCO₂^sw-SST relationships are derived using the same SST data that are used to create the SST fields. The fCO₂^sw fields found to best reproduce observed fCO₂^sw are used in combination with wind speed data from QuikSCAT to create weekly maps of the sea-air CO₂ flux in the Caribbean Sea in 2002. The region to the SW of Cuba was a source of CO₂ to the atmosphere throughout 2002, and the region to the NE was a sink during winter and spring and a source during summer and fall. The net uptake of CO₂ in the region was doubled when potential skin layer effects on fCO₂^sw were taken into account.     

Thoracic and abdominal SPECT-CT image fusion without external markers in endocrine carcinomas. The Group of Thyroid Tumoral Pathology of Champagne-Ardenne

Superimposition of SPECT and computed tomography (CT) slices from the thoracoabdominal region was achieved without the use of external markers for 14 studies in 13 patients with endocrine carcinoma. Technical feasibility and clinical validation of this retrospective fusion method were assessed. METHODS: Patients had a history of thyroid cancer or of carcinoid tumor. To detect tumor sites, CT scan and dual-isotope tomoscintigraphy were performed, with 99mTc-hydroxymethylene diphosphonate for bone scintigraphy and with 111In-pentetreotide, 131I or 131I-metaiodobenzylguanidine for tumor scintigraphy (TS). A superimposition method previously developed for the pelvic region was adapted to the nonrigid thoraco-abdominal region. CT-bone scintigraphy and CT-TS superimposed images were obtained. Clinical validation of the information obtained from the superimposed images was obtained from surgery or follow-up imaging studies performed after clinical evolution of the disease process. RESULTS: Reliable and reproducible registration was achieved in all patients. CT-TS superimposed images produced accurate localization of abnormal TS foci. Accuracy was limited primarily by variable relative displacements of the thoracoabdominal organs. For 10 sites in 8 patients, localization and/or characterization obtained from CT-TS images was confirmed by a reference technique. Superimposition enabled the localization of tumor sites that otherwise could not have been suspected from CT alone and allowed the characterization of CT suspicious masses and the confirmation of CT positive sites. Nonspecific tumor TS uptake sites were also localized. CONCLUSION: With standard CT and dual-isotope SPECT acquisitions, SPECT-CT fusion is feasible in the thoracoabdominal region without the use of external markers. Fused images were validated in 8 patients for 10 sites. The use of this technique could probably improve the management and care of patients with endocrine carcinoma.