Yields and product comparison between Escherichia coli BL21 and W3110 in industrially relevant conditions: anti-c-Met scFv as a case study.

Introduction: In the biopharmaceutical industry, Escherichia coli is one of the preferred expression hosts for large-scale production of therapeutic proteins. Although increasing the product yield is important, product quality is a major factor in this industry because greatest productivity does not always correspond with the highest quality of the produced protein. While some post-translational modifications, such as disulphide bonds, are required to achieve the biologically active conformation, others may have a negative impact on the product's activity, effectiveness, and/or safety.

Superparamagnetic nanoparticles with LC polymer brush shell as efficient dopants for ferronematic phases.

Liquid crystal (LC) based magnetic materials consisting of LC hosts doped with functional magnetic nanoparticles enable optical switching of the mesogens at moderate magnetic field strengths and thereby open the pathway for the design of novel smart devices. A promising route for the fabrication of stable ferronematic phases is the attachment of a covalently bound LC polymer shell onto the surface of nanoparticles. With this approach, ferronematic phases based on magnetically blocked particles and the commercial LC 4-cyano-4'-pentylbiphenyl (5CB) liquid crystal were shown to have a sufficient magnetic sensitivity, but the mechanism of the magneto-nematic coupling is unidentified.

Synthesis of coagulation factors during long-term ex situ liver perfusion.

Robust viability assessment of grafts during normothermic liver perfusion is a prerequisite for organ use. Coagulation parameters are used commonly for liver assessment in patients. However, they are not yet included in viability assessment during ex situ perfusion.

Creation of a PDMS Polymer Brush on SiO-Based Nanoparticles by Surface-Initiated Ring-Opening Polymerization.

The incorporation of nanoparticles into soft matrices opens a broad spectrum of novel property combinations. However, one of the major challenges for these systems remains the compatibilization of particles with the surrounding matrix by proper surface functionalization. For silicon-based systems or liquid crystalline phases, polydimethylsiloxane (PDMS) brushes at the surface of particles increase the stability against particle agglomeration in such systems.

Efficient ferronematic coupling with polymer-brush particles.

Switching of liquid crystal phases is of enormous technological importance and enables digital displays, thermometers and sensors. As an alternative to electric fields or temperature, magnetic fields are an interesting trigger, as they are on the one hand versatile to design, and on the other hand, they are compatible with a bouquet of applications. An interesting option to enable the magnetic switchability of nematic phases is by doping them with functional magnetic nanoparticles, but it remains a challenge to achieve well-compatibilized and stable ferronematic phases.

Evaluation of N8-GP Activity Using a One-Stage Clotting Assay: A Single-Center Experience.

N8-GP (ESPEROCT®; turoctocog alfa pegol; Novo Nordisk A/S, Bagsvaerd, Denmark) is an extended half-life recombinant factor VIII (FVIII) molecule. FVIII-deficient plasma spiked with N8-GP can be accurately measured using many activated partial thromboplastin time (aPTT)-based one-stage clotting assay reagents with normal human plasma calibrators. To date, there are few data on the measurement accuracy of samples from patients treated with N8-GP.

The role of structural anisotropy in the magnetooptical response of an organoferrogel with mobile magnetic nanoparticles.

We investigate the structure and the magnetooptical response of isotropic and anisotropic fibrillous organoferrogels with mobile magnetic nanoparticles (MNPs). We demonstrate that the presence of the gel network restricts the magnetooptical response of the ferrogel. Even though the ferrogel exhibits no magnetic hysteresis, an optical hysteresis has been found.

Oxidative stress-induced structural changes in the microtubule-associated flavoenzyme Irc15p from Saccharomyces cerevisiae.

The genome of the yeast Saccharomyces cerevisiae encodes a canonical lipoamide dehydrogenase (Lpd1p) as part of the pyruvate dehydrogenase complex and a highly similar protein termed Irc15p (increased recombination centers 15). In contrast to Lpd1p, Irc15p lacks a pair of redox active cysteine residues required for the reduction of lipoamide and thus it is very unlikely that Irc15p performs a similar dithiol-disulfide exchange reaction as reported for lipoamide dehydrogenases. We expressed IRC15 in Escherichia coli and purified the produced protein to conduct a detailed biochemical characterization.

Cognitive and Behavioral Consequences of Pediatric Delirium: A Pilot Study.

Objectives: To investigate the long-term impact of postoperative delirium in children.

Design: Single-center point prevalence study.

