Evidence for P-Glycoprotein Involvement in Cell Volume Regulation Using Coulter Sizing in Flow Cytometry

The regulation of cell volume is an essential function that is coupled to a variety of physiological processes such as receptor recycling, excitability and contraction, cell proliferation, migration, and programmed cell death. Under stress, cells undergo emergency swelling and respond to such a phenomenon with a regulatory volume decrease (RVD) where they release cellular ions, and other osmolytes as well as a concomitant loss of water. The link between P-glycoprotein, a transmembrane transporter, and cell volume regulation is controversial, and changes in cells volume are measured using microscopy or electrophysiology. For instance, by using the patch-clamp method, our team demonstrated that chloride currents activated in the RVD were more intense and rapid in a breast cancer cell line overexpressing the P-glycoprotein (P-gp). The Cell Lab Quanta SC is a flow cytometry system that simultaneously measures electronic volume, side scatter and three fluorescent colors; altogether this provides unsurpassed population resolution and accurate cell counting. Therefore, here we propose a novel method to follow cellular volume. By using the Coulter-type channel of the cytometer Cell Lab Quanta SC MPL (multi-platform loading), we demonstrated a role for the P-gp during different osmotic treatments, but also a differential activity of the P-gp through the cell cycle. Altogether, our data strongly suggests a role of P-gp in cell volume regulation.     

Dark matter searches with H.E.S.S.

WIMP annihilations are required to reproduce the correct dark matter abundance in the Universe. This process can occur in dense regions of our Galaxy such as the Galactic center, dwarf galaxies and other types of sub-haloes. High-energy γ-rays are produced in dark matter particle collisions and can be detected by Cherenkov telescopes such as H.E.S.S. We report here the search for γ-ray signals from the Galactic center and the nearby Sagittarius and Canis Major satellites. In the absence of a convincing signal, modelling the dark matter density within these objects allows to put constraints on the WIMP parameters such as its mass and annihilation cross-section. Beyond these targeted searches, the wide-field survey of the inner Galaxy with H.E.S.S. is used to constrain sub-halo formation models involving intermediate-mass black holes.     

Degradation of semiconducting polymers by concentrated sunlight

A lens based sunlight concentration setup was used to accelerate the degradation of semiconducting polymers. Sunlight was collected outdoor and focused into an optical fiber bundle allowing for indoor experimental work. Photo-degradation of several polymers was studied by UV-vis absorbance spectroscopy and infra-red spectroscopy. This showed that the degradation rate is significantly increased by increasing illumination intensity. Acceleration factors exceeding 100 compared to standard 1 sun illumination were observed for solar concentration of 200 suns in the case of P3HT. A comparison between infra-red spectra of MEH-PPV degraded at 1 sun intensity and at high solar concentration only showed minor deviations in degradation mechanisms. The acceleration factor was found to vary linearly with the solar concentration. Finally, a comparison of the degradation rates at 1 sun and 100 suns was carried out in a materials study employing five different conjugated polymers relevant to polymer solar cells for which acceleration factors in the range 19-55 were obtained.     

TP53 Targeted Deep Sequencing of Cell-Free DNA in Esophageal Squamous Cell Carcinoma Using Low-Quality Serum: Concordance with Tumor Mutation

Circulating cell-free DNA (cfDNA) is emerging as a potential tumor biomarker. CfDNA-based biomarkers may be applicable in tumors without an available non-invasive screening method among at-risk populations. Esophageal squamous cell carcinoma (ESCC) and residents of the Asian cancer belt are examples of those malignancies and populations. Previous epidemiological studies using cfDNA have pointed to the need for high volumes of good quality plasma (i.e., >1 mL plasma with 0 or 1 cycles of freeze-thaw) rather than archival serum, which is often the main available source of cfDNA in retrospective studies. Here, we have investigated the concordance of TP53 mutations in tumor tissue and cfDNA extracted from archival serum left-over from 42 cases and 39 matched controls (age, gender, residence) in a high-risk area of Northern Iran (Golestan). Deep sequencing of TP53 coding regions was complemented with a specialized variant caller (Needlestack). Overall, 23% to 31% of mutations were concordantly detected in tumor and serum cfDNA (based on two false discovery rate thresholds). Concordance was positively correlated with high cfDNA concentration, smoking history (p-value = 0.02) and mutations with a high potential of neoantigen formation (OR; 95%CI = 1.9 (1.11–3.29)), suggesting that tumor DNA release in the bloodstream might reflect the effects of immune and inflammatory context on tumor cell turnover. We identified TP53 mutations in five controls, one of whom was subsequently diagnosed with ESCC. Overall, the results showed that cfDNA mutations can be reliably identified by deep sequencing of archival serum, with a rate of success comparable to plasma. Nonetheless, 70% non-identifiable mutations among cancer patients and 12% mutation detection in controls are the main challenges in applying cfDNA to detect tumor-related variants when blindly targeting whole coding regions of the TP53 gene in ESCC.     

