TY - JOUR A1 - Reinmuth-Selzle, K. A1 - Tchipilov, Teodor A1 - Backes, A. T. A1 - Tscheuschner, Georg A1 - Tang, K. A1 - Ziegler, K. A1 - Lucas, K. A1 - Pöschl, U. A1 - Fröhlich-Nowoisky, J. A1 - Weller, Michael G. T1 - Determination of the protein content of complex samples by aromatic amino acid analysis, liquid chromatography-UV absorbance, and colorimetry N2 - Fast and accurate determination of the protein content of a sample is an important and non-trivial task of many biochemical, biomedical, food chemical, pharmaceutical, and environmental research activities. Different methods of total protein determination are used for a wide range of proteins with highly variable properties in complex matrices. These methods usually work reasonably well for proteins under controlled conditions, but the results for non-standard and complex samples are often questionable. Here, we compare new and well-established methods, including traditional amino acid analysis (AAA), aromatic amino acid analysis (AAAA) based on the amino acids phenylalanine and tyrosine, reversed-phase liquid chromatography of intact proteins with UV absorbance measurements at 220 and 280 nm (LC-220, LC-280), and colorimetric assays like Coomassie Blue G-250 dye-binding assay (Bradford) and bicinchoninic acid (BCA) assay. We investigated different samples, including proteins with challenging properties, chemical modifications, mixtures, and complex matrices like air particulate matter and pollen extracts. All methods yielded accurate and precise results for the protein and matrix used for calibration. AAA, AAAA with fluorescence detection, and the LC-220 method yielded robust results even under more challenging conditions (variable analytes and matrices). These methods turned out to be well-suited for reliable determination of the protein content in a wide range of samples, such as air particulate matter and pollen. KW - Air particulate matter KW - Aromatic amino acid analysis KW - Atmospheric aerosol KW - Chemical protein modification KW - Derivatization KW - Nitration KW - Nitrotyrosine KW - LC-UV absorbance KW - Pollen extract KW - Protein quantification KW - Protein test KW - Kjeldahl KW - Tyrosine KW - Phenylalanine KW - Hydrolysis KW - Bradford KW - BCA test KW - 280 nm KW - Air filter samples KW - Fluorescence KW - HPLC KW - Chromatography KW - Protein content KW - 150th anniversary of BAM KW - Topical collection: Analytical Methods and Applications in the Materials and Life Sciences PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-545924 UR - https://pubmed.ncbi.nlm.nih.gov/35320366/ DO - https://doi.org/10.1007/s00216-022-03910-1 SP - 1 EP - 14 PB - Springer Nature Limited CY - New York, Heidelberg AN - OPUS4-54592 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Reinmuth-Selzle, K. A1 - Kampf, C. J. A1 - Lucas, K. A1 - Lang-Yona, N. A1 - Fröhlich-Nowoisky, J. A1 - Shiraiwa, M. A1 - Lakey, P. S. J. A1 - Lai, S. A1 - Liu, F. A1 - Kunert, A. T. A1 - Ziegler, K. A1 - Shen, F. A1 - Sgarbanti, R. A1 - Weber, B. A1 - Bellinghausen, I. A1 - Saloga, J. A1 - Weller, Michael G. A1 - Duschl, A. A1 - Schuppan, D. A1 - Pöschl, U. T1 - Air pollution and climate change effects on allergies in the anthropocene: Abundance, interaction, and modification of allergens and adjuvants N2 - Air pollution and climate change are potential drivers for the increasing burden of allergic diseases. The molecular mechanisms by which air pollutants and climate parameters may influence allergic diseases, however, are complex and elusive. This article provides an overview of physical, chemical and biological interactions between air pollution, climate change, allergens, adjuvants and the immune system, addressing how these interactions may promote the development of allergies. We reviewed and synthesized key findings from atmospheric, climate, and biomedical research. The current state of knowledge, open questions, and future research perspectives are outlined and discussed. The Anthropocene, as the present era of globally pervasive anthropogenic influence on planet Earth and, thus, on the human environment, is characterized by a strong increase of carbon dioxide, ozone, nitrogen oxides, and combustion- or traffic-related particulate matter in the atmosphere. These environmental factors can enhance the abundance and induce chemical modifications of allergens, increase oxidative stress in the human body, and skew the immune system toward allergic reactions. In particular, air pollutants can act as adjuvants and alter the immunogenicity of allergenic proteins, while climate change affects the atmospheric abundance and human exposure to bioaerosols and aeroallergens. To fully understand and effectively mitigate the adverse effects of air pollution and climate change on allergic diseases, several challenges remain to be resolved. Among these are the identification and quantification of immunochemical reaction pathways involving allergens and adjuvants under relevant environmental and physiological conditions. KW - Allergie KW - Klimaveränderung KW - Luftverschmutzung KW - Partikel KW - Ozon KW - Stickoxide KW - Allergene KW - Adjuvantien KW - PALM KW - DAMP PY - 2017 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-404531 DO - https://doi.org/10.1021/acs.est.6b04908 SN - 1520-5851 SN - 0013-936X VL - 51 IS - 8 SP - 4119 EP - 4141 PB - American Chemical Society (ACS) CY - Washington AN - OPUS4-40453 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Reinmuth-Selzle, K. A1 - Bellinghausen, I. A1 - Leifke, A. L. A1 - Backes, A. T. A1 - Bothen, N. A1 - Ziegler, K. A1 - Weller, Michael G. A1 - Saloga, J. A1 - Schuppan, D. A1 - Lucas, K. A1 - Pöschl, U. A1 - Fröhlich-Nowoisky, J. T1 - Chemical modification by peroxynitrite enhances TLR4 activation of the grass pollen allergen Phl p 5 N2 - The chemical modification of aeroallergens by reactive oxygen and nitrogen species (ROS/RNS) may contribute to the growing prevalence of respiratory allergies in industrialized countries. Post-translational modifications can alter the immunological properties of proteins, but the underlying mechanisms and effects are not well understood. In this study, we investigate the Toll-like receptor 4 (TLR4) activation of the major birch and grass pollen allergens Bet v 1 and Phl p 5, and how the physiological oxidant peroxynitrite (ONOO–) changes the TLR4 activation through protein nitration and the formation of protein dimers and higher oligomers. Of the two allergens, Bet v 1 exhibited no TLR4 activation, but we found TLR4 activation of Phl p 5, which increased after modification with ONOO– and may play a role in the sensitization against this grass pollen allergen. We attribute the TLR4 activation mainly to the two-domain structure of Phl p 5 which may promote TLR4 dimerization and activation. The enhanced TLR4 signaling of the modified allergen indicates that the ONOO–-induced modifications affect relevant protein-receptor interactions. This may lead to increased sensitization to the grass pollen allergen and thus contribute to the increasing prevalence of allergies in the Anthropocene, the present era of globally pervasive anthropogenic influence on the environment. KW - Bet v 1 KW - Birch pollen allergen KW - Phl p 5 KW - Grass pollen KW - Phleum pratense KW - Betula pendula KW - Nitration KW - Nitrotyrosine KW - Protein nitration KW - Toll-like receptor 4 KW - Allergy KW - Enhancement KW - Oligomerization KW - Dimerization KW - TLR4 activation KW - Air pollution KW - Nitrogen oxides KW - Inflammation PY - 2023 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-570298 DO - https://doi.org/10.3389/falgy.2023.1066392 VL - 4 SP - 1 EP - 7 PB - Frontiers Media SA CY - Lausanne, Switzerland AN - OPUS4-57029 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Ziegler, K. A1 - Kunert, A. T. A1 - Reinmuth-Selzle, K. A1 - Leifke, A. L. A1 - Widera, D. A1 - Weller, Michael G. A1 - Schuppan, D. A1 - Fröhlich-Nowoisky, J. A1 - Lucas, K. A1 - Pöschl, U. T1 - Chemical modification of pro-inflammatory proteins by peroxynitrite increases activation of TLR4 and NF-κB: Implications for the health effects of air pollution and oxidative stress N2 - Environmental pollutants like fine particulate matter can cause adverse health effects through oxidative stress and inflammation. Reactive oxygen and nitrogen species (ROS/RNS) such as peroxynitrite can chemically modify proteins, but the effects of such modifications on the immune system and human health are not well understood. In the course of inflammatory processes, the Toll-like receptor 4 (TLR4) can sense damage-associated molecular patterns (DAMPs). Here, we investigate how the TLR4 response and pro-inflammatory potential of the proteinous DAMPs α-Synuclein (α-Syn), heat shock protein 60 (HSP60), and high-mobility-group box 1 protein (HMGB1), which are relevant in neurodegenerative and cardiovascular diseases, changes upon chemical modification with peroxynitrite. For the peroxynitrite-modified proteins, we found a strongly enhanced activation of TLR4 and the pro-inflammatory transcription factor NF-κB in stable reporter cell lines as well as increased mRNA expression and secretion of the pro-inflammatory cytokines TNF-α, IL-1β, and IL-8 in human monocytes (THP-1). This enhanced activation of innate immunity via TLR4 is mediated by covalent chemical modifications of the studied DAMPs. Our results show that proteinous DAMPs modified by peroxynitrite more potently amplify inflammation via TLR4 activation than the native DAMPs, and provide first evidence that such modifications can directly enhance innate immune responses via a defined receptor. These findings suggest that environmental pollutants and related ROS/RNS may play a role in promoting acute and chronic inflammatory disorders by structurally modifying the body's own DAMPs. This may have important consequences for chronic neurodegenerative, cardiovascular or gastrointestinal diseases that are prevalent in modern societies, and calls for action, to improve air quality and climate in the Anthropocene. KW - Protein nitration KW - Protein oligomerization KW - Damage-associated molecular patterns (DAMPs) KW - Pattern recognition receptor KW - Anthropocene KW - Environmental pollutants PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-517128 DO - https://doi.org/10.1016/j.redox.2020.101581 VL - 37 SP - 101581 PB - Elsevier B.V. AN - OPUS4-51712 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Backes, A. T. A1 - Reinmuth-Selzle, K. A1 - Leifke, A. L. A1 - Ziegler, K. A1 - Krevert, C. S. A1 - Tscheuschner, Georg A1 - Lucas, K. A1 - Weller, Michael G. A1 - Berkemeier, T. A1 - Pöschl, U. A1 - Fröhlich-Nowoisky, J. T1 - Oligomerization and Nitration of the Grass Pollen Allergen Phl p 5 by Ozone, Nitrogen Dioxide, and Peroxynitrite: Reaction Products, Kinetics, and Health Effects N2 - The allergenic and inflammatory potential of proteins can be enhanced by chemical modification upon exposure to atmospheric or physiological oxidants. The molecular mechanisms and kinetics of such modifications, however, have not yet been fully resolved. We investigated the oligomerization and nitration of the grass pollen allergen Phl p 5 by ozone (O3), nitrogen dioxide (NO2), and peroxynitrite (ONOO–). Within several hours of exposure to atmospherically relevant concentration levels of O3 and NO2, up to 50% of Phl p 5 were converted into protein oligomers, likely by formation of dityrosine cross-links. Assuming that tyrosine residues are the preferential site of nitration, up to 10% of the 12 tyrosine residues per protein monomer were nitrated. For the reaction with peroxynitrite, the largest oligomer mass fractions (up to 50%) were found for equimolar concentrations of peroxynitrite over tyrosine residues. With excess peroxynitrite, the nitration degrees increased up to 40% whereas the oligomer mass fractions decreased to 20%. Our results suggest that protein oligomerization and nitration are competing processes, which is consistent with a two-step mechanism involving a reactive oxygen intermediate (ROI), as observed for other proteins. The modified proteins can promote pro-inflammatory cellular signaling that may contribute to chronic inflammation and allergies in response to air pollution. KW - Protein KW - Nitrotyrosine KW - Dityrosine KW - Allergy KW - Hay fever KW - Diesel exhaust KW - Combustion KW - Exhaust PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-529729 DO - https://doi.org/10.3390/ijms22147616 VL - 22 IS - 14 PB - MDPI CY - Basel AN - OPUS4-52972 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Puthiyaveettil, N. A1 - Thomas, K. R. A1 - Myrach, Philipp A1 - Ziegler, Mathias A1 - Rajagopal, P. A1 - Balasubramaniamam, K. T1 - Defect detection in steel bars up to 600 °C using laser line thermography N2 - Crack detection in steel bars at high surface temperatures is a critical problem in any manufacturing industry. Surface breaking cracks are the major problems during the billet casting. Many NDT techniques are proven its capability in crack detection at room temperature. Here, we are demonstrating the possibility of exposure of cracks using laser line thermography at higher surface temperatures (up to 600 °C). A continuous-wave (CW) laser is used to excite the sample kept at higher surface temperatures. The temperature distribution over the sample due to the laser line scanning is captured using a temperature calibrated infrared (IR) thermal camera. The response of the sample temperature in crack detection is investigated using a validated FE model. The impact of the oxide layer in crack detection is investigated by using two types of samples; one without any oxide layer and the second is with the oxide layer. The influence of laser power in the detection of defects at high temperatures is studied. 