Next generation adoptive immunotherapy--human T cells as carriers of therapeutic nanoparticles

An important step in adoptive immunotherapy in general and specifically with respect to cancer treatment is the initiation of an inflammatory T cell response at the tumor site. Here we suggest a new concept for a controlled inflammatory response in which the intrinsic cytotoxic properties of T cells are upgraded with the properties of nanoparticles transfected into the T cells during the ex vivo expansion process. We report in vitro upgrading of human T cells using PEGylated boron carbide nanoparticles functionalised with a translocation peptide aimed at Boron Neutron Capture Therapy (BNCT). A key finding is that the metabolism of such upgraded human T cells were not affected by a payload of 0.13 pg boron per cell and that the nanoparticles were retained in the cell population after several cell divisions. This is vital for transporting nanoparticles by T cells to the tumor site.     

An analysis of the tensile elongation to failure of laminated metal composites in the presence of strain-rate hardening

We extend the analysis of Hutchinson and Neale (Hutchinson JW, Neale KW. Acta Metall 1977;25:839) and Ghosh (Ghosh AK. Acta Metall 1977;25:1413) predicting the tensile elongation to failure of strain-rate-dependent plastic materials to two-phase composites deforming quasistatically according to the equistrain rule of mixtures. The analysis incorporates the influence of work hardening and strain-rate hardening in both composite constituent phases. It is shown that the problem can be formulated in a manner that condenses the seven underlying material parameters into four dimensionless numbers for composites of power-law hardening phases, the number of parameters falling to two for linear hardening. It then emerges that the stabilizing influence of both work hardening and strain-rate hardening is, within assumptions of the model, always predominantly exerted by the phase that carries the greater share of the composite stress. It is also shown that the prediction can be simplified so as to enable an approximate but convenient direct graphical deduction of the tensile elongation of ductile laminated metal composites (LMCs), knowing the work hardening and strain-rate hardening characteristics of the two phases making the composite. The utility of this graphical scheme is illustrated with two examples, namely LMCs containing one phase of (moderately ductile) aluminium alloy or of a (low-ductility) nanocrystalline metal.     

Wafer scale imprint uniformity evaluated by LSPR spectroscopy: a high volume characterization method for nanometer scale structures

We exploit the localized surface-plasmon resonance (LSPR) of terahertz gold gammadion structures for wafer scale critical dimension metrology of nanostructures. The proposed characterization method, LSPR spectroscopy, is based on optical transmission measurements and is benchmarked against numerical simulations of imprinted structures characterized by atomic force microscopy. There is a fair agreement between the two methods and the simulations enable the translation of optical spectra to critical dimensions of the physical structures, a concept known from scatterometry. The results demonstrate the potential of LSPR spectroscopy as an alternative characterization method to scanning electron microscopy, atomic force microscopy and scatterometry.     

Numerical analysis of transport phenomena for designing of ejector in PEM forklift system

In the present study, Computational Fluid Dynamics (CFD) technique is used to design an ejector for anode recirculation in an automotive PEMFC system. A CFD model is firstly established and tested against well-documented and relevant solutions from the literature, and then used for different ejector geometries under different operating conditions. Results showed that a single ejector with optimized geometry cannot cover the required recirculation in the entire range of the fuel cell current. Having two ejectors for different ranges of currents is thus proposed as an alternative solution in which the system can better take the advantage of ejectors for recirculation purpose. In addition, the operating mode of one variable nozzle ejector has been investigated and compared with aforementioned cases. The results showed that the variable nozzle ejector can work in the same operational mode as in the case with two ejectors. However, in practice, the latter one needs a more complicated control system and it is more difficult to manufacture.     

Effect of pre-slaughter physiological conditions on the oxidative stability of colour and lipid during chill storage of pork

The physiological condition of the live animal was found to significantly affect colour, lipid oxidation and water holding capacity of chill stored pork chops (M. Longissimus dorsi) in a study, where various pre-slaughter conditions were achieved by the following four treatments: (A) control; (B) subjected to treadmill exercise immediately prior to stunning; (C) given epinephrine injection 15 h prior to slaughter; and (D) given epinephrine injection 15 h before slaughter and further subjected to treadmill exercise immediately before stunning. The treatments resulted in variations in energy metabolites (glycogen, lactate, creatine phosphate, ATP) and ultimate pH (pH_u), with the lowest pH_u in chops from treatments A and B, and in significantly different tristimulus colour L^*-, a^*- and b^*-parameters, although the effect of treatment on colour was not consistent during the chill storage period of 6 days. Overall, chops from treatments A and B had significantly higher L^*- and b^*-values (were paler and less blue) than chops from C and D during storage under conditions typical for retail trade. The initial a^*-values were higher (redder) in chops from treatments A and B, but the colour, as judged by the a^*-values, was less stable in meat from these treatments compared with treatments C and D. Lipid oxidation, evaluated by thiobarbituric acid reactive substances (TBARS) in the fresh meat, and drip loss, measured after 6 days of storage, were both significantly higher in chops from treatments A and B compared to chops obtained from treatments C and D. Statistical analysis relating the pH and the level of various energy metabolites post-mortem in the individual animals to the measured quality parameters, revealed that pH_u was the most important factor affecting product quality. In conclusion, over all product quality depends on obtaining a pH_u in the narrow range where both meat quality parameters such as colour, lipid oxidation and drip loss as well as microbiological aspects have to be considered.     

Considerations on Probe Design for Affinity-Guided Protein Conjugation

The plethora of methods developed for the creation of protein conjugates often differs significantly with regard to the heterogeneity of the resulting products, in the degree of genetic manipulation of the protein required, and in the technical skills required to perform the conjugation procedure. Affinity-guided protein conjugation is a protein labeling methodology based on noncovalent binding interactions between a labeling probe and the protein of interest. These interactions increase the local concentration of a reactive group in the probe on the protein surface thus facilitating the conjugation in proximity of the complexation site. The ability to produce high-quality conjugates from nongenetically modified proteins both in vitro, but also in cells, demonstrates the power of affinity-guided protein conjugation. Here, we present the progress of affinity-guided protein conjugation in relation to selective protein labeling in living systems and the formation of high-quality protein conjugates. Furthermore, the probe design will be discussed in relation to the utility of the probe for labeling in vitro or in living systems.     

Thermal modeling and temperature control of a PEM fuel cell system for forklift applications

Temperature changes in PEM fuel cell stacks are considerably higher during load variations and have a negative impact as they generate thermal stresses and stack degradation. Cell hydration is also of vital importance in fuel cells and it is strongly dependent on operating temperature. A combination of high temperature and reduced humidity increases the degradation rate. Stack thermal management and control are, thus, crucial issues in PEM fuel cell systems especially in automotive applications such as forklifts.     

Contrast-medium-induced ventricular fibrillation: arrhythmogenic mechanisms and the role of antiarrhythmic drugs in dogs

RATIONALE AND OBJECTIVES: Small electrolyte additions to a nonionic contrast medium reduce the risk of ventricular fibrillation (VF) during wedged catheter injection of a contrast medium. The current study was designed to further investigate contrast-medium-induced VF by studying the effect of pretreatment with different antiarrhythmic drugs. METHODS: During a simulated wedged catheter situation, iohexol was injected into the anterior descending branch of the left coronary artery in five open-chest, anesthetized dogs pretreated with lidocaine, propranolol, amiodarone, almokalant, or verapamil. RESULTS: Wedging the catheter for 60 sec did not induce VF. However, all 15 wedged catheter injections with iohexol induced VF within 28 sec (19 +/- 1 [mean +/- standard error of the mean]) despite pretreatment with antiarrhythmic drugs. Prior to VF, conduction was slowed and monophasic action potential duration lengthened in the contrast-medium-perfused myocardium, although no significant changes occurred in the control area. CONCLUSION: The combination of catheter wedging and long-lasting contrast medium injection has a high risk of causing VF. Although adding a small amount of electrolytes to nonionic contrast media can reduce the risk of VF, antiarrhythmic drug therapy may not have a protective effect.     

A six-domain structural model for Escherichia coli translation initiation factor IF2. Characterisation of twelve surface epitopes

The Escherichia coli translation initiation factor IF2 is a 97 kDa protein which interacts with the initiator fMet-tRNAfMet, GTP and the ribosomal subunits during initiation of protein biosynthesis. For structural and functional investigations of the factor, we have raised and characterised monoclonal antibodies against E. coli IF2. Twelve epitopes have been localised at the surface of the protein molecule by three different methods: Interactions of the monoclonal antibodies with nested deletion mutants of IF2, comparison of the relative location of the epitopes in a competition immunoassay and cross-reactivity analyses of the monoclonal antibodies towards IF2 from Salmonella typhimurium, Klebsiella oxytoca, Enterobacter cloacae, Proteus vulgaris, and Bacillus stearothermophilus. These data are combined with predicted secondary structure and discussed in relation to a six-domain structural model for IF2. The model describes IF2 as a slightly elongated molecule with a structurally compact C-terminal domain, a well-conserved central GTP-binding domain, and a highly charged, solvent exposed N-terminal with protruding alpha-helical structures.