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Purpose: Human OX40 (hOX40/CD134), a member of the TNF receptor superfamily, is mainly expressed on activated T lymphocytes. Triggered by its ligand OX40L (CD252), it provides costimulatory signals that support the differentiation, proliferation and long-term survival of T cells. Besides being a relevant therapeutic target, hOX40 is also an important biomarker for monitoring the presence or infiltration of activated T cells within the tumor microenvironment (TME), the inflammatory microenvironment (IME) in immune-mediated diseases (IMIDs) and the lymphatic organs. Here, we developed novel single domain antibodies (nanobodies, Nbs) targeting hOX40 to monitor the activation status of T cells by in vivo molecular imaging., Methods: Nbs against hOX40 (hOX40-Nbs) were selected from an immunized Nb-library by phage display. The identified hOX40-Nbs were characterized in vitro , including determination of their specificity, affinity, stability, epitope recognition and their impact on OX40 signaling and T cell function. A lead candidate was site-specifically conjugated with a fluorophore via sortagging and applied for noninvasive in vivo optical imaging (OI) of hOX40-expressing cells in a xenograft mouse model., Results: Our selection campaign revealed four unique Nbs that exhibit strong binding affinities and high stabilities under physiological conditions. Epitope binning and domain mapping indicated the targeting of at least two different epitopes on hOX40. When analyzing their impact on OX40 signaling, an agonistic effect was excluded for all validated Nbs. Incubation of activated T cells with hOX40-Nbs did not affect cell viability or proliferation patterns, whereas differences in cytokine release were observed. In vivo OI with a fluorophore-conjugated lead candidate in experimental mice with hOX40-expressing xenografts demonstrated its specificity and functionality as an imaging probe., Conclusion: Considering the need for advanced probes for noninvasive in vivo monitoring of T cell activation dynamics, we propose, that our hOX40-Nbs have a great potential as imaging probes for noninvasive and longitudinal in vivo diagnostics. Quantification of OX40 + T cells in TME or IME will provide crucial insights into the activation state of infiltrating T cells, offering a valuable biomarker for assessing immune responses, predicting treatment efficacy, and guiding personalized immunotherapy strategies in patients with cancer or IMIDs., Competing Interests: DF, DS, MK, BP, TW, BT, PK, and UR are named as inventors on a patent application claiming the use of the described nanobodies for diagnosis and therapeutics filed by the Natural and Medical Sciences Institute, the Werner Siemens Imaging Center and the University of Tuebingen. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Frecot, Blaess, Wagner, Kaiser, Traenkle, Fandrich, Jakobi, Scholz, Nueske, Schneiderhan-Marra, Gouttefangeas, Kneilling, Pichler, Sonanini and Rothbauer.)

Background and Purpose: Parkinson's disease (PD) is an age-related condition characterized by substantial phenotypic variability. Consequently, pathways and proteins involved in biological aging, such as the central aging pathway comprising insulin-like growth factor 1-α-Klotho-sirtuin 1-forkhead box O3-peroxisome proliferator-activated receptor γ, may potentially influence disease progression., Methods: Cerebrospinal fluid (CSF) levels of α-Klotho in 471 PD patients were examined. Of the 471 patients, 96 carried a GBA1 variant (PD GBA1), whilst the 375 non-carriers were classified as PD wild-type (PD WT). Each patient was stratified into a CSF α-Klotho tertile group based on the individual level. Kaplan-Meier survival curves and Cox regression analysis stratified by tertile groups were conducted. These longitudinal data were available for 255 patients. Follow-up times reached from 8.4 to 12.4 years. The stratification into PD WT and PD GBA1 was undertaken to evaluate potential continuum patterns, particularly in relation to CSF levels., Results: Higher CSF levels of α-Klotho were associated with a significant later onset of cognitive impairment. Elevated levels of α-Klotho in CSF were linked to higher Montreal Cognitive Assessment scores in male PD patients with GBA1 mutations., Conclusions: Our results indicate that higher CSF levels of α-Klotho are associated with a delayed cognitive decline in PD. Notably, this correlation is more prominently observed in PD patients with GBA1 mutations, potentially reflecting the accelerated biological aging profile characteristic of individuals harboring GBA1 variants., (© 2024 The Author(s). European Journal of Neurology published by John Wiley & Sons Ltd on behalf of European Academy of Neurology.)

Background: Prior investigations have elucidated pathophysiological interactions involving blood coagulation and neurodegenerative diseases. These interactions pertain to age-related effects and a mild platelet antiaggregant function of exogenous α-Synuclein., Objective: Our study sought to explore whether cerebrospinal fluid (CSF) levels of tissue factor (TF), the initiator of the extrinsic pathway of hemostasis, differ between controls (CON) compared to patients with Parkinson's disease (PD) and dementia with Lewy bodies (DLB), considering that these conditions represent a spectrum of α-Synuclein pathology. We further investigated whether TF levels are associated with longitudinal progression in PD., Methods: We examined CSF levels of TF in 479 PD patients, 67 patients diagnosed with DLB, and 16 CON in order to evaluate potential continuum patterns among DLB, PD, and CON. Of the 479 PD patients, 96 carried a GBA1 variant (PD GBA1), while the 383 non-carriers were classified as PD wildtype (PD WT). We considered both longitudinal clinical data as well as CSF measurements of common neurodegenerative markers (amyloid-β 1-42, h-Tau, p-Tau, NfL, α-Synuclein). Kaplan-Meier survival and Cox regression analysis stratified by TF tertile levels was conducted., Results: Higher CSF levels of TF were associated with an older age at examination in PD and a significant later onset of postural instability in PD GBA1. TF levels were lower in male vs. female PD. DLB GBA1 exhibited the lowest TF levels, followed by PD GBA1, with CON showing the highest levels., Conclusions: TF as representative of blood hemostasis could be an interesting CSF candidate to further explore in PD and DLB.

