Truncated Enterohemorrhagic \u3ci\u3eEscherichia coli\u3c/i\u3e (EHEC) O157:H7 Intimin (EaeA) Fusion Proteins Promote Adherence of EHEC Strains to HEp-2 Cells

Intimin, the product of the eaeA gene in enterohemorrhagic Escherichia coli O157:H7 (EHEC), is required for intimate adherence of these organisms to tissue culture cells and formation of the attaching and effacing lesion in the gnotobiotic pig. Because of the importance of intimin in the pathogenesis of EHEC O157:H7 infection in this animal model, we began a structure-function analysis of EaeA. For this purpose, we constructed amino-terminal fusions of the intimin protein with six histidine residues to form two independent fusions. The longer fusion, RIHisEae, contained 900 of the 935 predicted amino acids and included all but the extreme amino terminus. The second fusion, RVHdHisEae, consisted of the carboxyl two-thirds of the protein. Purified extracts of either construct enhanced binding of wild-type 86-24 to HEp-2 cells and conferred HEp-2 cell adherence on 86-24eaeΔ10, an eaeA deletion mutant, and B2F1, an EHEC O91:H21 eaeA mutant strain. When 86-24eaeΔ10 was transformed with either of the plasmids encoding the intimin fusion proteins, the transformant behaved like the wild-type parent strain and displayed localized adherence to HEp-2 cells, with positive fluorescent-actin staining. In addition, polyclonal antisera raised against RIHisEae reacted with both fusion constructs and recognized an outer membrane protein of the same mass as intimin (97 kDa) in EHEC and enteropathogenic E. coli but not E. coli K-12. The intimin-specific antisera also blocked adherence of EHEC to HEp-2 cells. Thus, intimin (i) is a 97-kDa outer membrane protein in EHEC that serves as a requisite adhesin for attachment of the bacteria to epithelial cells, even when the protein is truncated by one-third at its amino terminus and (ii) can be added exogenously to specifically facilitate HEp-2 cell adherence of EHEC but not E. coli K-12

Truncated Enterohemorrhagic \u3ci\u3eEscherichia coli\u3c/i\u3e (EHEC) O157:H7 Intimin (EaeA) Fusion Proteins Promote Adherence of EHEC Strains to HEp-2 Cells

Intimin, the product of the eaeA gene in enterohemorrhagic Escherichia coli O157:H7 (EHEC), is required for intimate adherence of these organisms to tissue culture cells and formation of the attaching and effacing lesion in the gnotobiotic pig. Because of the importance of intimin in the pathogenesis of EHEC O157:H7 infection in this animal model, we began a structure-function analysis of EaeA. For this purpose, we constructed amino-terminal fusions of the intimin protein with six histidine residues to form two independent fusions. The longer fusion, RIHisEae, contained 900 of the 935 predicted amino acids and included all but the extreme amino terminus. The second fusion, RVHdHisEae, consisted of the carboxyl two-thirds of the protein. Purified extracts of either construct enhanced binding of wild-type 86-24 to HEp-2 cells and conferred HEp-2 cell adherence on 86-24eaeΔ10, an eaeA deletion mutant, and B2F1, an EHEC O91:H21 eaeA mutant strain. When 86-24eaeΔ10 was transformed with either of the plasmids encoding the intimin fusion proteins, the transformant behaved like the wild-type parent strain and displayed localized adherence to HEp-2 cells, with positive fluorescent-actin staining. In addition, polyclonal antisera raised against RIHisEae reacted with both fusion constructs and recognized an outer membrane protein of the same mass as intimin (97 kDa) in EHEC and enteropathogenic E. coli but not E. coli K-12. The intimin-specific antisera also blocked adherence of EHEC to HEp-2 cells. Thus, intimin (i) is a 97-kDa outer membrane protein in EHEC that serves as a requisite adhesin for attachment of the bacteria to epithelial cells, even when the protein is truncated by one-third at its amino terminus and (ii) can be added exogenously to specifically facilitate HEp-2 cell adherence of EHEC but not E. coli K-12

Fibronectin as a Prognostic Indicator in Portal Hypertension

Plasma fibronectin levels were measured in 33 patients with portal hypertension and c6mpared with modified Child’s grading and a previously described prognostic index. Outcome at one year from blood sampling was recorded. Mean plasma fibronectin level was 304.1 mg/ml (sem 24.3) and significantly lower levels were found in patients who had had a variceal bleed within the previous seven days. Plasma fibronectin levels tended to be lower in patients with poor liver function as assessed by modified Child’s grading but this did not achieve statistical significance. Plasma fibronectin alone was not an accurate predictor of one year survival in these patients but only one of seven patients who had a plasma fibronectin level below 300mg/l in association with a poor prognostic index survived for one year

Quantum Information Processing with Ferroelectrically Coupled Quantum Dots

I describe a proposal to construct a quantum information processor using ferroelectrically coupled Ge/Si quantum dots. The spin of single electrons form the fundamental qubits. Small (<10 nm diameter) Ge quantum dots are optically excited to create spin polarized electrons in Si. The static polarization of an epitaxial ferroelectric thin film confines electrons laterally in the semiconductor; spin interactions between nearest neighbor electrons are mediated by the nonlinear process of optical rectification. Single qubit operations are achieved through "g-factor engineering" in the Ge/Si structures; spin-spin interactions occur through Heisenberg exchange, controlled by ferroelectric gates. A method for reading out the final state, while required for quantum computing, is not described; electronic approaches involving single electron transistors may prove fruitful in satisfying this requirement.Comment: 10 pages, 3 figure

Using computer graphics to design Space Station Freedom viewing

Viewing requirements were identified early in the Space Station Freedom program for both direct viewing via windows and indirect viewing via cameras and closed-circuit television (CCTV). These requirements reside in NASA Program Definition and Requirements Document (PDRD), Section 3: Space Station Systems Requirements. Currently, analyses are addressing the feasibility of direct and indirect viewing. The goal of these analyses is to determine the optimum locations for the windows, cameras, and CCTV's in order to meet established requirements, to adequately support space station assembly, and to operate on-board equipment. PLAID, a three-dimensional computer graphics program developed at NASA JSC, was selected for use as the major tool in these analyses. PLAID provides the capability to simulate the assembly of the station as well as to examine operations as the station evolves. This program has been used successfully as a tool to analyze general viewing conditions for many Space Shuttle elements and can be used for virtually all Space Station components. Additionally, PLAID provides the ability to integrate an anthropometric scale-modeled human (representing a crew member) with interior and exterior architecture

Durable response to palbociclib and letrozole in ovarian cancer with CDKN2A loss.

Alterations of the Retinoblastoma (Rb) pathway are frequent in ovarian cancer, typically resulting from CDKN2A down-regulation, CCNE1 amplification, CCND1/2 amplification, and RB1 loss. However, bi-allelic CDKN2A mutation or homozygous deletion is a very rare event, concerning less than 5% of patients.Initial trials with palbociclib in serous ovarian cancer have shown very modest benefit in unselected patient populations, thus underlining the need for a biomarker predicting response. We report the case of a heavily pre-treated patient with a serous ovarian tumor harboring a homozygous deletion of the CDKN2A gene that derived significant, prolonged clinical benefit from palbociclib, a CDK4/6 oral inhibitor, with letrozole. Treatment with palbociclib and letrozole started on February 2018, with an ongoing response after 12 months.In conclusion, homozygous CDKN2A deletion is rare and could be used to predict response to CDK4/6 inhibitors in association with other genomic features. We encourage further trials in this direction

An infrared-submillimeter study of star-forming regions selected by the ISOSS 170um survey

Using the ISOPHOT Serendipity Survey (ISOSS) at 170um a sample of galactic star-forming regions exhibiting very cold dust temperatures (< 20 K) and high masses (> 100 M_sun) has been established. We characterise the star-forming content of five regions that were selected as potential sites for early stage high-mass star formation using SCUBA (JCMT) and Spitzer observations. In every region we identify one to four submillimeter clumps with projected sizes between 0.1 and 0.4 pc. The dust temperatures range from 11.6 to 21.3 K and the estimated clump masses are 2 to 166 M_sun. Towards the majority of submillimeter peaks we find point sources in the near- to mid-infrared. Most are interpreted as low-mass young stellar objects but we also detect very red sources. They probably represent very young and deeply embedded protostars that continue to accrete clump material and may reach higher masses. Several candidate intermediate-mass proto- or pre-main-sequence stars embedded in the clumps are identified. A subset of four clumps may be massive enough (> 100 M_sun) to form high-mass stars and accompanying clusters. The absence of stellar precursors with current masses in the high-mass regime leave the type of star formation occuring in the clumps unsettled. We confirm the presence of large fractions of cold material as derived from large-scale far-infrared measurements which dominates the emission of most clumps and suggests that the star-forming process will continue.Comment: 11 pages, 11 figures, accepted for publication in A&

Mid-infrared interferometry of the massive young stellar object NGC3603 - IRS 9A

We present observations and models for one of these MYSO candidates, NGC3603 IRS 9A. Our goal is to investigate with infrared interferometry the structure of IRS 9A on scales as small as 200AU, exploiting the fact that a cluster of O and B stars has blown away much of the obscuring foreground dust and gas. Observations in the N-band were carried out with the MIDI beam combiner attached to the VLTI. Additional interferometric observations which probe the structure of IRS 9A on larger scales were performed with an aperture mask installed in the T-ReCS instrument of Gemini South. The spectral energy distribution (SED) is constrained by the MIDI N-band spectrum and by data from the Spitzer Space Telescope. Our efforts to model the structure and SED of IRS 9A range from simple geometrical models of the brightness distribution to one- and two-dimensional radiative transfer computations. The target is resolved by T-ReCS, with an equivalent (elliptical) Gaussian width of 330mas by 280mas (2300 AU by 2000 AU). Despite this fact, a warm compact unresolved component was detected by MIDI which is possibly associated with the inner regions of a flattened dust distribution. Based on our interferometric data, no sign of multiplicity was found on scales between about 200AU and 700AU projected separation. A geometric model consisting of a warm (1000 K) ring (400 AU diameter) and a cool (140 K) large envelope provides a good fit to the data. No single model fitting all visibility and photometric data could be found, with disk models performing better than spherical models. While the data are clearly inconsistent with a spherical dust distribution they are insufficient to prove the existence of a disk but rather hint at a more complex dust distribution.Comment: 8 pages, 11 figures. Accepted for publication in A&

Faraday Rotation as a diagnostic of Galactic foreground contamination of CMB maps

The contribution from the residuals of the foreground can have a significant impact on the temperature maps of the Cosmic Microwave Background (CMB). Mostly, the focus has been on the galactic plane, when foreground cleaning has taken place. However, in this paper, we will investigate the possible foreground contamination, from sources outside the galactic plane in the CMB maps. We will analyze the correlation between the Faraday rotation map and the CMB temperature map. The Faraday rotation map is dependent on the galactic magnetic field, as well as the thermal electron density, and both may contribute to the CMB temperature. We find that the standard deviation for the mean cross correlation deviate from that of simulations at the 99.9% level. Additionally, a comparison between the CMB temperature extrema and the extremum points of the Faraday rotation is also performed, showing a general overlap between the two. Also we find that the CMB Cold Spot is located at an area of strong negative cross correlation, meaning that it may be explained by a galactic origin. Further, we investigate nearby supernova remnants in the galaxy, traced by the galactic radio loops. These super nova remnants are located at high and low galactic latitude, and thus well outside the galactic plane. We find some correlation between the Faraday Rotation and the CMB temperature, at select radio loops. This indicate, that the galactic foregrounds may affect the CMB, at high galactic latitudesComment: 13 pages, 22 figures, 6 table