Setting: Twenty-two bed PICU.

Structure, biochemical and kinetic properties of recombinant Pst2p from Saccharomyces cerevisiae, a FMN-dependent NAD(P)H:quinone oxidoreductase.

The genome of the yeast Saccharomyces cerevisiae encodes four flavodoxin-like proteins, namely Lot6p, Pst2p, Rfs1p and Ycp4p. Thus far only Lot6p was characterized in detail demonstrating that the enzyme possesses NAD(P)H:quinone oxidoreductase activity. In the present study, we heterologously expressed PST2 in Escherichia coli and purified the produced protein to conduct a detailed biochemical and structural characterization.

Catalytic competence, structure and stability of the cancer-associated R139W variant of the human NAD(P)H:quinone oxidoreductase 1 (NQO1).

The human NAD(P)H:quinone oxidoreductase 1 (NQO1; EC1.6.99.

Mosaic deletion of EXOC6B: further evidence for an important role of the exocyst complex in the pathogenesis of intellectual disability.

We describe a boy with developmental delay, speech delay, and minor dysmorphic features with a heterozygous de novo ∼460 kb deletion at 2p13.2 involving only parts of EXOC6B present in about 50% of lymphocytes. This widely expressed gene encodes the exocyst component 6B, which is part of a multiprotein complex required for targeted exocytosis.

Nephroblastomas show low expression of microR-204 and high expression of its target, the oncogenic transcription factor MEIS1.

By comparing several studies we identified a possible deregulation of the transcription factors PBX2 (pre-B-cell leukemia homeobox 2) and one of its binding partners, MEIS1 (Meis homeobox 1) in nephroblastomas. The regulation of MEIS1 is complex, and its expression is known to be influenced by changes of promoter methylation and binding of microRNA-204 (miR-204). Therefore, in our study, we assessed the expression of MEIS1 and PBX2 and the factors regulating expression of MEIS1 in nephroblastomas.

The flavoproteome of the yeast Saccharomyces cerevisiae.

Genome analysis of the yeast Saccharomyces cerevisiae identified 68 genes encoding flavin-dependent proteins (1.1% of protein encoding genes) to which 47 distinct biochemical functions were assigned. The majority of flavoproteins operate in mitochondria where they participate in redox processes revolving around the transfer of electrons to the electron transport chain.

A 15q24 microdeletion in transient myeloproliferative disease (TMD) and acute megakaryoblastic leukaemia (AMKL) implicates PML and SUMO3 in the leukaemogenesis of TMD/AMKL.

Transient myeloproliferative disorder (TMD) of the newborn and acute megakaryoblastic leukaemia (AMKL) in children with Down syndrome (DS) represent paradigmatic models of leukaemogenesis. Chromosome 21 gene dosage effects and truncating mutations of the X-chromosomal transcription factor GATA1 synergize to trigger TMD and AMKL in most patients. Here, we report the occurrence of TMD, which spontaneously remitted and later progressed to AMKL in a patient without DS but with a distinct dysmorphic syndrome.

Percutaneous osteoplasty as a treatment for painful malignant bone lesions of the pelvis and femur.

The purposes of this report are to describe percutaneous osteoplasty as a highly effective minimally invasive procedure to treat painful malignant bone lesions of the pelvis, ilium, and femur and to discuss the relevant literature. Five patients with histologically proven metastases to the pelvis, ilium, or femur were treated by percutaneous injection of liquid bone cement as an attempt to control severe bone pain. After percutaneous osteoplasty, all five patients experienced immediate and substantial pain relief and did not require pain medication for the duration of follow-up.

Interaction of Gbeta3s, a splice variant of the G-protein Gbeta3, with Ggamma- and Galpha-proteins.

The T-allele of a polymorphism (C825T) in the gene of the G-protein beta3-subunit is associated with a complex phenotype (hypertension, obesity, altered drug responses) and the occurrence of a splice variant termed Gbeta3s which lacks one of the seven WD-domains that compose Gbeta-proteins. Here, we analysed Gbetagamma dimer formation and Galpha activation by Gbeta3s, key functional characteristics of Gbeta-proteins. Cleavage protection assays frequently used to analyse Gbeta1gamma and Gbeta2gamma dimer formation failed for Gbeta3 and Gbeta3s, while in coprecipitation assays, dimerization of Gbeta3 and Gbeta3s with Ggamma5, Ggamma8(c) and Ggamma12 could be demonstrated.