Depletion induced isotropic-isotropic phase separation in suspensions of rod-like colloids

When non-adsorbing polymers are added to an isotropic suspension of rod-like colloids, the colloids effectively attract each other via depletion forces. We performed Monte Carlo simulations to study the phase diagram of such rod-polymer mixture. The colloidal rods were modelled as hard spherocylinders; the polymers were described as spheres of the same diameter as the rods. The polymers may overlap with no energy cost, while overlap of polymers and rods is forbidden. Large amounts of depletant cause phase separation of the mixture. We estimated the phase boundaries of isotropic-isotropic coexistence both, in the bulk and in confinement. To determine the phase boundaries we applied the grand canonical ensemble using successive umbrella sampling [J. Chem. Phys. 120, 10925 (2004)], and we performed a finite-size scaling analysis to estimate the location of the critical point. The results are compared with predictions of the free volume theory developed by Lekkerkerker and Stroobants [Nuovo Cimento D 16, 949 (1994)]. We also give estimates for the interfacial tension between the coexisting isotropic phases and analyse its power-law behaviour on approach of the critical point

Osmotic compression of droplets of hard rods: A computer simulation study

By means of computer simulations we study how droplets of hard, rod-like particles optimize their shape and internal structure under the influence of the osmotic compression caused by the presence of spherical particles that act as depletion agents. At sufficiently high osmotic pressures the rods that make up the drops spontaneously align to turn them into uniaxial nematic liquid crystalline droplets. The nematic droplets or "tactoids" that are formed this way are not spherical but elongated, resulting from the competition between the anisotropic surface tension and the elastic deformation of the director field. In agreement with recent theoretical predictions we find that sufficiently small tactoids have a uniform director field, whilst large ones are characterized by a bipolar director field. From the shape and director-field transformation of the droplets we are able to estimate the surface anchoring strength and an average of the elastic constants of the hard-rod nematic

Comparability of Pulmonary Nodule Size Measurements among Different Scanners and Protocols: Should Diameter Be Favorized over Volume?

BACKGROUND: To assess the impact of the lung cancer screening protocol recommended by the European Society of Thoracic Imaging (ESTI) on nodule diameter, volume, and density throughout different computed tomography (CT) scanners. METHODS: An anthropomorphic chest phantom containing fourteen different-sized (range 3-12 mm) and CT-attenuated (100 HU, -630 HU and -800 HU, termed as solid, GG1 and GG2) pulmonary nodules was imaged on five CT scanners with institute-specific standard protocols (P$_{S}$) and the lung cancer screening protocol recommended by ESTI (ESTI protocol, P$_{E}$). Images were reconstructed with filtered back projection (FBP) and iterative reconstruction (REC). Image noise, nodule density and size (diameter/volume) were measured. Absolute percentage errors (APEs) of measurements were calculated. RESULTS: Using P$_{E}$, dosage variance between different scanners tended to decrease compared to P$_{S}$, and the mean differences were statistically insignificant (p = 0.48). P$_{S}$ and P$_{E(REC)}$ showed significantly less image noise than P$_{E(FBP)}$ (p < 0.001). The smallest size measurement errors were noted with volumetric measurements in P$_{E(REC)}$ and highest with diametric measurements in P$_{E(FBP)}$. Volume performed better than diameter measurements in solid and GG1 nodules (p < 0.001). However, in GG2 nodules, this could not be observed (p = 0.20). Regarding nodule density, REC values were more consistent throughout different scanners and protocols. CONCLUSION: Considering radiation dose, image noise, nodule size, and density measurements, we fully endorse the ESTI screening protocol including the use of REC. For size measurements, volume should be preferred over diameter

Pole-to-Pole Connections : Similarities between Arctic and Antarctic Microbiomes and Their Vulnerability to Environmental Change

Acknowledgments JK acknowledges the Carl Zeiss foundation for PhD funding, the Marie-Curie COFUND-BEIPD PostDoc fellowship for PostDoc funding, FNRS travel funding and the logistical and financial support by UNIS. JK and FK acknowledge the Natural Environment Research Council (NERC) Antarctic Funding Initiative AFI-CGS-70 (collaborative gearing scheme) and logistic support from the British Antarctic Survey (BAS) for field work in Antarctica. JK and CZ acknowledge the Excellence Initiative at the University of Tübingen funded by the German Federal Ministry of Education and Research and the German Research Foundation (DFG). FH, AV, and PB received funding from MetaHIT (HEALTH-F4-2007-201052), Microbios (ERC-AdG-502 669830) and the European Molecular Biology Laboratory (EMBL). We thank members of the Bork group at EMBL for helpful discussions. We acknowledge the EMBL Genomics Core Facility for sequencing support and Y. P. Yuan and the EMBL Information Technology Core Facility for support with high-performance computing and EMBL for financial support. PC is supported by NERC core funding to the BAS “Biodiversity, Evolution and Adaptation” Team. MB was funded by Helge Ax:son Johnsons Stiftelse and PUT1317. DRD acknowledges the DFG funded project DI698/18-1 Dietrich and the Marie Curie International Research Staff Exchange Scheme Fellowship (PIRSES-GA-2011-295223). Operations in the Canadian High Arctic were supported by the Natural Sciences and Engineering Research Council of Canada (NSERC), ArcticNet and the Polar Continental Shelf Program (PCSP). We are also grateful to the TOTAL Foundation (Paris) and the UK NERC (WP 4.3 of Oceans 2025 core funding to FCK at the Scottish Association for Marine Science) for funding the expedition to Baffin Island and within this context Olivier Dargent and Dr. Pieter van West for sample collection, and the Spanish Ministry of Science and Technology through project LIMNOPOLAR (POL200606635 and CGL2005-06549-C02-01/ANT to AQ as well as CGL2005-06549-C02-02/ANT to AC, the last of these co-financed by European FEDER funds). We are grateful for funding from the MASTS pooling initiative (The Marine Alliance for Science and Technology for Scotland), funded by the Scottish Funding Council (HR09011) and contributing institutions. Supplementary Material The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fevo.2017.00137/full#supplementary-materialPeer reviewedPublisher PD

A multimodal imaging workflow for monitoring CAR T cell therapy against solid tumor from whole-body to single-cell level

CAR T cell research in solid tumors often lacks spatiotemporal information and therefore, there is a need for a molecular tomography to facilitate high-throughput preclinical monitoring of CAR T cells. Furthermore, a gap exists between macro- and microlevel imaging data to better assess intratumor infiltration of therapeutic cells. We addressed this challenge by combining 3D µComputer tomography bioluminescence tomography (µCT/BLT), light-sheet fluorescence microscopy (LSFM) and cyclic immunofluorescence (IF) staining. Methods: NSG mice with subcutaneous AsPC1 xenograft tumors were treated with EGFR CAR T cell (± IL-2) or control BDCA-2 CAR T cell (± IL-2) (n = 7 each). Therapeutic T cells were genetically modified to co-express the CAR of interest and the luciferase CBR2opt. IL-2 was administered s.c. under the xenograft tumor on days 1, 3, 5 and 7 post-therapy-initiation at a dose of 25,000 IU/mouse. CAR T cell distribution was measured in 2D BLI and 3D µCT/BLT every 3-4 days. On day 6, 4 tumors were excised for cyclic IF where tumor sections were stained with a panel of 25 antibodies. On day 6 and 13, 8 tumors were excised from rhodamine lectin-preinjected mice, permeabilized, stained for CD3 and imaged by LSFM. Results: 3D µCT/BLT revealed that CAR T cells pharmacokinetics is affected by antigen recognition, where CAR T cell tumor accumulation based on target-dependent infiltration was significantly increased in comparison to target-independent infiltration, and spleen accumulation was delayed. LSFM supported these findings and revealed higher T cell accumulation in target-positive groups at day 6, which also infiltrated the tumor deeper. Interestingly, LSFM showed that most CAR T cells accumulate at the tumor periphery and around vessels. Surprisingly, LSFM and cyclic IF revealed that local IL-2 application resulted in early-phase increased proliferation, but long-term overstimulation of CAR T cells, which halted the early added therapeutic effect. Conclusion: Overall, we demonstrated that 3D µCT/BLT is a valuable non-isotope-based technology for whole-body cell therapy monitoring and investigating CAR T cell pharmacokinetics. We also presented combining LSFM and MICS for ex vivo 3D- and 2D-microscopy tissue analysis to assess intratumoral therapeutic cell distribution and status

Neutrophil Extracellular Traps Contain Calprotectin, a Cytosolic Protein Complex Involved in Host Defense against Candida albicans