The Conversion of UDP-Glc to UDP-Man: In Silico and Biochemical Exploration To Improve the Catalytic Efficiency of CDP-Tyvelose C2-Epimerases

A promiscuous CDP-tyvelose 2-epimerase (TyvE) from Thermodesulfatator atlanticus (TaTyvE) belonging to the nucleotide sugar active short-chain dehydrogenase/reductase superfamily (NS-SDRs) was recently discovered. TaTyvE performs the slow conversion of NDP-glucose (NDP-Glc) to NDP-mannose (NDP-Man). Here, we present the sequence fingerprints that are indicative of the conversion of UDP-Glc to UDP-Man in TyvE-like enzymes based on the heptagonal box motifs. Our data-mining approach led to the identification of 11 additional TyvE-like enzymes for the conversion of UDP-Glc to UDP-Man. We characterized the top two wild-type candidates, which show a 15- and 20-fold improved catalytic efficiency, respectively, on UDP-Glc compared to TaTyvE. In addition, we present a quadruple variant of one of the identified enzymes with a 70-fold improved catalytic efficiency on UDP-Glc compared to TaTyvE. These findings could help the design of new nucleotide production pathways starting from a cheap sugar substrate like glucose or sucrose.     

A classification approach to efficient global optimization in presence of non-computable domains

Gaussian-Process based optimization methods have become very popular in recent years for the global optimization of complex systems with high computational costs. These methods rely on the sequential construction of a statistical surrogate model, using a training set of computed objective function values, which is refined according to a prescribed infilling strategy. However, this sequential optimization procedure can stop prematurely if the objective function cannot be computed at a proposed point. Such a situation can occur when the search space encompasses design points corresponding to an unphysical configuration, an ill-posed problem, or a non-computable problem due to the limitation of numerical solvers. To avoid such a premature stop in the optimization procedure, we propose to use a classification model to learn non-computable areas and to adapt the infilling strategy accordingly. Specifically, the proposed method splits the training set into two subsets composed of computable and non-computable points. A surrogate model for the objective function is built using the training set of computable points, only, whereas a probabilistic classification model is built using the union of the computable and non-computable training sets. The classifier is then incorporated in the surrogate-based optimization procedure to avoid proposing new points in the non-computable domain while improving the classification uncertainty if needed. The method has the advantage to automatically adapt both the surrogate of the objective function and the classifier during the iterative optimization process. Therefore, non-computable areas do not need to be a priori known. The proposed method is applied to several analytical problems presenting different types of difficulty, and to the optimization of a fully nonlinear fluid-structure interaction system. The latter problem concerns the drag minimization of a flexible hydrofoil with cavitation constraints. The efficiency of the proposed method compared favorably to a reference evolutionary algorithm, except for situations where the feasible domain is a small portion of the design space.     

Effects of forearm and palm supports on the upper extremity during computer mouse use

The use of forearm and palm supports has been associated with lower neck and shoulder muscle activity as well as reduced musculoskeletal discomfort during keyboard use, however, few studies have investigated their effect during computer mouse use. Eight men and eight women completed several computer mousing tasks in six arm support conditions: Forearm Support, Flat Palm Support, Raised Palm Support, Forearm + Flat Palm Support, Forearm + Raised Palm Support, and No Support. Concurrently, an infrared three-dimensional motion analysis system measured postures, six-degree-of-freedom force-torque sensors measured applied forces & torques, and surface electromyography measured muscle activity. The use of forearm support compared to the no support condition was significantly associated with less shoulder muscle activity & torque, and the raised palm support was associated with less wrist extension. Forearm supports reduced shoulder flexion torque by 90% compared to no support. The use of either support also resulted in lower applied forces to the mouse pad. Participants reported less musculoskeletal discomfort when using a support. These results provide recommendations for office workstation setup and inform ergonomists of effective ways to reduce musculoskeletal exposures.     

Influence of elastomer distribution on the cryogenic microcracking of carbon fiber/epoxy composites

Carbon fiber/epoxy laminates containing three different types of rubber modifiers, separately and in combination, were developed for testing in a cryogenic environment. Preformed rubber particles, core shell rubber, and solid carboxyl‐functionalized rubber were chosen as additives to a model prepreg matrix to control the placement of the rubber within the resulting laminates. Cryogenic microcracking and mode I and II fracture toughness and interlaminar shear strength experiments were performed. Scanning electron microscopy was used to observe fracture surfaces of the rubber‐modified laminates. Fracture toughness properties were improved while the ILSS decreased because of the presence of these rubber modifiers. It was observed that the presence of these modifiers significantly reduced the microcrack density of the laminates exposed to cryogenic cycling, and in the case of one, even eliminated microcracking entirely. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 2268–2275, 2003