3D numerical models were developed for the cases; when the sample is with oxide layer and without any oxide layer for a better understanding of physics. The surface temperature rise due to laser heating is higher for the scaled sample compared to the no-scale sample. The presence of the oxide layer above the parent metal will reduce the reflectivity of the surface. Lower reflectivity will lead to increased absorption of incident energy so that the surface temperature rise will be higher than the surface with no scale. Thermal contrast linearly depends on laser power, which means higher laser power will increase the defect detectability even at a higher surface temperature. KW - Laser thermography KW - High temperature KW - Modeling KW - Surface cracks KW - Non-destructive testing PY - 2020 DO - https://doi.org/10.1016/j.infrared.2020.103565 SN - 1350-4495 VL - 111 SP - 103565 PB - Elsevier B.V. AN - OPUS4-51573 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Puthiyaveettil, N. A1 - Thomas K, R. A1 - Unnikrishnakurup, S. A1 - Myrach, Philipp A1 - Ziegler, Mathias A1 - Balasubramaniam, K. T1 - Laser line scanning thermography for surface breaking crack detection modeling and experimental study N2 - Crack detection in metallic samples at high surface temperature, hostile and hazardous environments, etc. is challenging situation in any manufacturing industries. Most of the present NDE methods are suitable only for lower surface temperatures, especially room temperature. In this situation, we need a fast and non-contact NDT method which can be applied even in high sample surface temperature. Laser thermography is one of the techniques having a high potential in non-contact inspection. As a preliminary investigation, in this article, we have studied the potentiality of laser line thermography in crack detection at room temperature. In laser line thermography, a continuous wave (CW) laser is used to generate a laser line, which in turn is used to scan the metal surface. The heat distribution over the sample surface is recorded by an infrared thermal (IR) camera. Two different approaches are reported in this work. Firstly, a stationary laser line source and its interaction with cracks; secondly, moving laser line source scanning over a surface with crack. When the distance between crack centre to laser line centre increases, crack detectability will decrease; and when laser power increases, crack detectability will increase. A dedicated image processing algorithm was developed to improve the detectability of the cracks. To understand the heat transfer phenomenon, a simplified 3D model for laser thermography was developed for the heat distribution during laser heating and was validated with experimental results. Defects were incorporated as a thermally thin resistive layer (TTRL) in numerical modeling, and the effect of TTRL in heat conduction is compared with experimental results. KW - Thermography KW - Laser Thermography KW - Cracks KW - FEM KW - NDT PY - 2019 DO - https://doi.org/10.1016/j.infrared.2019.103141 VL - 104 SP - 103141 PB - Elsevier B.V. AN - OPUS4-49941 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Puthiyaveettil, N. A1 - Kidangan, R. A1 - Unnikrishnakurup, Sreedhar A1 - Krishnamurthy, C. V. A1 - Ziegler, Mathias A1 - Myrach, Philipp A1 - Balasubramaniam, K. T1 - Numerical study of laser line thermography for crack detection at high temperature N2 - The detection of cracks before the failure is highly significant when it comes to safety-relevant structures. Crack detection in metallic samples at high surface temperature is one of the challenging situation in manufacturing industries. Laser thermography has already proved its detection capability of surface cracks in metallic samples at room temperature. In this work a continuous wave (CW) laser use to generate a laser, which is using to scan the metal surface with notch. The corresponding heat distribution on the surface monitored using infrared thermal (IR) camera. A simplified 3D model for laser thermography is developed and validated with experimental results. A dedicated image processing algorithm developed to improve the detectability of the cracks. To understand the dependency of surface temperature, laser power, laser scanning speed etc. in defect detection, we carried out parametric studies with our validated model. Here we Report the capability of laser thermography in crack detection at elevated temperature. T2 - 14th Quantitative InfraRed Thermography Conference CY - Berlin, Germany DA - 25.06.2018 KW - Thermal contrast KW - Laser Thermography KW - FEM KW - Surface cracks KW - NDT PY - 2018 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-499113 DO - https://doi.org/10.21611/qirt.2018.076 SN - 2371-4085 VL - 2018 SP - 685 EP - 686 AN - OPUS4-49911 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schweikhard, E. S. A1 - Kuhlmann, S. I. A1 - Kunte, Hans-Jörg A1 - Grammann, K. A1 - Ziegler, C. M. T1 - Structure and function of the universal stress protein TeaD and its role in regulating the ectoine transporter TeaABC of halomonas elongata DSM 2581T N2 - The halophilic bacterium Halomonas elongata takes up the compatible solute ectoine via the osmoregulated TRAP transporter TeaABC. A fourth orf (teaD) is located adjacent to the teaABC locus that encodes a putative universal stress protein (USP). By RT-PCR experiments we proved a cotranscription of teaD along with teaABC. Deletion of teaD resulted in an enhanced uptake for ectoine by the Transporter TeaABC and hence a negative activity regulation of TeaABC by TeaD. Atranscriptional regulation viaDNA binding could be excluded. ATP binding to native TeaD was shown by HPLC, and the Crystal structure of TeaD was solved in complex with ATP to a resolution of 1.9 A ° by molecular replacement. TeaD forms a dimer-dimer complex with one ATP molecule bound to each monomer, which has a Rossmann-like R/β overall fold. Our results reveal an ATP-dependent oligomerization of TeaD, which might have a functional role in the regulatory mechanism of TeaD. USP-encoding orfs, which are located adjacent to genes Encoding for TeaABC homologues, could be identified in several other organisms, and their physiological role in balancing the internal cellular ectoine pool is discussed. KW - Universal stress protein KW - Ectoine KW - Transporter TeaABC PY - 2010 DO - https://doi.org/10.1021/bi9017522 SN - 1520-4995 SN - 0006-2960 VL - 49 IS - 10 SP - 2194 EP - 2204 AN - OPUS4-37682 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Krishna, S. K. P. A1 - Puthiyaveetil, N. A1 - Kidangan, R. A1 - Unnikrishnakurup, Sreedhar A1 - Ziegler, Mathias A1 - Myrach, Philipp A1 - Balasubramaniam, Krishnan A1 - Purushothaman, B. T1 - Raw data based image processing algorithm for fast detection of surface breaking cracks N2 - The aim of this work is to illustrate the contribution of signal processing techniques in the field of Non-Destructive Evaluation. A component’s life evaluation is inevitably related to the presence of flaws in it. The detection and characterization of cracks prior to damage is a technologically and economically significant task and is of very importance when it comes to safety-relevant measures. The Laser Thermography is the most effective and advanced thermography method for Non-Destructive Evaluation. High capability for the detection of surface cracks and for the characterization of the geometry of artificial surface flaws in metallic samples of laser thermography is particularly encouraging. This is one of the non- contacting, fast and real time detection method. The presence of a vertical surface breaking crack will disturb the thermal footprint. The data processing method plays vital role in fast detection of the surface and sub-surface cracks. Currently in laser thermographic inspection lacks a compromising data processing algorithm which is necessary for the fast crack detection and also the analysis of data is done as part of post processing. In this work we introduced a raw data based image processing algorithm which results precise, better and fast crack detection. The algorithm we developed gives better results in both experimental and modeling data. By applying this algorithm we carried out a detailed investigation Variation of thermal contrast with crack parameters like depth and width. The algorithm we developed is applied for various surface temperature data from the 2D scanning model and also validated credibility of algorithm with experimental data. T2 - QNDE conference 2016 - Review of progress in quantitative nondestructive evaluation CY - Atlanta, GA, USA DA - 18.07.2016 KW - Crack Detection KW - Laserthermografie KW - Thermografie KW - Risserkennung PY - 2017 SN - 978-0-7354-1474-7 DO - https://doi.org/10.1063/1.4974723 SN - 0094-243X VL - 1806 IS - 1 SP - UNSP 140008, 1 EP - 9 PB - AIP Publishing CY - New York AN - OPUS4-39387 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -