Prerequisites for cytokine measurements in clinical trials with multiplex immunoassays

<p>Abstract</p> <p>Background</p> <p>Growing knowledge about cellular interactions in the immune system, including the central role of cytokine networks, has lead to new treatments using monoclonal antibodies that block specific components of the immune system. Systemic cytokine concentrations can serve as surrogate outcome parameters of these interventions to study inflammatory pathways operative in patients <it>in vivo</it>. This is now possible due to novel technologies such as multiplex immunoassays (MIA) that allows detection of multiple cytokines in a single sample. However, apparently trivial underappreciated processes, (sample handling and storage, interference of endogenous plasma proteins) can greatly impact the reliability and reproducibility of cytokine detection.</p> <p>Therefore we set out to investigate several processes that might impact cytokine profiles such as blood collecting tubes, duration of storage, and number of freeze thawing cycles.</p> <p>Results</p> <p>Since under physiological conditions cytokine concentrations normally are low or undetectable we spiked cytokines in the various plasma and serum samples. Overall recoveries ranged between 80-120%. Long time storage showed cytokines are stable for a period up to 2 years of storage at -80°C. After 4 years several cytokines (IL-1α, IL-1β, IL-10, IL-15 and CXCL8) degraded up to 75% or less of baseline values. Furthermore we show that only 2 out of 15 cytokines remained stable after several freeze-thawing cycles. We also demonstrate implementation of an internal control for multiplex cytokine immunoassays.</p> <p>Conclusion</p> <p>All together we show parameters which are essential for measurement of cytokines in the context of clinical trials.</p

Helical microstructures in molluscan biomineralization are a biological example of close packed helices that may form from a colloidal liquid crystal precursor in a twist–bend nematic phase

We demonstrate that nature has produced a close-packed helical twisted filamentous material in the biomineralization of the mollusk. In liquid crystals, twist–bend nematics have been predicted and observed. We present and analyze evidence that the helical biomineral microstructure of mollusk shells may be formed from such a liquid-crystal precursor.Spanish Ministry of Science through Juan de la Cierva-Formacion-2018 post-doctoral contract FJC2018-035820-IEuropean Union Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie Grant 101021894 (CARS-CO2) 844345 (EPIC)Polish National Agency for Academic Exchange (NAWA) PPI/APM/2018/1/00049/U/001Ministry of Science and Innovation, Spain (MICINN)Instituto de Salud Carlos IIISpanish Government PID2020-116660GB-I00European Cooperation in Science and Technology (COST) CA1713

Evolution of microstructure and residual stress during annealing of austenitic and ferritic steels

International audienceIn this work the recovery and recrystallization processes occurring in ferritic and austenitic steels were studied. To determine the evolution of residual stresses during material annealing the nonlinear sin 2 ψ diffraction method was used and an important relaxation of the macrostresses as well as the microstresses was found in the cold rolled samples subjected to heat treatment. Such relaxation occurs at the beginning of recovery, when any changes of microstructure cannot be detected using other experimental techniques. Stress evolution in the annealed steel samples was correlated with the progress of recovery process, which significantly depends on the value of stacking fault energy

Evolution of microstructure and residual stress during annealing of austenitic and ferritic steels

In this work the recovery and recrystallization processes occurring in ferritic and austenitic steels were studied. To determine the evolution of residual stresses during material annealing the nonlinear sin 2 ψ diffraction method was used and an important relaxation of the macrostresses as well as the microstresses was found in the cold rolled samples subjected to heat treatment. Such relaxation occurs at the beginning of recovery, when any changes of microstructure cannot be detected using other experimental techniques. Stress evolution in the annealed steel samples was correlated with the progress of recovery process, which significantly depends on the value of stacking fault energy

Anisotropy of mechanical properties of pinctada margaritifera mollusk shell

The mechanical properties such as compressive strength and nanohardness were investigated for Pinctada margaritifera mollusk shells. The compressive strength was evaluated through a uniaxial static compression test performed along the load directions parallel and perpendicular to the shell axis, respectively, while the hardness and Young modulus were measured using nanoindentation. In order to observe the crack propagation, for the first time for such material, the in-situ X-ray microscopy (nano-XCT) imaging (together with 3D reconstruction based on the acquired images) during the indentation tests was performed. The results were compared with these obtained during the micro-indentation test done with the help of conventional Vickers indenter and subsequent scanning electron microscopy observations. The results revealed that the cracks formed during the indentation start to propagate in the calcite prism until they reach a ductile organic matrix where most of them are stopped. The obtained results confirm a strong anisotropy of both crack propagation and the mechanical strength caused by the formation of the prismatic structure in the outer layer of P. margaritifera shell.The research was co-financed by the European Union from the resources of the European Social Fund (Project No.WND-POWR.03.02.00-00-I043/16), the DAAD program. A.C. acknowledges project CGL2017-85118-P of the Spanish Ministerio de Ciencia e Innovación for funding

Crystallographic control on the substructure of nacre tablets

Nacre tablets of mollusks develop two kinds of features when either the calcium carbonate or the organic portions are removed: (1) parallel lineations (vermiculations) formed by elongated carbonate rods, and (2) hourglass patterns, which appear in high relief when etched or in low relief if bleached. In untreated tablets, SEM and AFM data show that vermiculations correspond to aligned and fused aragonite nanogloblules, which are partly surrounded by thin organic pellicles. EBSD mapping of the surfaces of tablets indicates that the vermiculations are invariably parallel to the crystallographic a-axis of aragonite and that the triangles are aligned with the b-axis and correspond to the advance of the {010} faces during the growth of the tablet. According to our interpretation, the vermiculations appear because organic molecules during growth are expelled from the a-axis, where the Ca–CO3 bonds are the shortest. In this way, the subunits forming nacre merge uninterruptedly, forming chains parallel to the a-axis, whereas the organic molecules are expelled to the sides of these chains. Hourglass patterns would be produced by preferential adsorption of organic molecules along the {010}, as compared to the {100} faces. A model is presented for the nanostructure of nacre tablets. SEM and EBSD data also show the existence within the tablets of nanocrystalline units, which are twinned on {110} with the rest of the tablet. Our study shows that the growth dynamics of nacre tablets (and bioaragonite in general) results from the interaction at two different and mutually related levels: tablets and nanogranules

Crystallographic orientation inhomogeneity and crystal splitting in biogenic calcite

The calcitic prismatic units forming the outer shell of the bivalve Pinctada margaritifera have been analysed using scanning electron microscopy–electron back-scatter diffraction, transmission electron microscopy and atomic force microscopy. In the initial stages of growth, the individual prismatic unit sare single crystals. Their crystalline orientation is not consistent but rather changes gradually during growth. The gradients in crystallographic orientation occurmainly in a direction parallel to the long axis of the prism, i.e. perpendicular to the shell surface anddo not showpreferential tilting along anyof the calcite lattice axes. At a certain growth stage, gradients begin to spread and diverge, implying that the prismatic units split into several crystalline domains. In this way, a branched crystal, inwhich the ends of the branches are independent crystalline domains, is formed. At the nanometre scale, the material is composed of slightly misoriented domains, which are separated by planes approximately perpendicular to the c-axis. Orientational gradients and splitting processes are described in biocrystals for the first time and are undoubtedly related to the high content of intracrystalline organic molecules, although the way in which these act to induce the observed crystalline patterns is amatter of future research

Warunkowe zawieszenie wykonania kary w założeniach nowej polityki karnej

Opracowanie stanowi interdyscyplinarne studium badań nad instytucją warunkowego zawieszenia wykonania kary z perspektywy analizy dogmatycznej w płaszczyźnie prawa karnego materialnego i wynikających z niego implikacji w sferze proceduralnej oraz szeroko pojętej praktyki stosowania prawa karnego: sędziowskiej, prokuratorskiej i adwokacko-radcowskiej. Wiele miejsca poświęcono zagadnieniom z zakresu teorii i filozofii prawa karnego, umiejętnie eksponując przy tym ich wymiar praktyczny. Omówiono m.in. ustawowe przesłanki stosowania warunkowego zawieszenia, jego wady i zalety, obowiązki probacyjne związane z poddaniem sprawcy próbie