Many modern automated vehicle sensor systems use light detection and ranging (LiDAR) sensors. The prevailing technology is scanning LiDAR, where a collimated laser beam illuminates objects sequentially point-by-point to capture 3D range data. In current systems, the point clouds from the LiDAR sensors are mainly used for object detection. To estimate the velocity of an object of interest (OoI) in the point cloud, the tracking of the object or sensor data fusion is needed. Scanning LiDAR sensors show the motion distortion effect, which occurs when objects have a relative velocity to the sensor. Often, this effect is filtered, by using sensor data fusion, to use an undistorted point cloud for object detection. In this study, we developed a method using an artificial neural network to estimate an object's velocity and direction of motion in the sensor's field of view (FoV) based on the motion distortion effect without any sensor data fusion. This network was trained and evaluated with a synthetic dataset featuring the motion distortion effect. With the method presented in this paper, one can estimate the velocity and direction of an OoI that moves independently from the sensor from a single point cloud using only one single sensor. The method achieves a root mean squared error (RMSE) of 0.1187 m s -1 and a two-sigma confidence interval of [-0.0008 m s -1 , 0.0017 m s -1 ] for the axis-wise estimation of an object's relative velocity, and an RMSE of 0.0815 m s -1 and a two-sigma confidence interval of [0.0138 m s -1 , 0.0170 m s -1 ] for the estimation of the resultant velocity. The extracted velocity information (4D-LiDAR) is available for motion prediction and object tracking and can lead to more reliable velocity data due to more redundancy for sensor data fusion.

In this work, we introduce a novel approach to model the rain and fog effect on the light detection and ranging (LiDAR) sensor performance for the simulation-based testing of LiDAR systems. The proposed methodology allows for the simulation of the rain and fog effect using the rigorous applications of the Mie scattering theory on the time domain for transient and point cloud levels for spatial analyses. The time domain analysis permits us to benchmark the virtual LiDAR signal attenuation and signal-to-noise ratio (SNR) caused by rain and fog droplets. In addition, the detection rate (DR), false detection rate (FDR), and distance error derror of the virtual LiDAR sensor due to rain and fog droplets are evaluated on the point cloud level. The mean absolute percentage error (MAPE) is used to quantify the simulation and real measurement results on the time domain and point cloud levels for the rain and fog droplets. The results of the simulation and real measurements match well on the time domain and point cloud levels if the simulated and real rain distributions are the same. The real and virtual LiDAR sensor performance degrades more under the influence of fog droplets than in rain.

Measurement performance evaluation of real and virtual automotive light detection and ranging (LiDAR) sensors is an active area of research. However, no commonly accepted automotive standards, metrics, or criteria exist to evaluate their measurement performance. ASTM International released the ASTM E3125-17 standard for the operational performance evaluation of 3D imaging systems commonly referred to as terrestrial laser scanners (TLS). This standard defines the specifications and static test procedures to evaluate the 3D imaging and point-to-point distance measurement performance of TLS. In this work, we have assessed the 3D imaging and point-to-point distance estimation performance of a commercial micro-electro-mechanical system (MEMS)-based automotive LiDAR sensor and its simulation model according to the test procedures defined in this standard. The static tests were performed in a laboratory environment. In addition, a subset of static tests was also performed at the proving ground in natural environmental conditions to determine the 3D imaging and point-to-point distance measurement performance of the real LiDAR sensor. In addition, real scenarios and environmental conditions were replicated in the virtual environment of a commercial software to verify the LiDAR model's working performance. The evaluation results show that the LiDAR sensor and its simulation model under analysis pass all the tests specified in the ASTM E3125-17 standard. This standard helps to understand whether sensor measurement errors are due to internal or external influences. We have also shown that the 3D imaging and point-to-point distance estimation performance of LiDAR sensors significantly impacts the working performance of the object recognition algorithm. That is why this standard can be beneficial in validating automotive real and virtual LiDAR sensors, at least in the early stage of development. Furthermore, the simulation and real measurements show good agreement on the point cloud and object recognition levels.

Inflammation modifies the incidence and progression of Parkinson's disease (PD). By using 30 inflammatory markers in CSF in 498 people with PD and 67 people with dementia with Lewy bodies (DLB) we show that: (1) levels of ICAM-1, Interleukin-8, MCP-1, MIP-1 beta, SCF and VEGF were associated with clinical scores and neurodegenerative CSF biomarkers (Aβ1-42, t-Tau, p181-Tau, NFL and α-synuclein). (2) PD patients with GBA mutations show similar levels of inflammatory markers compared to PD patients without GBA mutations, even when stratified by mutation severity. (3) PD patients who longitudinally developed cognitive impairment during the study had higher levels of TNF-alpha at baseline compared to patients without the development of cognitive impairment. (4) Higher levels of VEGF and MIP-1 beta were associated with a longer duration until the development of cognitive impairment. We conclude that the majority of inflammatory markers is limited in robustly predicting longitudinal trajectories of developing cognitive impairment., (© 2023. The Author(s).)

Autonomous vehicles need accurate 3D perception with a decent frame rate and high angular resolution to detect obstacles reliably and avoid collisions. We developed a low-cost scanning multichannel light detection and ranging sensor architecture allowing scalable frame rates by adjusting the number of laser and detector pairs. Scanning is achieved by a pair of micro-electro-mechanical system (MEMS) mirrors. A control pattern for the MEMS mirrors to maximize the frame rate is presented. A built prototype based on the proposed architecture achieves a frame rate of 11.5 Hz, a field of view of 70 ∘ ×30 ∘ , and an angular resolution of 0.4°. The distance resolution is 6 cm. Reliable single-shot detection for low-reflective objects up to 19 m indoors and 11 m under direct sunlight exposure is achieved. A performance assessment based on the presented measurement system for recently available vertical-cavity surface-emitting laser arrays with power densities up to 1 k W / m m 2 shows promising improvement potential.

