Assessment of cefazolin and cefuroxime tissue penetration by using a continuous intravenous infusion.

A continuous intravenous infusion was used to assess the tissue penetration of cefazolin (14 subjects) and cefuroxime (15 subjects) in orthopedic surgery patients. Subjects were randomly assigned to receive a continuous intravenous infusion of cefazolin (mean, 178.6 mg/h) orcefuroxime (mean, 330.0 mg/h) at a rate estimated to achieve a target steady-state total concentration of 50 micrograms/ml in serum. The infusion was initiated 12 to 14 h before surgery, and blood and muscle tissue samples were collected intraoperatively at the times of incision and wound closure. Although there was a significant difference between the free concentrations ofcefazolin (at incision, 9.3 micrograms/ml; at closure, 9.2 micrograms/ml) and cefuroxime in serum (at incision, 26.9 micrograms/ml; at closure, 31.8 micrograms/ml), there was no difference in the total concentrations in muscle at either surgical incision (cefazolin, 6.1 micrograms/g; cefuroxime, 5.6 micrograms/g) or wound closure (cefazolin, 7.7 micrograms/g; cefuroxime, 7.4 micrograms/g). There was a significant correlation between the pooled free serum and total muscle concentrations for cefazolin (P = 0.001); however, there was no correlation between these variables with the pooledcefuroxime data (P = 0.403). These findings indicate that the free drug concentration in serum alone is not consistently predictive of the total concentration of cephalosporin in muscle

Reflection Spectroscopy of the Black Hole Binary XTE J1752-223 in its Long-Stable Hard State

We present a detailed spectral analysis of the Black Hole Binary XTE J1752-223 in the hard state of its 2009 outburst. Regular monitoring of this source by RXTE provided high signal-to-noise spectra along the outburst rise and decay. During one full month this source stalled at $\sim$30\% of its peak count rate at a constant hardness and intensity. By combining all the data in this exceptionally-stable hard state, we obtained an aggregate PCA spectrum (3-45 keV) with 100 million counts, and a corresponding HEXTE spectrum (20-140 keV) with 5.8 million counts. Implementing a version of our reflection code with a physical model for Comptonization, we obtain tight constraints on important physical parameters for this system. In particular, the inner accretion disk is measured very close in, at $R_\mathrm{in}=1.7\pm0.4$ $R_g$. Assuming $R_\mathrm{in}=R_\mathrm{ISCO}$, we find a relatively high black hole spin ($a_*=0.92\pm0.06$). Imposing a lamppost geometry, we obtain a low inclination ($i=35\pm4$ deg), which agrees with the upper limit found in the radio ($i<49$ deg). However, we note that this model cannot be statistically distinguished from a non-lamppost model with free emissivity index, for which the inclination is markedly higher. Additionally, we find a relatively cool corona ($57-70$ keV), and large iron abundance ($3.3-3.7$ solar). We further find that properly accounting for Comptonization of the reflection emission improves the fit significantly and causes an otherwise low reflection fraction ($\sim 0.2-0.3$) to increase by an order of magnitude, in line with geometrical expectations for a lamppost corona. We compare these results with similar investigations reported for GX 339-4 in its bright hard state.Comment: Accepted for publication in ApJ. 11 pages, 7 figure

Combining ultraconserved elements and mtDNA data to uncover lineage diversity in a Mexican highland frog (Sarcohyla; Hylidae)

Molecular studies have uncovered significant diversity in the Mexican Highlands, leading to the description of many new endemic species. DNA approaches to this kind of species discovery have included both mitochondrial DNA (mtDNA) sequencing and multilocus genomic methods. While these marker types have often been pitted against one another, there are benefits to deploying them together, as linked mtDNA data can provide the bridge between uncovering lineages through rigorous multilocus genomic analysis and identifying lineages through comparison to existing mtDNA databases. Here, we apply one class of multilocus genomic marker, ultraconserved elements (UCEs), and linked mtDNA data to a species complex of frogs (Sarcohyla bistincta, Hylidae) found in the Mexican Highlands. We generated data from 1,891 UCEs, which contained 1,742 informative SNPs for S. bistincta and closely related species and captured mitochondrial genomes for most samples. Genetic analyses based on both whole loci and SNPs agree there are six to seven distinct lineages within what is currently described as S. bistincta. Phylogenies from UCEs and mtDNA mostly agreed in their topologies, and the few differences suggested a more complex evolutionary history of the mtDNA marker. Our study demonstrates that the Mexican Highlands still hold substantial undescribed diversity, making their conservation a particularly urgent goal. The Trans-Mexican Volcanic Range stands out as a significant geographic feature in Sarcohyla and may have acted as a dispersal corridor for S. bistincta to spread to the north. Combining multilocus genomic data with linked mtDNA data is a useful approach for identifying potential new species and associating them with already described taxa, which will be especially important in groups with undescribed subadult phenotypes and cryptic species

Structure and Recognition of a Novel HIV-1 gp120-gp41 Interface Antibody that Caused MPER Exposure through Viral Escape

A comprehensive understanding of the regions on HIV-1 envelope trimers targeted by broadly neutralizing antibodies may contribute to rational design of an HIV-1 vaccine. We previously identified a participant in the CAPRISA cohort, CAP248, who developed trimer-specific antibodies capable of neutralizing 60% of heterologous viruses at three years post-infection. Here, we report the isolation by B cell culture of monoclonal antibody CAP248-2B, which targets a novel membrane proximal epitope including elements of gp120 and gp41. Despite low maximum inhibition plateaus, often below 50% inhibitory concentrations, the breadth of CAP248-2B significantly correlated with donor plasma. Site-directed mutagenesis, X-ray crystallography, and negative-stain electron microscopy 3D reconstructions revealed how CAP248-2B recognizes a cleavage-dependent epitope that includes the gp120 C terminus. While this epitope is distinct, it overlapped in parts of gp41 with the epitopes of broadly neutralizing antibodies PGT151, VRC34, 35O22, 3BC315, and 10E8. CAP248-2B has a conformationally variable paratope with an unusually long 19 amino acid light chain third complementarity determining region. Two phenylalanines at the loop apex were predicted by docking and mutagenesis data to interact with the viral membrane. Neutralization by CAP248-2B is not dependent on any single glycan proximal to its epitope, and low neutralization plateaus could not be completely explained by N- or O-linked glycosylation pathway inhibitors, furin co-transfection, or pre-incubation with soluble CD4. Viral escape from CAP248-2B involved a cluster of rare mutations in the gp120-gp41 cleavage sites. Simultaneous introduction of these mutations into heterologous viruses abrogated neutralization by CAP248-2B, but enhanced neutralization sensitivity to 35O22, 4E10, and 10E8 by 10-100-fold. Altogether, this study expands the region of the HIV-1 gp120-gp41 quaternary interface that is a target for broadly neutralizing antibodies and identifies a set of mutations in the gp120 C terminus that exposes the membrane-proximal external region of gp41, with potential utility in HIV vaccine design

Developmental Pathway of the MPER-Directed HIV-1-Neutralizing Antibody 10E8

Antibody 10E8 targets the membrane-proximal external region (MPER) of HIV-1 gp41, neutralizes >97% of HIV-1 isolates, and lacks the auto-reactivity often associated with MPER-directed antibodies. The developmental pathway of 10E8 might therefore serve as a promising template for vaccine design, but samples from time-of-infection—often used to infer the B cell record—are unavailable. In this study, we used crystallography, next-generation sequencing (NGS), and functional assessments to infer the 10E8 developmental pathway from a single time point. Mutational analysis indicated somatic hypermutation of the 2nd-heavy chain-complementarity determining region (CDR H2) to be critical for neutralization, and structures of 10E8 variants with V-gene regions reverted to genomic origin for heavy-and-light chains or heavy chain-only showed structural differences >2 Å relative to mature 10E8 in the CDR H2 and H3. To understand these developmental changes, we used bioinformatic sieving, maximum likelihood, and parsimony analyses of immunoglobulin transcripts to identify 10E8-lineage members, to infer the 10E8-unmutated common ancestor (UCA), and to calculate 10E8-developmental intermediates. We were assisted in this analysis by the preservation of a critical D-gene segment, which was unmutated in most 10E8-lineage sequences. UCA and early intermediates weakly bound a 26-residue-MPER peptide, whereas HIV-1 neutralization and epitope recognition in liposomes were only observed with late intermediates. Antibody 10E8 thus develops from a UCA with weak MPER affinity and substantial differences in CDR H2 and H3 from the mature 10E8; only after extensive somatic hypermutation do 10E8-lineage members gain recognition in the context of membrane and HIV-1 neutralization

Reflection Spectroscopy of the Black Hole Binary XTE J1752−223 in Its Long-stable Hard State

We present a detailed spectral analysis of the black hole binary XTE J1752−223 in the hard state of its 2009 outburst. Regular monitoring of this source by the Rossi X-ray Timing Explorer mission provided high signal-to-noise spectra along the outburst rise and decay. During one full month this source stalled at ~30% of its peak count rate at a constant hardness and intensity. By combining all the data in this exceptionally stable hard state, we obtained an aggregate proportional counter array spectrum (3–45 keV) with 100 million counts, and a corresponding high energy X-ray timing experiment spectrum (20–140 keV) with 5.8 million counts. Implementing a version of our reflection code with a physical model for Comptonization, we obtain tight constraints on important physical parameters for this system. In particular, the inner accretion disk is measured very close in, at R_(in) = 1.7 ± 0.4 R_g . Assuming R_(in_ = R_(ISCO), we find a relatively high black hole spin (a_* = 0.92 ± 0.06). Imposing a lamppost geometry, we obtain a low inclination (i = 35° ± 4°), which agrees with the upper limit found in the radio (i < 49°). However, we note that this model cannot be statistically distinguished from a non-lamppost model with a free emissivity index, for which the inclination is markedly higher. Additionally, we find a relatively cool corona (57–70 keV) and large iron abundance (3.3–3.7 solar). We further find that properly accounting for Comptonization of the reflection emission improves the fit significantly and causes an otherwise low reflection fraction (~0.2–0.3) to increase by an order of magnitude, in line with geometrical expectations for a lamppost corona. We compare these results with similar investigations reported for GX 339−4 in its bright hard state

Double Spin Asymmetry of Electrons from Heavy Flavor Decays in p+p Collisions at sqrt(s)=200 GeV

We report on the first measurement of double-spin asymmetry, A_LL, of electrons from the decays of hadrons containing heavy flavor in longitudinally polarized p+p collisions at sqrt(s)=200 GeV for p_T= 0.5 to 3.0 GeV/c. The asymmetry was measured at mid-rapidity (|eta|<0.35) with the PHENIX detector at the Relativistic Heavy Ion Collider. The measured asymmetries are consistent with zero within the statistical errors. We obtained a constraint for the polarized gluon distribution in the proton of |Delta g/g(log{_10}x= -1.6^+0.5_-0.4, {mu}=m_T^c)|^2 < 0.033 (1 sigma), based on a leading-order perturbative-quantum-chromodynamics model, using the measured asymmetry.Comment: 385 authors, 17 pages, 15 figures, 5 tables. Submitted to Phys. Rev. D. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm