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Dephospho-coenzymeA kinase catalyzes the final step in CoA biosynthesis, the phosphorylation of the 3′-hydroxyl group of ribose using ATP as a phosphate donor. The protein from Haemophilus influenzae was cloned and expressed, and its crystal structure was determined at 2.0-A resolution in complex with ATP. The protein molecule consists of three domains: the canonical nucleotide-binding domain with a five-stranded parallel β-sheet, the substrate-binding α-helical domain, and the lid domain formed by a pair of α-helices. The overall topology of the protein resembles the structures of nucleotide kinases. ATP binds in the P-loop in a manner observed in other kinases. The CoA-binding site is located at the interface of all three domains. The double-pocket structure of the substrate-binding site is unusual for nucleotide kinases. Amino acid residues implicated in substrate binding and catalysis have been identified. The structure analysis suggests large domain movements during the catalytic cycle.

We have previously reported on the use of an open flow helium cryostat in macromolecular crystallographic data collection. This device, developed at the University of Toledo, was used at the APS IMCA beamline to test He as a cryogen and to compare data collected at He temperature to that collected at N 2 temperature. Data were collected on matched crystals of the nucleosome core particle (NCP), fumarylacetoacetate hydrolase (FAH), and the murine monoclonal antibody BV0401 (BV), and other crystals using an Oxford Cryosystems Cryostream and the Toledo Helium Cryostat. In these experiments, crystal lifetime was enhanced by using He as a cryogen. At the end of data collection with NCP, crystals cooled by N 2 were blackened; crystals cooled by He remained clear. A crystal of Mcg light chain dimer (Mcg) had undiminished diffraction in He after 90 min of exposure in the same position. Data improvement is best illustrated in the diffraction data from BV. These data showed an improvement in resolution (1.75–1.48 A) with a significant enhancement of the intensity for weak data. Data from the NCP showed a similar enhancement in intensity. However, some diffraction data exhibited split reflections at He temperature rendering processing of the data more problematic. One possible explanation for these results is a phase change occurring between 100 K and the 30 K of the helium cold stream. Planned work will characterize this phenomenon and extend the experiments to other crystal systems.

Benzo-fused model compounds 21a and 21b, resembling in structure the marine metabolite ptilomycalin A, were prepared and were shown to display significant activity against a series of cancer cell lines and to also possess a significant activity against the DNA polymerase activity of the reverse transcriptase of human immunodeficiency virus type 1 (HIV-1 RT). © 2001 Elsevier Science Ltd. All rights reserved. Ptilomycalin A 1 and related natural products have been the subject of considerable synthetic interest owing to their high levels of diverse biological activity. This includes cytotoxicity towards several cancer cell lines as well as antifungal and antiviral activities. 1 Despite this range of activity, only one report of the synthesis of a structural analogue of ptilomycalin A has been reported, by Hart and Grillot, which detailed the prepa- ration of the racemic bicyclic guanidine 2 2 (Scheme 1). As can be seen, ptilomycalin A can be considered as four separate components, a guanidine-containing poly- cyclic core, an ester linkage to an -hydroxyacid spacer unit and a terminal spermidine residue. In common

The observation of light metal ions in nucleic acids crystals is generally a fortuitous event. Sodium ions in particular are notoriously difficult to detect because their X-ray scattering contributions are virtually identical to those of water and Na+…O distances are only slightly shorter than strong hydrogen bonds between well-ordered water molecules. We demonstrate here that replacement of Na+ by K+, Rb+ or Cs+ and precise measurements of anomalous differences in intensities provide a particularly sensitive method for detecting alkali metal ion-binding sites in nucleic acid crystals. Not only can alkali metal ions be readily located in such structures, but the presence of Rb+ or Cs+ also allows structure determination by the single wavelength anomalous diffraction technique. Besides allowing identification of high occupancy binding sites, the combination of high resolution and anomalous diffraction data established here can also pinpoint binding sites that feature only partial occupancy. Conversely, high resolution of the data alone does not necessarily allow differentiation between water and partially ordered metal ions, as demonstrated with the crystal structure of a DNA duplex determined to a resolution of 0.6 Å.

The crystal structure of YecO from Haemophilus influenzae (HI0319), a protein annotated in the sequence databases as hypothetical, and that has not been assigned a function, has been determined at 2.2-A resolution. The structure reveals a fold typical of S-adenosyl-L-methionine-dependent (AdoMet) methyltransferase enzymes. Moreover, a processed cofactor, S-adenosyl-L-homocysteine (AdoHcy), is bound to the enzyme, further confirming the biochemical function of HI0319 and its sequence family members. An active site arginine, shielded from bulk solvent, interacts with an anion, possibly a chloride ion, which in turn interacts with the sulfur atom of AdoHcy. The AdoHcy and nearby protein residues delineate a small solvent-excluded substrate binding cavity of 162 A(3) in volume. The environment surrounding the cavity indicates that the substrate molecule contains a hydrophobic moiety and an anionic group. Many of the residues that define the cavity are invariant in the HI0319 sequence family but are not conserved in other methyltransferases. Therefore, the substrate specificity of YecO enzymes is unique and differs from the substrate specificity of all other methyltransferases sequenced to date. Examination of the Enzyme Commission list of methyltransferases prompted a manual inspection of 10 possible substrates using computer graphics and suggested that the ortho-substituted benzoic acids fit best in the active site.

