Nuclear targeting of the serine protease granzyme A (fragmentin-1)

Cytolytic granule-mediated target cell killing is effected in part through synergistic action of the membrane-acting protein perforin and serine proteases such as granzymes A (GrA) or B (GrB). In the present study we examine GrA cellular entry and nuclear uptake in intact mouse myeloid FDC-P1 cells exposed to perforin using confocal laser scanning microscopy, as well as reconstitute GrA nuclear uptake in vitro. GrA alone was found to be able to enter the cytoplasm of intact cells but did not accumulate in nuclei. In the presence of perforin, it specifically accumulated in the cell nuclei, with maximal levels about 2.5 times those in the cytoplasm after 2. 5 hours. In vitro, GrA accumulated in the nucleus and nucleolus maximally to levels that were four- and sixfold, respectively, those in the cytoplasm. In contrast, the active form of the apoptotic cysteine protease CPP32 did not accumulate in nuclei in vitro. Nuclear/nucleolar import of GrA in vitro was independent of ATP and not inhibitable by the non-hydrolyzable GTP analog GTPgammaS, but was dependent on exogenously added cytosol. Importantly, GrA was found to be able to accumulate in the nucleus of semi-intact cells in the presence of the nuclear envelope-permeabilizing detergent CHAPS, implying that the mechanism of nuclear accumulation was through binding to insoluble factors in the nucleus. GrB was found for the first time to be similar in this regard. The results support the contention that GrA and GrB accumulate in the nucleus through a novel nuclear import pathway, and that this is integral to induction of the nuclear changes associated with cytolytic granule-mediated apoptosis.David A. Jans, Lyndall J. Briggs, Patricia Jans, Christopher J. Froelich, Gayathri Parasivam, Sharad Kumar, Vivien R. Sutton and Joseph A. Trapan

The influence of taphonomy on histological and isotopic analyses of treated and untreated buried modern human bone

The chemical (e.g., preservation/embalming) treatment of skeletal remains can reduce overall DNA quality and quantity. The histological and stable isotope examination of treated and untreated human remains improves our understanding of how chemical preservatives impact bone diagenesis and will determine if chemical treatment adversely affects stable isotope ratio analysis of collagen. Fidelity in the application(s) of stable isotope interpretations requires that the isotope delta (δ) values have not been altered postmortem. Re-associated antimeres and refits of chemically treated and untreated rib and long bones from eight casualties [thin-sectioned human bone (n = 43) and collagen extraction/stable isotope analysis (n = 42)] from the World War II Battle of Tarawa were examined to compare skeletal elements from the same individual that had different taphonomic histories. Histological analyses included scoring upon the Oxford Histological Index (OHI) and Birefringence scale, recording microbial invasion, and general observations. The collected data were analyzed via simple descriptive statistics and paired samples t-tests. Treated remains scored higher on the OHI and for Birefringence, indicating that bone quality was good to excellent. The untreated samples scored lower on the OHI and Birefringence scales suggesting poorer preservation than the treated remains. Histology results were supported by the isotope sample preparation results: the collagen % yield was higher for treated bone than untreated bone. Additionally, chemical preservation had no meaningful impact on isotope δ values of treated and untreated remains from the same element or pair-matched elements. Overall, treated remains exhibited good preservation while untreated remains exhibit poorer preservation with significant microfocal destruction to the extent that little histological analyses can be applied. Stable isotope ratio analysis is viable for both treated and untreated remains indicating this testing modality likely can be used for most treated remains, regardless of origin

Investigation of the Exclusive 3He(e,e'pp)n Reaction

Cross sections for the 3He(e,e'pp)n reaction were measured over a wide range of energy and three- momentum transfer. At a momentum transfer q=375 MeV/c, data were taken at transferred energies omega ranging from 170 to 290 MeV. At omega=220 MeV, measurements were performed at three q values (305, 375, and 445 MeV/c). The results are presented as a function of the neutron momentum in the final-state, as a function of the energy and momentum transfer, and as a function of the relative momentum of the two-proton system. The data at neutron momenta below 100 MeV/c, obtained for two values of the momentum transfer at omega=220 MeV, are well described by the results of continuum-Faddeev calculations. These calculations indicate that the cross section in this domain is dominated by direct two-proton emission induced by a one-body hadronic current. Cross section distributions determined as a function of the relative momentum of the two protons are fairly well reproduced by continuum-Faddeev calculations based on various realistic nucleon-nucleon potential models. At higher neutron momentum and at higher energy transfer, deviations between data and calculations are observed that may be due to contributions of isobar currents.Comment: 14 pages, 1 table, 17 figure

Radiation damage in the LHCb vertex locator

The LHCb Vertex Locator (VELO) is a silicon strip detector designed to reconstruct charged particle trajectories and vertices produced at the LHCb interaction region. During the first two years of data collection, the 84 VELO sensors have been exposed to a range of fluences up to a maximum value of approximately 45 × 1012 1 MeV neutron equivalent (1 MeV neq). At the operational sensor temperature of approximately −7 °C, the average rate of sensor current increase is 18 μA per fb−1, in excellent agreement with predictions. The silicon effective bandgap has been determined using current versus temperature scan data after irradiation, with an average value of Eg = 1.16±0.03±0.04 eV obtained. The first observation of n+-on-n sensor type inversion at the LHC has been made, occurring at a fluence of around 15 × 1012 of 1 MeV neq. The only n+-on-p sensors in use at the LHC have also been studied. With an initial fluence of approximately 3 × 1012 1 MeV neq, a decrease in the Effective Depletion Voltage (EDV) of around 25 V is observed. Following this initial decrease, the EDV increases at a comparable rate to the type inverted n+-on-n type sensors, with rates of (1.43±0.16) × 10−12 V/ 1 MeV neq and (1.35±0.25) × 10−12 V/ 1 MeV neq measured for n+-on-p and n+-on-n type sensors, respectively. A reduction in the charge collection efficiency due to an unexpected effect involving the second metal layer readout lines is observed

ICMSF Methods Studies. XV. Comparison of Four Media and Methods for Enumerating Staphylococcus aureus in Powdered Milk.

Four media were examined for their usefulness in enumerating Staphylococcus aureus inoculated (a) into milk that was then dried or (b) directly into dried milk powder. In all, seven strains of S. aureus were inoculated individually into each preparation and were enumerated after two periods of storage (18 to 19 d and 60 to 61 d). Fourteen laboratories from twelve countries participated in the comparison which found that direct plating on agar medium in 14-cm petri dishes may be as useful as enrichment followed by streaking. Plating on Baird-Parker medium or on Hauschild pork plasma fibrinogen medium and a MPN method using Giolitti and Cantoni's broth with Tween 80 were equally sensitive for enumerating S. aureus in dried milk powder. The use of Hauschild medium may eliminate the need for supplementary tests to confirm colonies as S. aureus , but in some cases was found to fail in some laboratories. Giolitti and Cantoni's broth without Tween 80 generally was less useful than the three other media for enumerating S. aureus . S. aureus inoculated into milk that was then dried survived longer than when inoculated into dried milk

A computational analysis of lower bounds for big bucket production planning problems

In this paper, we analyze a variety of approaches to obtain lower bounds for multi-level production planning problems with big bucket capacities, i.e., problems in which multiple items compete for the same resources. We give an extensive survey of both known and new methods, and also establish relationships between some of these methods that, to our knowledge, have not been presented before. As will be highlighted, understanding the substructures of difficult problems provide crucial insights on why these problems are hard to solve, and this is addressed by a thorough analysis in the paper. We conclude with computational results on a variety of widely used test sets, and a discussion of future research

First measurements of the ^16O(e,e'pn)^14N reaction

This paper reports on the first measurement of the ^16O(e,e'pn)^14N reaction. Data were measured in kinematics centred on a super-parallel geometry at energy and momentum transfers of 215 MeV and 316 MeV/c. The experimental resolution was sufficient to distinguish groups of states in the residual nucleus but not good enough to separate individual states. The data show a strong dependence on missing momentum and this dependence appears to be different for two groups of states in the residual nucleus. Theoretical calculations of the reaction using the Pavia code do not reproduce the shape or the magnitude of the data.Comment: 10 pages, 11 figures, 2 tables, Accepted for publication in EPJ

Measurement of the 12C(e,e'p)11B Two-Body Breakup Reaction at High Missing Momentum Values

The five-fold differential cross section for the 12C(e,e'p)11B reaction was determined over a missing momentum range of 200-400 MeV/c, in a kinematics regime with Bjorken x > 1 and Q2 = 2.0 (GeV/c)2. A comparison of the results and theoretical models and previous lower missing momentum data is shown. The theoretical calculations agree well with the data up to a missing momentum value of 325 MeV/c and then diverge for larger missing momenta. The extracted distorted momentum distribution is shown to be consistent with previous data and extends the range of available data up to 400 MeV/c.Comment: 12 pages, 1 table and 3 figures for submission to Journal Physics

High-precision Studies of the 3^{\bf{3}}He(e,e′^{\bf{\prime}}p) Reaction at the Quasielastic Peak

Precision studies of the reaction $^{3}$He(e,e$^\prime$p) using the three-spectrometer facility at the Mainz microtron MAMI are presented. All data are for quasielastic kinematics at $|\vec{q} | =685$ MeV/c. Absolute cross sections were measured at three electron kinematics. For the measured missing momenta range from 10 to 165 MeV/c, no strength is observed for missing energies higher than 20 MeV. Distorted momentum distributions were extracted for the two-body breakup and the continuum. The longitudinal and transverse behavior was studied by measuring the cross section for three photon polarizations. The longitudinal and transverse nature of the cross sections is well described by a currently accepted and widely used prescription of the off-shell electron-nucleon cross-section. The results are compared to modern three-body calculations and to previous data.Comment: 4 pages, 3 figures. Submitted for publication in Phys. Rev. Let

Performance of the LHCb vertex locator

The Vertex Locator (VELO) is a silicon microstrip detector that surrounds the proton-proton interaction region in the LHCb experiment. The performance of the detector during the first years of its physics operation is reviewed. The system is operated in vacuum, uses a bi-phase CO2 cooling system, and the sensors are moved to 7 mm from the LHC beam for physics data taking. The performance and stability of these characteristic features of the detector are described, and details of the material budget are given. The calibration of the timing and the data processing algorithms that are implemented in FPGAs are described. The system performance is fully characterised. The sensors have a signal to noise ratio of approximately 20 and a best hit resolution of 4 μm is achieved at the optimal track angle. The typical detector occupancy for minimum bias events in standard operating conditions in 2011 is around 0.5%, and the detector has less than 1% of faulty strips. The proximity of the detector to the beam means that the inner regions of the n+-on-n sensors have undergone space-charge sign inversion due to radiation damage. The VELO performance parameters that drive the experiment's physics sensitivity are also given. The track finding efficiency of the VELO is typically above 98% and the modules have been aligned to a precision of 1 μm for translations in the plane transverse to the beam. A primary vertex resolution of 13 μm in the transverse plane and 71 μm along the beam axis is achieved for vertices with 25 tracks. An impact parameter resolution of less than 35 μm is achieved for particles with transverse momentum greater than 1 GeV/c