Accurate and precise point-of-care (POC) testing for C-reactive protein (CRP) can help support healthcare providers in the clinical management of patients. Here, we compared the analytical performance of 17 commercially available POC CRP tests to enable more decentralized use of the tool. The following CRP tests were evaluated. Eight quantitative tests: QuikRead go (Aidian), INCLIX (Sugentech), Spinit (Biosurfit), LS4000 (Lansionbio), GS 1200 (Gensure Biotech), Standard F200 (SD Biosensor), Epithod 616 (DxGen), IFP-3000 (Xincheng Biological); and nine semi-quantitative tests: Actim CRP (ACTIM), NADAL Dipstick (nal von minden), NADAL cassette (nal von minden), ALLTEST Dipstick (Hangzhou Alltest Biotech), ALLTEST Cassette cut-off 10-40-80 (Hangzhou Alltest Biotech), ALLTEST Cassette cut-off 10-30 (Hangzhou Alltest Biotech), Biotest (Hangzhou Biotest Biotech), BTNX Quad Line (BTNX), BTNX Tri Line (BTNX). Stored samples (n = 660) had previously been tested for CRP using Cobas 8000 Modular analyzer (Roche Diagnostics International AG, Rotkreuz, Switzerland (reference standards). CRP values represented the clinically relevant range (10-100 mg/L) and were grouped into four categories (<10 mg/L, 10-40 mg/L or 10-30 mg/L, 40-80 mg/L or 30-80 mg/L, and > 80mg/L) for majority of the semi-quantitative tests. Among the eight quantitative POC tests evaluated, QuikRead go and Spinit exhibited better agreement with the reference method, showing slopes of 0.963 and 0.921, respectively. Semi-quantitative tests with the four categories showed a poor percentage agreement for the intermediate categories and higher percentage agreement for the lower and upper limit categories. Analytical performance varied considerably for the semi-quantitative tests, especially among the different categories of CRP values. Our findings suggest that quantitative tests might represent the best choice for a variety of use cases, as they can be used across a broad range of CRP categories., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Calarco et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This work introduces a process to develop a tool-independent, high-fidelity, ray tracing-based light detection and ranging (LiDAR) model. This virtual LiDAR sensor includes accurate modeling of the scan pattern and a complete signal processing toolchain of a LiDAR sensor. It is developed as a functional mock-up unit (FMU) by using the standardized open simulation interface (OSI) 3.0.2, and functional mock-up interface (FMI) 2.0. Subsequently, it was integrated into two commercial software virtual environment frameworks to demonstrate its exchangeability. Furthermore, the accuracy of the LiDAR sensor model is validated by comparing the simulation and real measurement data on the time domain and on the point cloud level. The validation results show that the mean absolute percentage error (MAPE) of simulated and measured time domain signal amplitude is 1.7%. In addition, the MAPE of the number of points Npoints and mean intensity Imean values received from the virtual and real targets are 8.5% and 9.3%, respectively. To the author's knowledge, these are the smallest errors reported for the number of received points Npoints and mean intensity Imean values up until now. Moreover, the distance error derror is below the range accuracy of the actual LiDAR sensor, which is 2 cm for this use case. In addition, the proving ground measurement results are compared with the state-of-the-art LiDAR model provided by commercial software and the proposed LiDAR model to measure the presented model fidelity. The results show that the complete signal processing steps and imperfections of real LiDAR sensors need to be considered in the virtual LiDAR to obtain simulation results close to the actual sensor. Such considerable imperfections are optical losses, inherent detector effects, effects generated by the electrical amplification, and noise produced by the sunlight.

An optical fiber with both temperature and strain fiber Bragg grating sensors were embedded into an aluminum cast structure during the casting process. Temperature and strain calibrations were carried out respectively for the metal-embedded sensors. Temperature and external strain decoupling was further demonstrated in a temperature range from 25 to 80 °C and an external strain range from 0 to ∼110 µɛ. With the interpolated temperature measured by two temperature sensors at different positions, the external strain could be decoupled from temperature and thermal strain at the strain sensor. The temperature and external strain values obtained from our embedded optical fiber sensors agreed well with reference values, revealing the good performance of the metal-embedded optical fiber sensors. The difference between the measured values and the reference values are within ±5 µɛ for external strain and ±1 °C for temperature. With only a single fiber, the in-situ temperature and external strain information in the aluminum structure can be monitored in real time, representing an important step towards fiber-optic smart casts. Our investigation demonstrates that embedded optical fiber sensors can be a promising method for structural health monitoring of metallic structures.

Background: An involvement of the central-nervous and peripheral, innate and adaptive immune system in the pathogenesis of Parkinson's disease (PD) is nowadays well established., Objectives: We face several open questions in preparation of clinical trials aiming at disease-modification by targeting the immune system: Do peripheral (blood) inflammatory profiles reflect central (CSF) inflammatory processes? Are blood/CSF inflammatory markers associated with CSF levels of neurodegenerative/PD-specific biomarkers?, Methods: Using a multiplex assay we assessed 41 inflammatory markers in CSF/serum pairs in 453 sporadic PD patients. We analyzed CSF/serum correlation as well as associations of inflammatory markers with clinical outcome measures (UPDRS-III, H&Y, MoCA) and with CSF levels of α-synuclein, Aβ 1-42 , t- Tau, p181-Tau and NFL. All analyses were stratified by sex as the immune system shows relevant sex-specific differences., Results: Correlations between CSF and serum were sparse and detected in only 25% (9 out of 36) of the analysable inflammatory markers in male PD patients and in only 38% (12 out of 32) of female PD patients. The most important pro-inflammatory mediators associated with motor and cognitive decline as well as with neurodegenerative/PD-specific biomarkers were FABP, ICAM-1, IL-8, MCP-1, MIP-1-beta, and SCF. Results were more robust for CSF than for serum., Interpretation: Levels of central-nervous and peripheral inflammatory markers might be regulated independently of each other with CSF inflammatory markers reflecting CNS pathology more accurately than peripheral markers. These findings along with sex-specific characteristics have to be considered when designing clinical trials aiming at disease-modification by targeting the immune system., Competing Interests: KW and IL are employed by Roboscreen GmbH which manufactures the ELISA kit for measurements of total human α-synuclein used in the present study. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Lerche, Zimmermann, Wurster, Roeben, Fries, Deuschle, Waniek, Lachmann, Gasser, Jakobi, Joos, Schneiderhan-Marra and Brockmann.)

Background: Exposure to phthalates during pregnancy may alter DNA methylation in the placenta, a crucial organ for the growth and development of the fetus., Objectives: We studied associations between urinary concentrations of phthalate biomarkers during pregnancy and placental DNA methylation., Methods: We measured concentrations of 11 phthalate metabolites in maternal spot urine samples collected between 22 and 29 gestational weeks in 202 pregnant women. We analyzed DNA methylation levels in placental tissue (fetal side) collected at delivery. We first investigated changes in global DNA methylation of repetitive elements Alu and LINE-1. We then performed an adjusted epigenome-wide association study using IlluminaHM450 BeadChips and identified differentially methylated regions (DMRs) associated with phthalate exposure., Results: Monobenzyl phthalate concentration was inversely associated with placental methylation of Alu repeats. Moreover, all phthalate biomarkers except for monocarboxy-iso-octyl phthalate and mono(2-ethyl-5-hydroxyhexyl) phthalate were associated with at least one DMR. All but three DMRs showed increased DNA methylation with increased phthalate exposure. The largest identified DMR (22 CpGs) was positively associated with monocarboxy-iso-nonyl phthalate and encompassed heat shock proteins (HSPA1A, HSPA1L). The remaining DMRs encompassed transcription factors and nucleotide exchange factors, among other genes., Conclusions: This is the first description of genome-wide modifications of placental DNA methylation in association with pregnancy exposure to phthalates. Our results suggest epigenetic mechanisms by which exposure to these compounds could affect fetal development. Of interest, four identified DMRs had been previously associated with maternal smoking, which may suggest particular sensitivity of these genomic regions to the effect of environmental contaminants., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Biomaterial characteristics such as surface topographies have been shown to modulate macrophage phenotypes. The standard methodologies to measure macrophage response to biomaterials are marker-based and invasive. Raman microspectroscopy (RM) is a marker-independent, noninvasive technology that allows the analysis of living cells without the need for staining or processing. In the present study, we analyzed human monocyte-derived macrophages (MDMs) using RM, revealing that macrophage activation by lipopolysaccharides (LPS), interferons (IFN), or cytokines can be identified by lipid composition, which significantly differs in M0 (resting), M1 (IFN-γ/LPS), M2a (IL-4/IL-13), and M2c (IL-10) MDMs. To identify the impact of a biomaterial on MDM phenotype and polarization, we cultured macrophages on titanium disks with varying surface topographies and analyzed the adherent MDMs with RM. We detected surface topography-induced changes in MDM biochemistry and lipid composition that were not shown by less sensitive standard methods such as cytokine expression or surface antigen analysis. Our data suggest that RM may enable a more precise classification of macrophage activation and biomaterial-macrophage interaction., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

LiDAR sensors are a key technology for enabling safe autonomous cars. For highway applications, such systems must have a long range, and the covered field of view (FoV) of >45° must be scanned with resolutions higher than 0.1°. These specifications can be met by modern MEMS scanners, which are chosen for their robustness and scalability. For the automotive market, these sensors, and especially the scanners within, must be tested to the highest standards. We propose a novel measurement setup for characterizing and validating these kinds of scanners based on a position-sensitive detector (PSD) by imaging a deflected laser beam from a diffuser screen onto the PSD. A so-called ray trace shifting technique (RTST) was used to minimize manual calibration effort, to reduce external mounting errors, and to enable dynamical one-shot measurements of the scanner's steering angle over large FoVs. This paper describes the overall setup and the calibration method according to a standard camera calibration. We further show the setup's capabilities by validating it with a statically set rotating stage and a dynamically oscillating MEMS scanner. The setup was found to be capable of measuring LiDAR MEMS scanners with a maximum FoV of 47° dynamically, with an uncertainty of less than 1%.

The advancement of new immunotherapies necessitates appropriate probes to monitor the presence and distribution of distinct immune cell populations. Considering the key role of CD4 + cells in regulating immunological processes, we generated novel single-domain antibodies [nanobodies (Nbs)] that specifically recognize human CD4. After in-depth analysis of their binding properties, recognized epitopes, and effects on T-cell proliferation, activation, and cytokine release, we selected CD4-specific Nbs that did not interfere with crucial T-cell processes in vitro and converted them into immune tracers for noninvasive molecular imaging. By optical imaging, we demonstrated the ability of a high-affinity CD4-Nb to specifically visualize CD4 + cells in vivo using a xenograft model. Furthermore, quantitative high-resolution immune positron emission tomography (immunoPET)/MR of a human CD4 knock-in mouse model showed rapid accumulation of 64 Cu-radiolabeled CD4-Nb1 in CD4 + T cell-rich tissues. We propose that the CD4-Nbs presented here could serve as versatile probes for stratifying patients and monitoring individual immune responses during personalized immunotherapy in both cancer and inflammatory diseases., Competing Interests: DSo, MK, BP, BT, PK, and UR are named as inventors on a patent application claiming the use of the described nanobodies for diagnosis and therapeutics filed by the Natural and Medical Sciences Institute and the Werner Siemens Imaging Center. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Traenkle, Kaiser, Pezzana, Richardson, Gramlich, Wagner, Seyfried, Weldle, Holz, Parfyonova, Nueske, Scholz, Zeck, Jakobi, Schneiderhan-Marra, Schaller, Maurer, Gouttefangeas, Kneilling, Pichler, Sonanini and Rothbauer.)

In utero exposure to environmental chemicals, such as synthetic phenols, may alter DNA methylation in different tissues, including placenta - a critical organ for fetal development. We studied associations between prenatal urinary biomarker concentrations of synthetic phenols and placental DNA methylation. Our study involved 202 mother-son pairs from the French EDEN cohort. Nine phenols were measured in spot urine samples collected between 22 and 29 gestational weeks. We performed DNA methylation analysis of the fetal side of placental tissues using the IlluminaHM450 BeadChips. We evaluated methylation changes of individual CpGs in an adjusted epigenome-wide association study (EWAS) and identified differentially methylated regions (DMRs). We performed mediation analysis to test whether placental tissue heterogeneity mediated the association between urinary phenol concentrations and DNA methylation. We identified 46 significant DMRs (≥5 CpGs) associated with triclosan (37 DMRs), 2,4-dichlorophenol (3), benzophenone-3 (3), methyl- (2) and propylparaben (1). All but 2 DMRs were positively associated with phenol concentrations. Out of the 46 identified DMRs, 7 (6 for triclosan) encompassed imprinted genes (APC, FOXG1, GNAS, GNASAS, MIR886, PEG10, SGCE), which represented a significant enrichment. Other identified DMRs encompassed genes encoding proteins responsible for cell signaling, transmembrane transport, cell adhesion, inflammatory, apoptotic and immunological response, genes encoding transcription factors, histones, tumor suppressors, genes involved in tumorigenesis and several cancer risk biomarkers. Mediation analysis suggested that placental cell heterogeneity may partly explain these associations. This is the first study describing the genome-wide modifications of placental DNA methylation associated with pregnancy exposure to synthetic phenols or their precursors. Our results suggest that cell heterogeneity might mediate the effects of triclosan exposure on placental DNA methylation. Additionally, the enrichment of imprinted genes within the DMRs suggests mechanisms by which certain exposures, mainly to triclosan, could affect fetal development., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

A variety of specialty fibers such as no-core fiber (NCF) have already been studied to reveal their sensing abilities. In this work, we investigate a specialty fiber, square-core fiber, for temperature and strain sensing. A simple single-mode-multimode-single-mode (SMS) fiber sensor was fabricated, consisting of a 30-cm-long square-core fiber. The experimental results indicate that the maximal wavelength-temperature and wavelength-strain sensitivities are -15.3 pm/∘C and -1.5 pm/με, respectively, while the maximal power-temperature and power-strain sensitivities are 0.0896 dBm/∘C and 0.0756 dBm/με. Analysis of the results suggests that the fiber sensor has the potential to be used as a high-sensitivity temperature sensor with a low strain sensitivity.

To guarantee quality standards for the industry, surface properties, particularly those of roughness, must be considered in many areas of application. Today, several methods are available on the market, but some damage the surface to be tested as they measure it by contact. A non-contact method for the precise estimation of sub-micron roughness values is presented, which can be used as an extension of existing roughness measurement techniques to improve them further considering the depolarized light reflected by the sample. This setup is based on a Michelson interferometer, and by introducing a quarter-wave plate on a half part of the reference mirror, the surface roughness can be directly derived by measuring the fringe contrasts. This article introduces a simple model describing the intensity distortions resulting from the microscopic roughness in divided interferograms when considering depolarization. This work aimed to extend the measurement range of the technique developed in a previous work, in which depolarization effects are taken into account. For verification, the experimental results were compared with the fringe contrast technique, which does not consider the depolarization of the scattered light, especially regarding the extended wavelength interval, highlighting the limits of the technique. In addition, simulations of the experiments are presented. For comparison, the reference values of the sample roughness were also generated by measurements with a stylus profiler.

In this study, the response of fiber Bragg gratings (FBGs) embedded in cast aluminum parts under thermal and mechanical load were investigated. Several types of FBGs in different types of fibers were used in order to verify general applicability. To monitor a temperature-induced strain, an embedded regenerated FBG (RFBG) in a cast part was placed in a climatic chamber and heated up to 120 ∘C within several cycles. The results show good agreement with a theoretical model, which consists of a shrink-fit model and temperature-dependent material parameters. Several cast parts with different types of FBGs were machined into tensile test specimens and tensile tests were executed. For the tensile tests, a cyclic procedure was chosen, which allowed us to distinguish between the elastic and plastic deformation of the specimen. An analytical model, which described the elastic part of the tensile test, was introduced and showed good agreement with the measurements. Embedded FBGs - integrated during the casting process - showed under all mechanical and thermal load conditions no hysteresis, a reproducible sensor response, and a high reliable operation, which is very important to create metallic smart structures and packaged fiber optic sensors for harsh environments.

Biomaterials play an increasing role in clinical applications and regenerative medicine. A perfectly designed biomaterial should restore the function of damaged tissue without triggering an undesirable immune response, initiate self-regeneration of the surrounding tissue and gradually degrade after implantation. The immune system is well recognized to play a major role in influencing the biocompatibility of implanted medical devices. To obtain a better understanding of the effects of biomaterials on the immune response, we have developed a highly sensitive novel test system capable of examining changes in the immune system by biomaterial. Here, we evaluated for the first time the immunopeptidome, a highly sensitive system that reflects cancer transformation, virus or drug influences and passes these cellular changes directly to T cells, as a test system to examine the effects of contact with materials. Since monocytes are one of the first immune cells reacting to biomaterials, we have tested the influence of different materials on the immunopeptidome of the monocytic THP-1 cell line. The tested materials included stainless steel, aluminum, zinc, high-density polyethylene, polyurethane films containing zinc diethyldithiocarbamate, copper, and zinc sulfate. The incubation with all material types resulted in significantly modulated peptides in the immunopeptidome, which were material-associated. The magnitude of induced changes in the immunopeptidome after the stimulation appeared comparable to that of bacterial lipopolysaccharides (LPS). The source proteins of many detected peptides are associated with cytotoxicity, fibrosis, autoimmunity, inflammation, and cellular stress. Considering all tested materials, it was found that the LPS-induced cytotoxicity-, inflammation- and cellular stress-associated HLA class I peptides were mainly induced by aluminum, whereas HLA class II peptides were mainly induced by stainless steel. These findings provide the first insights into the effects of biomaterials on the immunopeptidome. A more thorough understanding of these effects may enable the design of more biocompatible implant materials using in vitro models in future. Such efforts will provide a deeper understanding of possible immune responses induced by biomaterials such as fibrosis, inflammation, cytotoxicity, and autoimmune reactions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Ghosh, Hartmann, Jakobi, März, Bichmann, Freudenmann, Mühlenbruch, Segan, Rammensee, Schneiderhan-Marra, Shipp, Stevanović and Joos.)