The enzyme chorismate lyase (CL) catalyzes the removal of pyruvate from chorismate to produce 4-hydroxy benzoate (4HB) for the ubiquinone pathway. In Escherichia coli, CL is monomeric, with 164 residues. We have determined the structure of the CL product complex by crystallographic heavy-atom methods and report the structure at 1.4-A resolution for a fully active double Cys-to-Ser mutant and at 2.0-A resolution for the wild-type. The fold involves a 6-stranded antiparallel β-sheet with no spanning helices and novel connectivity. The product is bound internally, adjacent to the sheet, with its polar groups coordinated by two main-chain amides and by the buried side-chains of Arg 76 and Glu 155. The 4HB is completely sequestered from solvent in a largely hydrophobic environment behind two helix–turn–helix loops. The extensive product binding that is observed is consistent with biochemical measurements of slow product release and 10-fold stronger binding of product than substrate. Substrate binding and kinetically rate-limiting product release apparently require the rearrangement of these active-site-covering loops. Implications for the biological function of the high product binding are considered in light of the unique cellular role of 4HB, which is produced by cytoplasmic CL but is used by the membrane-bound enzyme 4HB octaprenyltransferase. Proteins 2001;44:304–311. © 2001 Wiley-Liss, Inc.

The crystal structure of the Bacillus subtilis chorismate mutase, an enzyme of the aromatic amino acids biosynthetic pathway, was determined to 1.30 A resolution. The structure of the homotrimer was determined by molecular replacement using orthorhombic crystals of space group P2(1)2(1)2(1) with unit-cell parameters a = 52.2, b = 83. 8, c = 86.0 A. The ABC trimer of the monoclinic crystal structure [Chook et al. (1994), J. Mol. Biol. 240, 476-500] was used as the starting model. The final coordinates are composed of three complete polypeptide chains of 127 amino-acid residues. In addition, there are nine sulfate ions, five glycerol molecules and 424 water molecules clearly visible in the structure. This structure was refined with aniosotropic temperature factors, has excellent geometry and a crystallographic R factor of 0.169 with an R(free) of 0.236. The three active sites of the macromolecule are at the subunit interfaces, with residues from two subunits contributing to each site. This orthorhombic crystal form was grown using ammonium sulfate as the precipitant; glycerol was used as a cryoprotectant during data collection. A glycerol molecule and sulfate ion in each of the active sites was found mimicking a transition-state analog. In this structure, the C-terminal tails of the subunits of the trimer are hydrogen bonded to residues of the active site of neighboring trimers in the crystal and thus cross-link the molecules in the crystal lattice.

This paper describes the navigation system used by J Edgar, a small vision-guided robot that roams the corridors at the University of Melbourne. Given a model of the environment, J Edgar is able to find its way to any given location, even when its initial location is completely unknown. To do this, the robot makes use of a probabilistic localisation technique, in which the robot maintains a probability distribution over the space of all possible robot locations. Given this distribution, the robot then applies a particular navigation strategy, whose aim is to ensure that the robot not only reaches the goal, but knows that it has reached the goal.

Carbamoyl phosphate synthetase from Escherichia coli catalyzes the production of carbamoyl phosphate from two molecules of Mg 2+ ATP, one molecule of bicarbonate, and one molecule of glutamine. The enzyme consists of two polypeptide chains referred to as the large and small subunits. While the large subunit provides the active sites responsible for the binding of nucleotides and other effector ligands, the small subunit contains those amino acid residues that catalyze the hydrolysis of glutamine to glutamate and ammonia. From both amino acid sequence analyses and structural studies it is now known that the small subunit belongs to the class I amidotransferase family of enzymes. Numerous biochemical studies have suggested that the reaction mechanism of the small subunit proceeds through the formation of the glutamyl thioester intermediate and that both Cys 269 and His 353 are critical for catalysis. Here we describe the X-ray crystallographic structure of carbamoyl phosphate synthetase from E. coli in which His 353 has been replaced with an asparagine residue. Crystals employed in the investigation were grown in the presence of glutamine, and the model has been refined to a crystallographic R-factor of 19.1% for all measured X-ray data from 30 to 1.8 A resolution. The active site of the small subunit clearly contains a covalently bound thioester intermediate at Cys 269, and indeed, this investigation provides the first direct structural observation of an enzyme intermediate in the amidotransferase family.

The recent rise in systemic fungal infections has created a need for the development of new antifungal agents. As part of an effort to provide therapeutically effective inhibitors of fungal dihydrofolate reductase (DHFR), we have cloned, expressed, purified, crystallized, and determined the three-dimensional structure ofCandida albicans DHFR. The 192-residue enzyme, which was expressed in Escherichia coli and purified by methotrexate affinity and cation exchange chromatography, was 27% identical to human DHFR. Crystals of C. albicans DHFR were grown as the holoenzyme complex and as a ternary complex containing a pyrroloquinazoline inhibitor. Both complexes crystallized with two molecules in the asymmetric unit in space group P21. The final structures had R-factors of 0.199 at 1.85-A resolution and 0.155 at 1.60-A resolution, respectively. The enzyme fold was similar to that of bacterial and vertebrate DHFR, and the binding of a nonselective diaminopyrroloquinazoline inhibitor and the interactions of NADPH with protein were typical of ligand binding to other DHFRs. However, the width of the active site cleft of C. albicans DHFR was significantly larger than that of the human enzyme, providing a basis for the design of potentially selective inhibitors.

Positron emission tomography (PET) is currently the premiere molecular imaging technique for the detection and staging of cancer in vivo. Owing to the desirable nuclear properties of the short- lived positron emitter 18F, and the remarkable targeting potential of many peptides, proteins and nucleic acid- based compounds, the efficient radiolabelling of these sensitive biomolecules with 18F remains a fundamental objective in nuclear diagnostic medicine. Antisense imaging remains a promising tool for the identification and treatment of genetic diseases, particularly cancer. Herein, we expand the utility of the 2-[18F]fluoropyridine- bearing labelling agent [18F]FPy5yne to include nucleic acid- based probes for PET imaging. We describe the conjugation of [18F]FPy5yne by way of a copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction to a 5'-azide-modified antisense oligodeoxyribonucleotide, which was first prepared from a 5'-aminohexyl-modified DNA 20mer. This work also describes protocols suitable for the coupling of [18F]FPy5yne and mini-PEGylated analog PEG-[18F]FPyKYNE to azide- modified peptide receptor ligands derived from bombesin and neuropeptide Y. These 18F- labelled targeting vectors were assayed by collaborators for the detection of prostate, breast and brain cancer in mouse models by way of PET imaging and ex vivo autoradiography. Finally, a new approach for the 18F- labelling of biological molecules is introduced and investigated. Sulfonyl [18F]fluorides can be prepared under aqueous conditions and at room temperature, yet they have not yet been assayed as a potential means to 18F- label biomarkers. A general route was developed for the synthesis of bifunctional arylsulfonyl [18F]fluorides from their sulfonyl chloride precursors in 1:1 mixtures of MeCN, THF, or t-BuOH and Cs[18F]F/Cs2CO3 over 15 minutes at room temperature. In most cases, pyridine could be used to selectively degrade the precursor without significantly affecting observed yields. As proof-of-principle, 3-formylmesitylenesulfonyl [18F]fluoride was synthesized in excellent preparative yields and used to radiolabel an oxyamino-modified bombesin(6-14) analog. The 18F bioconjugate showed signs of both defluorination and modification in mouse serum.

Background Walking to school is a way to increase daily physical activity; however the risk of injury must also be considered so that walking does not lead to an increase in pedestrian injuries. Risk factors associated with the environment around schools with high child pedestrian motor vehicle collision (PMVC) rates were examined. Methods Child PMVCs from 2000–2013, ages 4–12 years, were mapped within elementary school attendance boundaries in Toronto, Canada. Case and control schools were defined as those with the highest and lowest quartile of PMVC rates, calculated using census data. Potential risk factors included built and non-built environment variables obtained from municipal data sources as well as via direct observational counts done in the spring, 2015, to measure the proportion of children walking to school. Logistic regression was used to compare case versus control schools stratified by geographic location (downtown vs inner suburbs). Results The mean PMVC rate in case schools (n = 50) was 13.4/10,000/year and in controls (n = 50) was 1.75/10,000/year. Walking was not associated with high PMVC rates after adjustment for the built environment and school social disadvantage. Overall, lower residential (OR 0.56, 95% CI: 0.37, 0.86) and higher one-way street densities (OR 4.00, 95% CI: 1.76, 9.08), school crossing guards (OR 3.65, 95% CI: 1.10, 12.20) and higher social disadvantage (OR 1.37, 95% CI: 1.11, 1.70) were associated with high PMVC schools, Similar associations of high PMVC schools with built environment features were found in the inner suburbs; however, there was a stronger association with school social disadvantage downtown. Conclusions Walking to school was unrelated to high PMVC rates after controlling for the built environment. The built environment and school disadvantage were associated with higher PMVC rates with possible differences by geographic location.

Background Dangerous drop-off of student passengers and pedestrian behaviours near schools have not been well described despite the potential for pedestrian motor vehicle collisions (PMVC). Safe environments are required for children around schools. Passenger drop-off and child pedestrian behaviours are described by occurrence of child PMVC and built environment (BE) features in Toronto, Canada. Methods Dangerous passenger drop-off and pedestrian behaviours observations were done in 2015. Child PMVCs from 2000–2013, age 4–12 years, were mapped near 100 schools. Case schools had > 1 collision and control schools had 0. Dangerous driving/parking, unsafe pedestrian crossings, distracted walking and disobeying crossing controls/guards were compared using chi-square statistics by case/control status or BE features: downtown/inner suburbs, school crossing guards and designated car drop-off areas. Results Dangerous drop-off and/or pedestrian behaviours occurred at over 92% of schools. A greater proportion of case schools had children crossing at uncontrolled midblocks (97% vs. 78% controls). A greater proportion of inner suburb schools versus downtown had cars double-parked (52% vs. 28%) and parked blocking crossing controls (25% vs. 4%). A smaller proportion of schools with crossing guards versus without had cars parked blocking crossing controls (10% vs. 25% without) and drivers texting (5% vs. 22%). A smaller proportion of schools with designated drop-off areas versus without, had cars reversing dangerously (76% vs. 55%), parked blocking crossing controls (31% vs. 10%), children crossing at uncontrolled midblocks (78% vs. 95%) and between parked cars (48% vs. 76%). Conclusions Dangerous student drop-off and pedestrian behaviours were pervasive at schools. Occurrence of behaviours was related to the BE; particularly designated car-drop off areas. Adaptations to the BE near schools may defer dangerous behaviours and provide a safer child pedestrian environment.

Labor internationalism has historically developed in response to the increasing globalization of capital, but economic factors in themselves do not provide a sufficient explanation for the relative success of international solidarity in specific geographical and historical contexts. In the past, movements of international solidarity have tended to coalesce where inclusive class-based mobilizations in individual countries have polarized local class conflicts in ways that suggest a convergence of working-class interests on the international plane. As an emergent strategy of economistic labor organizations in various sectors of the capitalist world economy, however, internationalism may in fact promote the politicization of economic trade union struggles and thus catalyze broader levels of class solidarity and unity on the national and local planes.

The crystal complex of fluorescein bound to the high-affinity anti-fluorescein 4-4-20 Fab (Ka = 10(10) M-1 at 2 degrees C) has been determined at 1.85 A. Isomorphous crystals of two isoelectric forms (pI = 7.5 and 7.9) of the anti-fluorescein 4-4-20 Fab, an IgG2A [Gibson et al. (1988) Proteins: Struct. Funct. Genet., 3, 155-160], have been grown. Both complexes crystallize with one molecule in the asymmetric unit in space group P1, with a = 42.75 A, b = 43.87 A, c = 58.17 A, alpha = 95.15 degrees, beta = 86.85 degrees and gamma = 98.01 degrees. The final structure has an R value of 0.188 at 1.85 A resolution. Interactions between bound fluorescein, the complementarity-determining regions (CDRs) of the Fab and the active-site mutants of the 4-4-20 single-chain Fv will be discussed. Differences were found between the structure reported here and the previously reported 2.7 A 4-4-20 Fab structure [Herron et al. (1989) Proteins: Struct. Funct. Genet., 5, 271-280]. Our structure determination was based on 26,328 unique reflections--four times the amount of data used in the previous report. Differences in the two structures could be explained by differences in interpreting the electron density maps at the various resolutions. The r.m.s. deviations between the variable and constant domains of the two structures were 0.77 and 1.54 A, respectively. Four regions of the light chain and four regions of the heavy chain had r.m.s. backbone deviations of > 4 A. The most significant of these was the conformation of the light chain CDR 1.

Ten percent of the 250,000 proximal femur fractures that occur in the United States each year are malreduced into a varus position after treatment. Currently, there is no cephalomedullary nail available that allows the physician to dynamically change the lag-screw-to-nail angle. The Variable Angle Nail (VAN) was designed to allow movement of the lag screw relative to the shaft of the nail. This study compared the characteristics of the VAN to the Gamma 3 nail via finite element analysis (FEA) in stiffness and fatigue. The results of the FEA model with the same loading parameters showed the Gamma 3 and the VAN with lag-screw-to-nail angle of 120° to have essentially the same stiffness values ranging from 350 to 382 N/mm. The VAN with lag-screw-to-nail angles of 120°, 130°, and 140° should be able to withstand more than 1,000,000 cycles from 1,400 N to 1,500 N loading of the tip of the lag screw. The Gamma 3 should be able to last more than 1,000,000 cycles at 1,400 N. In summary, the VAN is superior or equivalent in stiffness and fatigue when compared to the Gamma 3 using FEA.

Fourth-year medical students (MS4s) participate in clerkships to familiarize themselves with the field of radiation oncology and to evaluate and audition for residency programs. However, MS4s’ perceptions of their radiation oncology clerkship experiences have never been formally evaluated. This study aimed to characterize the MS4 perception of the radiation oncology clerkship experience from the current nationwide pool of applicants.

Analyses some of the reasons for fatigue and stress among drivers. Contributors to fatigue can be the distance travelled, or sleeping disorders. Contributors to driver stress can include worry and emotional stress, related to work or to life events; or road rage. Identifies ways in which stress, fatigue and road rage can be reduced or avoided. Provides advice for fleet managers on making driving safer.

Saccharomyces cerevisiae alcohol dehydrogenase I crystallized as trigonal plates using 20% 2-propanol and 20% PEG 4000 in 0.1 M sodium citrate buffer pH 5.6 in the presence of 1 mM NAD(+). The crystals diffract to 3.0 A resolution and belong to the trigonal space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 146.3, c = 68.1 A, alpha = beta = 90, gamma = 120 degrees. X-ray data were collected from frozen crystals at the 17-ID beamline of the Advanced Photon Source. A Zn fluorescence scan of the crystal produced a peak at 9671.6 eV, suggesting the existence of Zn ions in the crystal.

On 1 February 2011 the Kepler Mission released data for 156,453 stars observed from the beginning of the science observations on 2 May through 16 September 2009. There are 1235 planetary candidates with transit like signatures detected in this period. These are associated with 997 host stars. Distributions of the characteristics of the planetary candidates are separated into five class-sizes; 68 candidates of approximately Earth-size (radius < 1.25 Earth radii), 288 super-Earth size (1.25 Earth radii < radius < 2 Earth radii), 662 Neptune-size (2 Earth radii < radius < 6 Earth radii), 165 Jupiter-size (6 Earth radii < radius < 15 Earth radii), and 19 up to twice the size of Jupiter (15 Earth radii < radius < 22 Earth radii). In the temperature range appropriate for the habitable zone, 54 candidates are found with sizes ranging from Earth-size to larger than that of Jupiter. Five are less than twice the size of the Earth. Over 74% of the planetary candidates are smaller than Neptune. The observed number versus size distribution of planetary candidates increases to a peak at two to three times Earth-size and then declines inversely proportional to area of the candidate. Our current best estimates of the intrinsic frequencies of planetary candidates, after correcting for geometric and sensitivity biases, are 6% for Earth-size candidates, 7% for super-Earth size candidates, 17% for Neptune-size candidates, and 4% for Jupiter-size candidates. Multi-candidate, transiting systems are frequent; 17% of the host stars have multi-candidate systems, and 33.9% of all the candidates are part of multi-candidate systems., 106 pages, 15 figures, contains tables of candidates. Submitted to ApJ