Neutrophils are the first line of defense at the site of an infection. They encounter and kill microbes intracellularly upon phagocytosis or extracellularly by degranulation of antimicrobial proteins and the release of Neutrophil Extracellular Traps (NETs). NETs were shown to ensnare and kill microbes. However, their complete protein composition and the antimicrobial mechanism are not well understood. Using a proteomic approach, we identified 24 NET-associated proteins. Quantitative analysis of these proteins and high resolution electron microscopy showed that NETs consist of modified nucleosomes and a stringent selection of other proteins. In contrast to previous results, we found several NET proteins that are cytoplasmic in unstimulated neutrophils. We demonstrated that of those proteins, the antimicrobial heterodimer calprotectin is released in NETs as the major antifungal component. Absence of calprotectin in NETs resulted in complete loss of antifungal activity in vitro. Analysis of three different Candida albicans in vivo infection models indicated that NET formation is a hitherto unrecognized route of calprotectin release. By comparing wild-type and calprotectin-deficient animals we found that calprotectin is crucial for the clearance of infection. Taken together, the present investigations confirmed the antifungal activity of calprotectin in vitro and, moreover, demonstrated that it contributes to effective host defense against C. albicans in vivo. We showed for the first time that a proportion of calprotectin is bound to NETs in vitro and in vivo

Assembling proteomics data as a prerequisite for the analysis of large scale experiments

<p>Abstract</p> <p>Background</p> <p>Despite the complete determination of the genome sequence of a huge number of bacteria, their proteomes remain relatively poorly defined. Beside new methods to increase the number of identified proteins new database applications are necessary to store and present results of large- scale proteomics experiments.</p> <p>Results</p> <p>In the present study, a database concept has been developed to address these issues and to offer complete information via a web interface. In our concept, the Oracle based data repository system SQL-LIMS plays the central role in the proteomics workflow and was applied to the proteomes of <it>Mycobacterium tuberculosis</it>, <it>Helicobacter pylori</it>, <it>Salmonella typhimurium </it>and protein complexes such as 20S proteasome. Technical operations of our proteomics labs were used as the standard for SQL-LIMS template creation. By means of a Java based data parser, post-processed data of different approaches, such as LC/ESI-MS, MALDI-MS and 2-D gel electrophoresis (2-DE), were stored in SQL-LIMS. A minimum set of the proteomics data were transferred in our public 2D-PAGE database using a Java based interface (Data Transfer Tool) with the requirements of the PEDRo standardization. Furthermore, the stored proteomics data were extractable out of SQL-LIMS via XML.</p> <p>Conclusion</p> <p>The Oracle based data repository system SQL-LIMS played the central role in the proteomics workflow concept. Technical operations of our proteomics labs were used as standards for SQL-LIMS templates. Using a Java based parser, post-processed data of different approaches such as LC/ESI-MS, MALDI-MS and 1-DE and 2-DE were stored in SQL-LIMS. Thus, unique data formats of different instruments were unified and stored in SQL-LIMS tables. Moreover, a unique submission identifier allowed fast access to all experimental data. This was the main advantage compared to multi software solutions, especially if personnel fluctuations are high. Moreover, large scale and high-throughput experiments must be managed in a comprehensive repository system such as SQL-LIMS, to query results in a systematic manner. On the other hand, these database systems are expensive and require at least one full time administrator and specialized lab manager. Moreover, the high technical dynamics in proteomics may cause problems to adjust new data formats. To summarize, SQL-LIMS met the requirements of proteomics data handling especially in skilled processes such as gel-electrophoresis or mass spectrometry and fulfilled the PSI standardization criteria. The data transfer into a public domain via DTT facilitated validation of proteomics data. Additionally, evaluation of mass spectra by post-processing using MS-Screener improved the reliability of mass analysis and prevented storage of data junk.</p

Reorientation of Spin Density Waves in Cr(001) Films induced by Fe(001) Cap Layers

Proximity effects of 20 \AA thin Fe layers on the spin density waves (SDWs) in epitaxial Cr(001) films are revealed by neutron scattering. Unlike in bulk Cr we observe a SDW with its wave vector Q pointing along only one {100} direction which depends dramatically on the film thickness t_{Cr}. For t_{Cr} < 250 \AA the SDW propagates out-of-plane with the spins in the film plane. For t_{Cr} > 1000 \AA the SDW propagates in the film plane with the spins out-of-plane perpendicular to the in-plane Fe moments. This reorientation transition is explained by frustration effects in the antiferromagnetic interaction between Fe and Cr across the Fe/Cr interface due to steps at the interface.Comment: 4 pages (RevTeX), 3 figures (EPS