Ectomycorrhizal fungi (EMF) grow as saprotrophs in soil and interact with plants, forming mutualistic associations with roots of many economically and ecologically important forest tree genera. EMF ensheath the root tips and produce an extensive extramatrical mycelium for nutrient uptake from the soil. In contrast to other mycorrhizal fungal symbioses, EMF do not invade plant cells but form an interface for nutrient exchange adjacent to the cortex cells. The interaction of roots and EMF affects host stress resistance but uncovering the underlying molecular mechanisms is an emerging topic. Here, we focused on local and systemic effects of EMF modulating defenses against insects or pathogens in aboveground tissues in comparison with arbuscular mycorrhizal induced systemic resistance. Molecular studies indicate a role of chitin in defense activation by EMF in local tissues and an immune response that is induced by yet unknown signals in aboveground tissues. Volatile organic compounds may be involved in long-distance communication between below- and aboveground tissues, in addition to metabolite signals in the xylem or phloem. In leaves of EMF-colonized plants, jasmonate signaling is involved in transcriptional re-wiring, leading to metabolic shifts in the secondary and nitrogen-based defense metabolism but cross talk with salicylate-related signaling is likely. Ectomycorrhizal-induced plant immunity shares commonalities with systemic acquired resistance and induced systemic resistance. We highlight novel developments and provide a guide to future research directions in EMF-induced resistance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Dreischhoff, Das, Jakobi, Kasper and Polle.)

Background: We recently described increased NFATc1, IRF4, and NIP45 messenger RNA (mRNA) expression in peripheral blood mononuclear cells (PBMCs) of asthmatic children and adults with multiple allergies., Objective: NFATc2 has been described to associate with IRF4 to induce interleukin-4, and to be inhibited by T-bet. Here, we analyzed the role of NFATc2 in asthmatic children and adults., Methods: PBMCs were isolated from the blood of control of asthmatics subjects. Some PBMCs were analyzed untreated and some cultured with and without phytohemagglutinin. Then, RNA was extracted from the cells and cytokines were measured in the supernatants via enzyme-linked immunosorbent assay or multiplex analysis. RNA was then reverse-transcribed and NFATc1, NFATC2, IRF4, and T-bet mRNA were analyzed by real-time polymerase chain reaction. In addition, in peripheral blood cells, NFATc2 expression was analyzed, in a population of asthmatic children and adults from the Asthma BRIDGE study., Results: In addition to NFATc1 and NIP45, also NFATc2 was found upregulated in PBMCs and peripheral blood cells from asthmatic children and adults with allergic asthma. Moreover, NFATc1 directly correlated with lymphocytes number whereas NFATc2 correlated with peripheral eosinophilia in asthma., Conclusions: In addition to NFATc1 and NIP45, NFATc2 was found upregulated in asthma. Moreover, NFATc1 mRNA correlated with lymphocytes both in control and asthma, and NFATC1 and NFATc2 mRNA showed a direct correlation with eosinophils in controls but not in asthma, indicating that NFATc1 is associated with lymphocytes and not eosinophils in asthma., Clinical Significance: Targeting NFATc2 in T lymphocytes might ameliorate the allergic phenotype in asthmatic subjects., (© 2020 The Authors. Immunity, Inflammation and Disease published by John Wiley & Sons Ltd.)

Parkinson's disease-associated kinase LRRK2 has been linked to IFN type II (IFN-γ) response in infections and to dopaminergic neuronal loss. However, whether and how LRRK2 synergizes with IFN-γ remains unclear. In this study, we employed dopaminergic neurons and microglia differentiated from patient-derived induced pluripotent stem cells carrying LRRK2 G2019S, the most common Parkinson's disease-associated mutation. We show that IFN-γ enhances the LRRK2 G2019S-dependent negative regulation of AKT phosphorylation and NFAT activation, thereby increasing neuronal vulnerability to immune challenge. Mechanistically, LRRK2 G2019S suppresses NFAT translocation via calcium signaling and possibly through microtubule reorganization. In microglia, LRRK2 modulates cytokine production and the glycolytic switch in response to IFN-γ in an NFAT-independent manner. Activated LRRK2 G2019S microglia cause neurite shortening, indicating that LRRK2-driven immunological changes can be neurotoxic. We propose that synergistic LRRK2/IFN-γ activation serves as a potential link between inflammation and neurodegeneration in Parkinson's disease.

Absolute distance measurement is a field of research with a large variety of applications. Laser triangulation is a well-tested and developed technique using geometric relations to calculate the absolute distance to an object. The advantages of laser triangulation include its simple and cost-effective setup with yet a high achievable accuracy and resolution in short distances. A main problem of the technology is that even small changes of the optomechanical setup, e.g., due to thermal expansion, lead to significant measurement errors. Therefore, in this work, we introduce an optical setup containing only a beam splitter and a mirror, which splits the laser into a measurement beam and a reference beam. The reference beam can then be used to compensate for different error sources, such as laser beam dithering or shifts of the measurement setup due to the thermal expansion of the components. The effectiveness of this setup is proven by extensive simulations and measurements. The compensation setup improves the deviation in static measurements by up to 75%, whereas the measurement uncertainty at a distance of 1 m can be reduced to 85 μm. Consequently, this compensation setup can improve the accuracy of classical laser triangulation devices and make them more robust against changes in environmental conditions.

It has been widely shown that biomaterial surface topography can modulate host immune response, but a fundamental understanding of how different topographies contribute to pro-inflammatory or anti-inflammatory responses is still lacking. To investigate the impact of surface topography on immune response, we undertook a systematic approach by analyzing immune response to eight grades of medical grade polyurethane of increasing surface roughness in three in vitro models of the human immune system. Polyurethane specimens were produced with defined roughness values by injection molding according to the VDI 3400 industrial standard. Specimens ranged from 0.1 μ m to 18  μ m in average roughness (Ra), which was confirmed by confocal scanning microscopy. Immunological responses were assessed with THP-1-derived macrophages, human peripheral blood mononuclear cells (PBMCs), and whole blood following culture on polyurethane specimens. As shown by the release of pro-inflammatory and anti-inflammatory cytokines in all three models, a mild immune response to polyurethane was observed, however, this was not associated with the degree of surface roughness. Likewise, the cell morphology (cell spreading, circularity, and elongation) in THP-1-derived macrophages and the expression of CD molecules in the PBMC model on T cells (HLA-DR and CD16), NK cells (HLA-DR), and monocytes (HLA-DR, CD16, CD86, and CD163) showed no influence of surface roughness. In summary, this study shows that modifying surface roughness in the micrometer range on polyurethane has no impact on the pro-inflammatory immune response. Therefore, we propose that such modifications do not affect the immunocompatibility of polyurethane, thereby supporting the notion of polyurethane as a biocompatible material., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2020 Sören Segan et al.)

In this study, the response of regenerated fiber Bragg gratings (RFGBs) to axial forces was investigated in a temperature range from room temperature to 900 °C. For the first time, the transition from pure elastic to viscoelastic behavior around 700 °C of a standard SMF28 optical fiber was measured with an inscribed RFBG. An elastic model with linear temperature dependencies of Young's modulus and Poisson's ratio was established, and showed good agreement with the measurements up to temperatures of ∼500 °C. In the temperature range up to 900 °C, the RFBG response could be well described with a simple, single-material approach and a Burgers model that consists of a Kelvin and a Maxwell part. Based on the elastic parameter of the Maxwell part, the temperature-dependent force sensitivity of the RFBG was determined, and it showed a linear decrease in the range from room temperature to ∼500 °C, constant values in the range between ∼500 °C and ∼600 °C, and a strong increase at higher temperatures. While fulfilling the condition to operate in the elastic domain of the silica fiber, the investigations demonstrate that RFBGs can be used as force sensors up to temperatures of ∼600 °C - the range in which temperature-dependent force sensitivities have to be considered. The temperature-dependent parameters of the effective single-material model (elastic and viscoelastic part) are essential to describe the effective mechanical behavior of the optical fiber at high temperatures.

Background: We previously showed that the bacterial lipopeptide Pam 3 Cys-Ser-Ser, meanwhile established as a toll-like receptor (TLR) 1/2 ligand, acts as a strong adjuvant for the induction of virus specific CD8 + T cells in mice, when covalently coupled to a synthetic peptide., Case Presentation: We now designed a new water-soluble synthetic Pam 3 Cys-derivative, named XS15 and characterized it in vitro by a TLR2 NF-κB luciferase reporter assay. Further, the capacity of XS15 to activate immune cells and stimulate peptide-specific CD8 + T and NK cells by 6-sulfo LacNAc + monocytes was assessed by flow cytometry as well as cytokine induction using immunoassays. The induction of a functional immune response after vaccination of a volunteer with viral peptides was assessed by ELISpot assay and flow cytometry in peripheral blood cells and infiltrating cells at the vaccination site, as well as by immunohistochemistry and imaging. XS15 induced strong ex vivo CD8 + and T H 1 CD4 + responses in a human volunteer upon a single injection of XS15 mixed to uncoupled peptides in a water-in-oil emulsion (Montanide™ ISA51 VG). A granuloma formed locally at the injection site containing highly activated functional CD4 + and CD8 + effector memory T cells. The total number of vaccine peptide-specific functional T cells was experimentally assessed and estimated to be 3.0 × 10 5 in the granuloma and 20.5 × 10 6 in peripheral blood., Conclusion: Thus, in one volunteer we show a granuloma forming by peptides combined with an efficient adjuvant in a water-in-oil-emulsion, inducing antigen specific T cells detectable in circulation and at the vaccination site, after one single vaccination only. The ex vivo T cell responses in peripheral blood were detectable for more than one year and could be strongly boosted by a second vaccination. Hence, XS15 is a promising adjuvant candidate for peptide vaccination, in particular for tumor peptide vaccines in a personalized setting.

The characteristics of a surface, particularly the roughness, play an important role in different fields of the industry and have to be considered to ensure quality standards. Currently, there are numerous sophisticated methods for measuring surface roughness but plenty of them cause long-term damage because they are in contact with the sample. This article presents a non-contact method to accurately determine small surface roughnesses resulting from the consideration of the depolarization effects caused by the rough surface. This technique can be applied as an extension in various roughness measurements and improves the approach of Chandley's technique, which does not take into account the depolarization of the light scattered by the sample. The experimental setup and the measurements are easy to perform. The essential component is a quarter wave plate, which is incorporated into a Michelson interferometer. With the resulting two different contrasts and the recorded intensities of the sample and the reference mirror, the surface roughness can be estimated straightforwardly. This article details the theoretical approach, followed by the experimental results and the corresponding uncertainties. The experimental results are compared with Chandley's method. In order to have reference roughness values of the samples, measurements with a stylus profilometer and with a confocal microscope are performed and compared.

Continuous monitoring of biomarkers in a quantitative manner at point-of-care settings can advance early diagnosis in medicine. Contact lenses offer a minimally-invasive platform to continuously detect biomarkers in tear fluid. Microfluidic components as lab-on-a-chip technology have the potential to transform contact lenses into fully-integrated multiplexed sensing devices. Here, simple and complex microchannels are created in scleral lenses that perform microfluidic operations via capillary action. The engraving of microchannels in scleral lenses were performed by laser micromilling, where a predictive computational model was developed to simulate the effect of laser power and exposure time on polymer behavior. Experimentally varying the CO 2 laser power (1.2-3.6 W) and speed (38-100 mm s -1 ) allowed the micromilling of concave microchannels with groove depths of 10-240 μm and widths of 35-245 μm on polymetric substrates. The demonstrated laser micromilled circuitry in scleral lenses included linear channels, T/Y junctions, multiplexed arrays, mixers, and spiral channels, as well as serially organized multicomponent channels. Capillary forces acting in the microchannels allowed flowing rhodamine dye within the microfluidic components, which was visualized by optical microscopy in reflection and transmission modes simultaneously. The developed microfluidic components in scleral lenses may enable tear sampling, storage, analysis, and multiplexed detection capabilities for continuous monitoring applications., Competing Interests: The authors declare no competing financial interests., (This journal is © The Royal Society of Chemistry.)

A real-time, dual-sensitive shearography system using a single-wavelength laser was developed for simultaneous and dynamic in-plane and out-of-plane strain measurements. The shearography system is capable of measuring crack-tip deformation fields quantitatively. A spatial multiplexing technique based on Fourier transform is employed for simultaneous and dynamic multi-component phase retrieval. Two slit spatial filters and a common-path shearing interferometer are used to obtain an improved phase quality for crack-tip deformation measurements. Mode-I fracture experiments under three-point bending were conducted to validate the feasibility and the capability of this method.

This paper describes a dual-directional shearography system to address the issue of two-dimensional characterization of the surface strain. A common-path configuration coupled with an additional light path is used to provide the shearing in two directions. One of the three interfering beams is shared by both directional shearograms to improve the light efficiency and enhance the robustness of the system. The two directional shearograms are carried by different spatial carriers to distinguish one from the other. The spatial carrier is introduced by the single-aperture-lens Wollaston prism configuration. Rather than the conventional method in which the aperture is fixed at the front focal point of the imaging lens, a general case is considered by introducing a variable distance between the aperture and the imaging lens. The influence of the aperture-lens distance on the spatial carrier is then analyzed, which enables the separate control of the shearing amount and the spatial carrier. Two types of dual-directional shearography are presented to demonstrate the feasibility and the flexibility of the system. Type I is the simultaneous dual lateral shearography in orthogonal directions, and Type II is the simultaneous lateral and radial shearography. The spatial carrier introduced by the single-aperture-lens Wollaston prism configuration is discussed, and a configuration in which the Wollaston prism and the aperture are located at different sides of the lens is recommended for further shearography applications.

Measurement systems for online nondestructive full-field three-dimensional (3D) displacement based on the single-shot and multiplexing techniques attract more and more interest, especially throughout the manufacturing industries. This paper proposes an accurate and easy-to-implement method based on an electronic speckle pattern interferometer (ESPI) with single illumination-detection path to realize the online nondestructive full-field 3D displacement measurement. The simple and compact optical system generates three different sensitivity vectors to enable the evaluation of the three orthogonal displacement components. By applying the spatial carrier phase-shifting technique, the desired information can be obtained in real time. The theoretical analysis and the measurement results have proven the feasibility of this ESPI system and quantified its relative measurement error.

A temporal electronic speckle pattern interferometry (ESPI) system is proposed for in-plane rotation measurement. The relationship between the rotation angle and the phase change distribution is established and the rotation direction is indicated by the sign of the partial differential of the phase change distribution. Temporal phase modulation is applied in the proposed symmetric illumination ESPI system. The phase is recovered by the temporal intensity analysis method which uses the temporal evolution history of the light intensity. The system can perform dynamic measurements and provide results in off-line real-time. Preliminary experiments were carried out with a continuously rotating target to show the feasibility and the dynamic feature of the temporal ESPI system. At present, the mean absolute error of the experiment is 0.39 arcsec.

In recent years, the observation of the behaviour of components during the production process and over their life cycle is of increasing importance. Structural health monitoring, for example of carbon composites, is state-of-the-art research. The usage of Fibre Bragg Gratings (FBGs) in this field is of major advantage. Another possible area of application is in foundries. The internal state of melts during the solidification process is of particular interest. By using embedded FBGs, temperature and stress can be monitored during the process. In this work, FBGs were embedded in aluminium alloys in order to observe the occurring strain. Two different FBG positions were chosen in the mould in order to compare its dependence. It was shown that FBGs can withstand the solidification process, although a compression in the range of one percent was measured, which is in agreement with the literature value. Furthermore, different lengths of the gratings were applied, and it was shown that shorter gratings result in more accurate measurements. The obtained results prove that FBGs are applicable as sensors for temperatures up to 740 °C., Competing Interests: The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses or interpretation of data; in the writing of the manuscript; nor in the decision to publish the results.

Soil-dwelling Streptomyces bacteria such as S.coelicolor have to constantly adapt to the nitrogen (N) availability in their habitat. Thus, strict transcriptional and post-translational control of the N-assimilation is fundamental for survival of this species. GlnR is a global response regulator that controls transcription of the genes related to the N-assimilation in S. coelicolor and other members of the Actinomycetales. GlnR represents an atypical orphan response regulator that is not activated by the phosphorylation of the conserved aspartate residue (Asp 50). We have applied transcriptional analysis, LC-MS/MS analysis and electrophoretic mobility shift assays (EMSAs) to understand the regulation of GlnR in S. coelicolor M145. The expression of glnR and GlnR-target genes was revisited under four different N-defined conditions and a complex N-rich condition. Although, the expression of selected GlnR-target genes was strongly responsive to changing N-concentrations, the glnR expression itself was independent of the N-availability. Using LC-MS/MSanalysis we demonstrated that GlnR was post-translationally modified. The post-translational modifications of GlnR comprise phosphorylation of the serine/threonine residues and acetylation of lysine residues. In the complex N-rich medium GlnR was phosphorylated on six serine/threonine residues and acetylated on one lysine residue. Under defined N-excess conditions only two phosphorylated residues were detected whereas under defined N-limiting conditions no phosphorylation was observed. GlnR phosphorylation is thus clearly correlated with N-rich conditions. Furthermore, GlnR was acetylated on four lysine residues independently of the N-concentration in the defined media and on only one lysine residue in the complex N-rich medium. Using EMSAs we demonstrated that phosphorylation inhibited the binding of GlnR to its targets genes, whereas acetylation had little influence on the formation of GlnR-DNA complex. This study clearly demonstrated that GlnR DNA-binding affinity is modulated by post-translational modifications in response to changing N-conditions in order to elicit a proper transcriptional response to the latter.

The unfolded protein response (UPR), a conserved eukaryotic signaling pathway to ensure protein homeostasis in the endoplasmic reticulum (ER), coordinates biotrophic development in the corn smut fungus Ustilago maydis. Exact timing of UPR activation is required for virulence and presumably connected to the elevated expression of secreted effector proteins during infection of the host plant Zea mays. In the baker's yeast Saccharomyces cerevisiae, expression of UPR target genes is induced upon binding of the central regulator Hac1 to unfolded protein response elements (UPREs) in their promoters. While a role of the UPR in effector secretion has been described previously, we investigated a potential UPR-dependent regulation of genes encoding secreted effector proteins. In silico prediction of UPREs in promoter regions identified the previously characterized effector genes pit2 and tin1-1, as bona fide UPR target genes. Furthermore, direct binding of the Hac1-homolog Cib1 to the UPRE containing promoter fragments of both genes was confirmed by quantitative chromatin immunoprecipitation (qChIP) analysis. Targeted deletion of the UPRE abolished Cib1-dependent expression of pit2 and significantly affected virulence. Furthermore, ER stress strongly increased Pit2 expression and secretion. This study expands the role of the UPR as a signal hub in fungal virulence and illustrates, how biotrophic fungi can coordinate cellular physiology, development and regulation of secreted virulence factors.