Characterization of anticoagulant heparinoids by immunoprofiling

Heparinoids are used in the clinic as anticoagulants. A specific pentasaccharide in heparinoids activates antithrombin III, resulting in inactivation of factor Xa and–when additional saccharides are present–inactivation of factor IIa. Structural and functional analysis of the heterogeneous heparinoids generally requires advanced equipment, is time consuming, and needs (extensive) sample preparation. In this study, a novel and fast method for the characterization of heparinoids is introduced based on reactivity with nine unique anti-heparin antibodies. Eight heparinoids were biochemically analyzed by electrophoresis and their reactivity with domain-specific anti-heparin antibodies was established by ELISA. Each heparinoid displayed a distinct immunoprofile matching its structural characteristics. The immunoprofile could also be linked to biological characteristics, such as the anti-Xa/anti-IIa ratio, which was reflected by reactivity of the heparinoids with antibodies HS4C3 (indicative for 3-O-sulfates) and HS4E4 (indicative for domains allowing anti-factor IIa activity). In addition, the immunoprofile could be indicative for heparinoid-induced side-effects, such as heparin-induced thrombocytopenia, as illustrated by reactivity with antibody NS4F5, which defines a very high sulfated domain. In conclusion, immunoprofiling provides a novel, fast, and simple methodology for the characterization of heparinoids, and allows high-throughput screening of (new) heparinoids for defined structural and biological characteristics

Measurement of the longitudinal diffusion of ionization electrons in the MicroBooNE detector

Abstract: Accurate knowledge of electron transport properties is vital to understanding the information provided by liquid argon time projection chambers (LArTPCs). Ionization electron drift-lifetime, local electric field distortions caused by positive ion accumulation, and electron diffusion can all significantly impact the measured signal waveforms. This paper presents a measurement of the effective longitudinal electron diffusion coefficient, DL, in MicroBooNE at the nominal electric field strength of 273.9 V/cm. Historically, this measurement has been made in LArTPC prototype detectors. This represents the first measurement in a large-scale (85 tonne active volume) LArTPC operating in a neutrino beam. This is the largest dataset ever used for this measurement. Using a sample of ∼70,000 through-going cosmic ray muon tracks tagged with MicroBooNE's cosmic ray tagger system, we measure DL = 3.74+0.28 -0.29 cm2/s

The RET/PTC3 oncogene activates classical NF-[kappa]B by stabilizing NIK

The oncogenic fusion protein RET/PTC3 (RP3) that is expressed in papillary thyroid carcinoma (PTC) and thyroid epithelia in Hashimoto's thyroiditis activates nuclear factor-kappa B (NF-[kappa]B) and induces pro-inflammatory gene expression; however, the mechanism of this activation is unknown. To address this, we expressed RP3 in murine embryonic fibroblasts (MEFs) lacking key classical and noncanonical NF-[kappa]B signaling components. In wild-type MEFs, RP3 upregulated CCL2, CXCL1, granulocyte-macrophage colony-stimulating factor and tumor necrosis factor expression and activated classical but not noncanonical NF-[kappa]B. RP3-activated NF-[kappa]B-in I[kappa]B kinase (IKK)[beta].sup.-/-] MEFs but not IKK[alpha]- or NF-[kappa]B essential modulator (NEMO)-deficient cells and activation was inhibited by a peptide that blocks NEMO binding to the IKKs. RP3 increased the levels of NF-[kappa]B-inducing kinase (NIK) and did not activate NF-[kappa]B in NIK-deficient MEFs. Notably, NIK stabilization was not accompanied by TRAF3 degradation demonstrating that RP3 disrupts normal basal NIK regulation. Dominant-negative NIK blocked RP3-induced NF-[kappa]B activation and an RP3 signaling mutant ([RP3.SUP.Y588F]) did not stabilize NIK. Finally, examination of PTC specimens revealed strong positive staining for NIK. We therefore conclude that RP3 activates classical NF-[kappa]B via NIK, NEMO and IKK[alpha]. Importantly, our findings reveal a novel mechanism for oncogene-induced NF-[kappa]B activation via stabilization of NIK. Oncogene (2011) 30, 87-96; doi: 10.1038/onc.2010.396; published online 6 September 2010 Keywords: Hashimoto's thyroiditis; IKK; NIK; NF-[kappa]B; RET/PTC; thyroid cancerAcademi

Individualised antibiotic dosing for patients who are critically ill: challenges and potential solutions

Item does not contain fulltextInfections in critically ill patients are associated with persistently poor clinical outcomes. These patients have severely altered and variable antibiotic pharmacokinetics and are infected by less susceptible pathogens. Antibiotic dosing that does not account for these features is likely to result in suboptimum outcomes. In this Review, we explore the challenges related to patients and pathogens that contribute to inadequate antibiotic dosing and discuss how to implement a process for individualised antibiotic therapy that increases the accuracy of dosing and optimises care for critically ill patients. To improve antibiotic dosing, any physiological changes in patients that could alter antibiotic concentrations should first be established; such changes include altered fluid status, changes in serum albumin concentrations and renal and hepatic function, and microvascular failure. Second, antibiotic susceptibility of pathogens should be confirmed with microbiological techniques. Data for bacterial susceptibility could then be combined with measured data for antibiotic concentrations (when available) in clinical dosing software, which uses pharmacokinetic/pharmacodynamic derived models from critically ill patients to predict accurately the dosing needs for individual patients. Individualisation of dosing could optimise antibiotic exposure and maximise effectiveness

Frequency doubling of multi-terawatt picosecond pulses

International audienceThe frequency-doubling efficiency and resultant focal spot quality of a large aperture (140 × 89 mm) subpicosecond, chirped pulse amplified (CPA) 1054-nm beam for laser–matter interaction studies has been investigated using the Vulcan Nd:glass laser system (Danson et al. 1998). The effect of B-integral on the CPA beam quality was studied and is shown not to be the dominant cause of the observed frequency-doubled beam break-up. Conversion efficiency tests were carried out on small aperture KDP (type 1) crystals at a range of incident intensities up to 3 × 1011 W/cm2 giving the optimum crystal thickness for pulses in the 0.3–3 ps region. A large-aperture frequency-doubled beam was commissioned and delivered pulses of over 10 TW onto target for an electron acceleration experiment