The EORTC/NCIC 22981/26981 study demonstrated an improvement in median overall survival (OS) from 12.1 to 14.6 months in patients with glioblastoma (GBM) who received temozolomide with post-operative radiotherapy (RT). The current study was performed to determine if those results translated into a survival benefit in a population-based cohort. Patients diagnosed between 2000 and 2006 with a GBM who underwent surgery and post-operative RT were selected from the Surveillance, Epidemiology and End Results database. Patients were grouped into time periods: 2000-2001, 2002-2003, 2004 and 2005-2006 (which represented those treated after the EORTC/NCIC trial presentation in 2004). Relative survival (RS) was estimated by the Kaplan-Meier method, and Cox multivariable regression modeling was used to estimate proportional hazard ratios (HR). Over time, there was improvement in the median and 2-year RS of 12 months and 15% for 2000-2001, 13 months and 19% for 2002-2003, 14 months and 24% for 2004, and 15 months and 26% for 2005-2006 (P < 0.0001 compared to 2000-2001 and 2002-2003; P = 0.07 compared to 2004). The estimated adjusted HR showed that patients diagnosed in 2005-2006 had significantly improved survival when compared to patients diagnosed in 2000-2001 (HR = 0.648, 95% CI 0.604-0.696). The median and 2 year RS of 15 months and 26% in 2005-2006 was similar to the median and 2 year OS of 14.6 months and 26% seen in the EORTC/NCIC phase III study. These results are encouraging and suggest that the current treatment of glioblastoma nationwide is now associated with an improved survival compared to previous time cohorts.

♦ All children who complain of knee symptoms must be assessed for ipsilateral hip and spine pathology♦ Congenital or persistent lateral dislocation of the patella and obligatory dislocation of the patella have two different clinical presentations: surgical treatment (if required) is often complex♦ The natural history of stable osteochondritis dissecans lesions is generally favourable in a child with open physes.

Single chain Fv (sFv) proteins consist of the variable heavy chain (VH) and variable light chain (VL) domains of an antibody, covalently joined by an engineered polypeptide linker. We report the crystallization of single-chain Fv's with specificities for fluorescein (4-4-20 sFv) and the TAG-72 pancarcinoma glycoprotein antigen (CC49 sFv). Concentration of these proteins, preliminary to crystallization, results in a monomer-multimer equilibrium, causing aggregation which interferes with crystallization. Aggregation has been observed to depend primarily on an intact linker between VL and VH domains, although other factors are likely to modulate this phenomenon as well, including the specific identity of Fv and ligand, presence or absence of the ligand, linker length and possibly sequence. We have found two methods to overcome sFv aggregation, both of which yield X-ray diffraction quality crystals. The first, discovered serendipitously, is by introducing a proteolytic clip into the linker region (effectively yielding an Fv fragment). The second is the purification of the sFv dimer form, with linker regions intact, from an equilibrium mixture of aggregates. The sFv molecular association in a dimer is believed to be unusual in that each VL/VH interface may not be formed by the two linker connected VL and VH domains, but rather by interaction of VL and VH domains from two distinct sFv monomers. Structure determination of the CC49 sFv dimer, with the 14-residue linker designated 212, is underway to test this model. Increasing linker length, to relieve steric strain on the monomer, and inclusion of the appropriate antigen, to slow transitions between monomeric and multimeric forms, may prove valuable strategies with sFv proteins less amenable to crystallization.

Low-bandwidth video has a place in health service delivery. Videoconferencing systems commonly used in telehealth have a high capital cost, take days or weeks to install at specific locations, and have high communications charges. A range of circumstances can be envisaged where video may be of benefit but would not justify the cost of large systems. There are projects in community nursing, home dialysis and post-acute respiratory care where various low-bandwidth video technologies have been added as a supplement to the humble telephone call. It can be expected that at least some of these projects will be able to demonstrate tangible improvements in health outcomes, such as decreased readmission rates, reduced acute episodes of a chronic illness and improved health status from the additional social support.

The End-Stage Renal Disease Network 5 sought to improve the influenza vaccination rate for the period September 1, 2008, to January 31, 2009, through an awareness campaign, coupled with primary data collection in the form of a tracking tool prepopulated with patient names. The latter served as a reminder to staff to determine the immunization status of patients and offer the influenza vaccination, as appropriate. Targets for the intervention were all facilities and their prevalent hemodialysis and peritoneal dialysis patients, with the exclusion of military treatment centers, Veterans Health Administration hospitals, and prisons. The majority of eligible network facilities (86.9%) participated in the project to achieve an overall adult influenza vaccination rate of 82.6% (95% confidence interval = 82.1%, 83.2%), greatly exceeding the project goal of 64.5% and representing substantial progress toward the 2010 goal of 90%. The initiative is reported here using the Standards for Quality Improvement Reporting Excellence (SQUIRE).

Glycocyamine kinase (GK), a member of the phosphagen kinase family, catalyzes the Mg(2+)-dependent reversible phosphoryl group transfer of the N-phosphoryl group of phosphoglycocyamine to ADP to yield glycocyamine and ATP. This reaction helps to maintain the energy homeostasis of the cell in some multicelullar organisms that encounter high and variable energy turnover. GK from the marine worm Namalycastis sp. is heterodimeric, with two homologous polypeptide chains, alpha and beta, derived from a common pre-mRNA by mutually exclusive N-terminal alternative exons. The N-terminal exon of GKbeta encodes a peptide that is different in sequence and is 16 amino acids longer than that encoded by the N-terminal exon of GKalpha. The crystal structures of recombinant GKalphabeta and GKbetabeta from Namalycastis sp. were determined at 2.6 and 2.4 A resolution, respectively. In addition, the structure of the GKbetabeta was determined at 2.3 A resolution in complex with a transition state analogue, Mg(2+)-ADP-NO(3)(-)-glycocyamine. Consistent with the sequence homology, the GK subunits adopt the same overall fold as that of other phosphagen kinases of known structure (the homodimeric creatine kinase (CK) and the monomeric arginine kinase (AK)). As with CK, the GK N-termini mediate the dimer interface. In both heterodimeric and homodimeric GK forms, the conformations of the two N-termini are asymmetric, and the asymmetry is different than that reported previously for the homodimeric CKs from several organisms. The entire polypeptide chains of GKalphabeta are structurally defined, and the longer N-terminus of the beta subunit is anchored at the dimer interface. In GKbetabeta the 24 N-terminal residues of one subunit and 11 N-terminal residues of the second subunit are disordered. This observation is consistent with a proposal that the GKalphabeta amino acids involved in the interface formation were optimized once a heterodimer emerged as the physiological form of the enzyme. As a consequence, the homodimer interface (either solely alpha or solely beta chains) has been corrupted. In the unbound state, GK exhibits an open conformation analogous to that observed with ligand-free CK or AK. Upon binding the transition state analogue, both subunits of GK undergo the same closure motion that clasps the transition state analogue, in contrast to the transition state analogue complexes of CK, where the corresponding transition state analogue occupies only one subunit, which undergoes domain closure. The active site environments of the GK, CK, and AK at the bound states reveal the structural determinants of substrate specificity. Despite the equivalent binding in both active sites of the GK dimer, the conformational asymmetry of the N-termini is retained. Thus, the coupling between the structural asymmetry and negative cooperativity previously proposed for CK is not supported in the case of GK.

Doppler observations from Keck Observatory reveal the presence of a planet with M sin i of 2.1 M_(Jup) orbiting the M3V star HIP 79431. This is the sixth giant planet to be detected in Doppler surveys of M dwarfs and it is one of the most massive planets discovered around an M dwarf star. The planet has an orbital period of 111.7 days and an orbital eccentricity of 0.29. The host star is metal rich, with an estimated [Fe/H] = +0.4. This is the first planet to emerge from our new survey of 1600 M-to-K dwarf stars.

The structure of the B2 immunoglobulin-binding domain of streptococcal protein G has been determined at 1.67-A resolution using a combination of single isomorphous replacement (SIR) phasing and manual fitting of the coordinates of the NMR structure of B 1 domain of streptococcal protein G (Gronenborn, A. M., et al. (1991) Science 253, 657-6611, The final R value was 0.191 for data between 8.0 and 1.67 A. The structure described here has 13 residues preceding the 57-residue Ig-binding domain and 13 additional residues following it, for a total of 83 residues. The 57-residue binding domain is well-determined in the structure, having an average B factor of 18.0. Only residues 8-77 could be located in the electron density maps, with the ends of the structure fading into disorder. Like the B1 domain, the B2 domain consists of four 6-strands and a single helix lying diagonally across the s these globular domains are connected by extended linkers (Fahnestocket al., 1990). Protein G offersseveraladvantages over protein A for the binding of IgG immunoglobins. Protein G binds to all subclasses of human IgG, whereas protein A does not bind the IgG3 subclass (Reis et al., 1984). Protein G binds a number of animal IgGs to which protein A binds only weakly. Protein G has a number of useful applications, including the treatment of diseases and the detection and purification of IgG's. Protein G could be used in the removal of IgGs from plasma, using an extracorporeal column (Tenan et al., 1981). The detection of IgG's is important in screening for hybridoma clones, in quantitative studies of immune response in animals, and in detection of antigens by competition-binding

This article proposes an algorithm to automatically learn useful transformations of data to improve accuracy in supervised classification tasks. These transformations take the form of a mixture of base transformations and are learned by maximizing the kernel alignment criterion. Because the proposed optimization is nonconvex, a semidefinite relaxation is derived to find an approximate global solution. This new convex algorithm learns kernels made up of a matrix mixture of transformations. This formulation yields a simpler optimization while achieving comparable or improved accuracies to previous transformation learning algorithms based on maximizing the margin. Remarkably, the new optimization problem does not slow down with the availability of additional data allowing it to scale to large datasets. One application of this method is learning monotonic transformations constructed from a base set of truncated ramp functions. These monotonic transformations permit a nonlinear filtering of the input to the classifier. The effectiveness of the method is demonstrated on synthetic data, text data and image data.

Frontier Exploration Licence 7/97 is located 115km offshore north-west Ireland and directly overlies the Erris Ridge. The ridge is an enigmatic, narrow, segmented, NE-SW orientated structural high which lies between the Erris Basin to the east and the Irish Rockall Trough to the west and which, to date, remains un-drilled. Seismic data quality across the Erris Ridge is typically poor and correlation with the sparse offset well database is challenging. Zones of poor seismic imaging are associated with combinations of complex shallow volcaniclastics, possible intrusive features and large-scale Tertiary channels/entry points which overlie older strike-slip zones and segment the ridge along its length. Displaying considerable complexity and structural variation along strike, seismic based interpretations provide models ranging from a shallow basement horst to preservation of significant thicknesses of Paleozoic and Mesozoic sediments. Recent work by Eni identifies the potential for prospective section to be preserved on the Erris Ridge resulting in two large prospects, Fiachra and Conn, and a number of leads being identified. The Fiachra Prospect is a large fault assisted four-way dip closure situated within a crestal location on the Erris Ridge and forms the focus of an exploration well planned for 2010. An early Triassic sandstone play (Sherwood Sandstone Group equivalent) is prognosed as the primary reservoir target interval, although several secondary targets also exist. After an intensive evaluation programme the critical geological risk remains modelling the presence of sedimentary section with reservoir potential in areas where the quality of seismic data is very poor. However, it is concluded that Licence 7/97 possesses good potential for deepwater frontier exploration; the Corrib gas field to the south of the licence area and the Dooish gas discovery to the north indicate the potential for a favourable location within an effective petroleum system.

We presented evidence that the WASP-14 exoplanetary system has misaligned orbital and stellar-rotational axes, with an angle of 33.1 plus or minus 7.4 degrees between their sky projections. At the time of this publication, WASP-14 was the third system known to have a significant spin-orbit misalignment, and all three systems had super- Jupiter planets and eccentric orbits. Therefore we hypothesized that the migration and subsequent orbital evolution of massive, eccentric exoplanets is somehow different from that of less massive close-in Jupiters, the majority of which have well-aligned orbits., United States. National Aeronautics and Space Administration (NASA Origins of Solar Systems program, award NNX09AD36G), United States. National Aeronautics and Space Administration (NASA Origins of Solar Systems program, award NNX09AB33G), National Science Foundation (U.S.) (NSF Astronomy and Astrophysics Postdoctoral Fellow, grant AST-0702821), Massachusetts Institute of Technology (Class of 1942), Netherlands Organization for Scientific Research (Rubicon fellowship), United States. National Aeronautics and Space Administration (NASA grant NNG05GK92G), University of California, Berkeley (Space Sciences Laboratory,Townes Postdoctoral Fellowship)

Traffic injuries are an important problem in low income countries. In Uganda road traffic is the largest single cause of injury in Kampala; pedestrians, and children are most affected. Pedestrian injury affects school children in Uganda.To determine the overall risk of pedestrian traffic injury among school children in Kawempe, Uganda.A cohort was assembled at 35 primary schools and followed for 3 terms. Ten of the schools had participated in previous injury programs, others were systematically selected. Injuries were recorded by teachers using a questionnaire. Data collected included ID, school, age, grade, gender, incident date, vehicle type, and injury outcome. Demographic characteristics are described and cumulative incidences calculated.The cohort included 8,165 children (49% male) from 35 primary schools. The mean age was 9 years (Sd=2.78). Of the 35 schools, 92% were day; the others mixed day and boarding. 53 children (27 girls) were involved in a traffic incident. 25% of the injuries reported were serious and warranted care in a health facility. No deaths occurred. Forty % of incidents involved commercial motorcycles, 41% bicycles, 9% cars, 8% taxis, and 2% trucks. The cumulative incidence was 0.168% each term. Over the 3 terms of the year the cumulative incidence was 0.5 +/- 0.02. There were no gender differences in the cumulative incidence.Each school year about 1/2 % of Kawempe school children are involved in a traffic incident. Interventions are necessary to reduce the unacceptably high incidents of pedestrian traffic. Interventions to alleviate this situation including safer routes, teaching skills of road crossing to children as well as better regulation and road safety education to two wheelers could reduce the unacceptably high incidents of pedestrian traffic injury.

Poly(lactide-co-glycolide) (PLGA) has been widely used for bone tissue regeneration. However, it lacks hydrophilicity, bioactivity and sufficient mechanical strength and its acidic degradation by-products can lead to pH decrease in the vicinity of the implants. Mesoporous bioactive glass (MBG) with highly ordered structure (pore size 2-50nm) possesses higher bioactivity than non-mesoporous bioactive glass (BG). The aim of this study is to investigate the effect of MBG on the mechanical strength, in vitro degradation, bioactivity, cellular response and drug release of PLGA films and optimize their physicochemical, biological and drug-delivery properties for bone tissue engineering application. The surface and inner microstructure, mechanical strength and surface hydrophilicity of MBG/PLGA and BG/PLGA films were tested. Results indicated that MBG or BG was uniformly dispersed in the PLGA films. The incorporation of MBG into PLGA films significantly improved their tensile strength, modulus and surface hydrophilicity. MBG/PLGA resulted in an enhanced mechanical strength, in vitro degradation (water absorbance, weight loss and ions release), apatite-formation ability and pH stability in simulated body fluids (SBF), compared to BG/PLGA. MBG/PLGA and BG/PLGA films enhanced human osteoblastic-like cells (HOBs) attachment, spreading and proliferation compared to PLGA. HOBs differentiation was significantly upregulated when cells were cultured on 30 MBG/PLGA for 14 days, compared to 30 BG/PLGA. MBG/PLGA enhanced the accumulative release of dexamethazone (DEX) at early stages (0-200h) compared to BG/PLGA, however, after 200h, DEX-release rates for MBG/PLGA was slower than that of BG/PLGA. The contents of MBG in PLGA films can control the amount of DEX released. Taken together, MBG/PLGA films possessed excellent physicochemical, biological and drug-release properties, indicating their potential application for bone tissue engineering by designing 3D scaffolds according to their corresponding compositions.

The prevalence of food allergy has increased dramatically in recent years. Tremendous research progress has been made in understanding the pathophysiological mechanisms of allergy and in identifying and characterizing food allergens. Peanut is a major food allergen source and Ara h 3 is a major peanut allergen. Using overlapping short peptides, several linear IgE-binding epitopes in Ara h 3 have been defined before. However, the structure of Ara h 3 of the native allergen is not clear and information on conformational epitopes is lacking. Structural characterization of allergens is required for understanding the allergenicity of food allergens and for the development of immunotherapeutic agents. Previously, we have reported the crystallization of Ara h 3 purified from raw peanut. Here we report the crystal structure of Ara h 3 at 1.73A resolution. Mapping of the previously defined linear epitopes on the crystal structure of Ara h 3 indicated that linear epitopes with more solvent exposure were those indicated by the literature to react with more patient sera. The structure of Ara h 3 may be used to assess the importance of conformational epitopes in further investigations.

The 11S globulins from plant seeds account for a number of major food allergens. Because of the interest in the structural basis underlying the allergenicity of food allergens, we sought to crystallize the main 11S seed storage protein from almond ( Prunus dulcis). Prunin-1 (Pru1) was purified from defatted almond flour by water extraction, cryoprecipitation, followed by sequential anion exchange, hydrophobic interaction, and size exclusion chromatography. Single crystals of Pru1 were obtained in a screening with a crystal screen kit, using the hanging-drop vapor diffusion method. Diffraction quality crystals were grown after optimization. The Pru1 crystals diffracted to at least 3.0 A and belong to the tetragonal space group P4(1)22, with unit cell parameters of a = b = 150.912 A, c = 165.248 A. Self-rotation functions and molecular replacement calculations showed that there are three molecules in the asymmetry unit with water content of 51.41%. The three Pru1 protomers are related by a noncrystallographic 3-fold axis and they form a doughnut-shaped trimer. Two prunin trimers form a homohexamer. Elucidation of prunin structure will allow further characterization of the allergenic features of the 11S protein allergens at the molecular level.

Detecting people around unmanned ground vehicles (UGVs) to facilitate safe operation of UGVs is one of the highest priority issues in the development of perception technology for autonomous navigation. Research to date has not achieved the detection ranges or reliability needed in deployed systems to detect upright pedestrians in flat, relatively uncluttered terrain, let alone in more complex environments and with people in postures that are more difficult to detect. Range data is essential to solve this problem. Combining range data with high resolution imagery may enable higher performance than range data alone because image appearance can complement shape information in range data and because cameras may offer higher angular resolution than typical range sensors. This makes stereo vision a promising approach for several reasons: image resolution is high and will continue to increase, the physical size and power dissipation of the cameras and computers will continue to decrease, and stereo cameras provide range data and imagery that are automatically spatially and temporally registered. We describe a stereo vision-based pedestrian detection system, focusing on recent improvements to a shape-based classifier applied to the range data, and present frame-level performance results that show great promise for the overall approach.

Background and objectives: Kidney transplantation is the most desired and cost-effective modality of renal replacement therapy for patients with irreversible chronic kidney failure (end-stage renal disease, stage 5 chronic kidney disease). Despite emerging evidence that the best outcomes accrue to patients who receive a transplant early in the course of renal replacement therapy, only 2.5% of incident patients with end-stage renal disease undergo transplantation as their initial modality of treatment, a figure largely unchanged for at least a decade. Design, setting, participants, & measurements: The National Kidney Foundation convened a Kidney Disease Outcomes Quality Initiative (KDOQI) conference in Washington, DC, March 19 through 20, 2007, to examine the issue. Fifty-two participants representing transplant centers, dialysis providers, and payers were divided into three work groups to address the impact of early transplantation on the chronic kidney disease paradigm, educational needs of patients and professionals, and finances of renal replacement therapy. Results: Participants explored the benefits of early transplantation on costs and outcomes, identified current barriers (at multiple levels) that impede access to early transplantation, and recommended specific interventions to overcome those barriers. Conclusions: With implementation of early education, referral to a transplant center coincident with creation of vascular access, timely transplant evaluation, and identification of potential living donors, early transplantation can be an option for substantially more patients with chronic kidney disease.

Ginkgo biloba, a well known ;living fossil' native to China, is grown worldwide as an ornamental shade plant. Medicinal and nutritional uses of G. biloba in Asia have a long history. However, ginkgo seed proteins have not been well studied at the biochemical and molecular level. In this study, the G. biloba 11S seed storage protein ginnacin was purified by sequential anion-exchange and gel-filtration chromatography. A crystallization screen was performed and well diffracting single crystals were obtained by the vapor-diffusion method. A molecular-replacement structural solution has been obtained. There are six protomers in an asymmetric unit. Structure refinement is currently in progress.

The crystal structure of HI0827 from Haemophilus influenzae Rd KW20, initially annotated "hypothetical protein" in sequence databases, exhibits an acyl-coenzyme A (acyl-CoA) thioesterase "hot dog" fold with a trimer of dimers oligomeric association, a novel assembly for this enzyme family. In studies described in the preceding paper [Zhuang, Z., Song, F., Zhao, H., Li, L., Cao, J., Eisenstein, E., Herzberg, O., and Dunaway-Mariano, D. (2008) Biochemistry 47, 2789-2796], HI0827 is shown to be an acyl-CoA thioesterase that acts on a wide range of acyl-CoA compounds. Two substrate binding sites are located across the dimer interface. The binding sites are occupied by two CoA molecules, one with full occupancy and the second only partially occupied. The CoA molecules, acquired from HI0827-expressing Escherichia coli cells, remained tightly bound to the enzyme through the protein purification steps. The difference in CoA occupancies indicates a different substrate affinity for each of the binding sites, which in turn implies that the enzyme might be subject to allosteric regulation. Mutagenesis studies have shown that the replacement of the putative catalytic carboxylate Asp44 with an alanine residue abolishes activity. The impact of this mutation is seen in the crystal structure of D44A HI0827. Whereas the overall fold and assembly of the mutant protein are the same as those of the wild-type enzyme, the CoA ligands are absent. The dimer interface is perturbed, and the channel that accommodates the thioester acyl chain is more open and wider than that observed in the wild-type enzyme. A model of intact substrate bound to wild-type HI0827 provides a structural rationale for the broad substrate range.