Assessment of centre national d'Études spatiales real-time ionosphere maps in instantaneous precise real-time kinematic positioning over medium and long baselines

Precise real-time kinematic (RTK) Global Navigation Satellite System (GNSS) positioning requires fixing integer ambiguities after a short initialization time. Originally, it was assumed that it was only possible at a relatively short distance from a reference station (<10 km), because otherwise the atmospheric effects prevent effective ambiguity fixing. Nowadays, through the use of VRS, MAC, or FKP corrections, the distances to the closest reference station have been increased to around 35 km. However, the baselines resolved in real time are not as far as in the case of static positioning. Further extension of the baseline requires the use of an ionosphere-weighted model with ionospheric delay corrections available in real time. This solution is now possible thanks to the Radio Technical Commission for Maritime (RTCM) stream of SSR corrections from, for example, Centre National d’Études Spatiales (CNES), the first analysis center to provide it in the context of the International GNSS Service. Then, ionospheric delays are treated as pseudo-observations that have a priori values from the CLK RTCM stream. Additionally, satellite orbit and clock errors are properly considered using space-state representation (SSR) real-time radial, along-track, and cross-track corrections. The following paper presents the initial results of such RTK positioning. Measurements were performed in various field conditions reflecting realistic scenarios that could have been experienced by actual RTK users. We have shown that the assumed methodology was suitable for single-epoch RTK positioning with up to 82 km baseline in solar minimum (30 March 2019) mid and high latitude (Olsztyn, Poland) conditions. We also confirmed that it is possible to obtain a rover position at the level of a few centimeters of precision. Finally, the possibility of using other newer experimental IGS RT Global Ionospheric Maps (GIMs), from Chinese Academy of Sciences (CAS) and Universitat Politècnica de Catalunya (UPC) among CNES, is discussed in terms of their recent performance in the ionospheric delay domain.Peer ReviewedPostprint (published version

Ionospheric tomographic common clock model of undifferenced uncombined GNSS measurements

This is a post-peer-review, pre-copyedit version of an article published in Journal of geodesy. The final authenticated version is available online at: http://dx.doi.org/10.1007/s00190-021-01568-8In this manuscript, we introduce the Ionospheric Tomographic Common Clock (ITCC) model of undifferenced uncombined GNSS measurements. It is intended for improving the Wide Area precise positioning in a consistent and simple way in the multi-GNSS context, and without the need of external precise real-time products. This is the case, in particular, of the satellite clocks, which are estimated at the Wide Area GNSS network Central Processing Facility (CPF) referred to the reference receiver one; and the precise realtime ionospheric corrections, simultaneously computed under a voxel-based tomographic model with satellite clocks and other geodetic unknowns, from the uncombined and undifferenced pseudoranges and carrier phase measurements at the CPF from the Wide Area GNSS network area. The model, without fixing the carrier phase ambiguities for the time being (just constraining them by the simultaneous solution of both ionospheric and geometric components of the uncombined GNSS model), has been successfully applied and assessed against previous precise positioning techniques. This has been done by emulating real-time conditions for Wide Area GPS users during 2018 in Poland.Peer ReviewedPostprint (published version

Improved MSTID modelling and impact on precise GNSS processing

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Direct MSTID mitigation in precise GPS processing

This is the peer reviewed version of the following article: Hernandez, M., Wielgosz, P., Paziewski, J., Krypiak-Gregorczyk, A., Krukowska, M., Stepniak, K., Kaplon, J., Hadas, T., Sosnica, K., Bosy, J., Orús, R., Monte, E., Yang, H., Garcia-Rigo, A., Olivares-Pulido, G. Direct MSTID mitigation in precise GPS processing. "Radio science", Març 2017, vol. 52, núm. 3, p. 321-337, which has been published in final form at http://onlinelibrary.wiley.com/doi/10.1002/2016RS006159/abstract. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-ArchivingIn this paper, the authors summarize a simple and efficient approach developed to mitigate the problem in precise GNSS positioning originated by the most frequent ionospheric wave signatures: the Medium Scale Travelling Ionospheric Disturbances (MSTIDs). The direct GNSS Ionospheric Interferometry technique (hereinafter dGII), presented in this paper, is applied for correcting MSTID effects on precise Real Time Kinematic (RTK) and tropospheric determination. It consists of the evolution of the former climatic Differential Delay Mitigation Model for MSTIDs (DMTID), for real-time conditions, using ionospheric data from a single permanent receiver only. The performance is demonstrated with networks of GNSS receivers in Poland, treated as users under real-time conditions, during two representative days in winter and summer seasons (days 353 and 168 of year 2013). In range domain, dGII typically reduces the ionospheric delay error up to 10-90% of the value when the MSTID mitigation model is not applied. The main dGII impact on precise positioning is that we can obtain reliable RTK position faster. In particular the ASR (ambiguity success rate) parameter increases, from 74% to 83%, with respect to the original uncorrected observations. The average of time to first fix is shortened from 30s to 13s. The improvement in troposphere estimaton, due to any potential impact of the MSTID mitigation model, was most difficult to demonstrate.Peer ReviewedPostprint (author's final draft

Design and In Vitro Evaluation of a Cytotoxic Conjugate Based on the Anti-HER2 Affibody Fused to the Fc Fragment of IgG1

In our previous work we demonstrated that a small protein called affibody can be used for a cytotoxic conjugate development. The anti-HER2 affibody was armed with one moiety of a highly potent auristatin E and specifically killed HER2-positive cancer cells with a nanomolar IC50. The aim of this study was to improve the anti-HER2 affibody conjugate by increasing its size and the number of conjugated auristatin molecules. The affibody was fused to the Fc fragment of IgG1 resulting in a dimeric construct with the molecular weight of 68 kDa, referred to as ZHER2:2891-Fc, ensuring its prolonged half-life in the blood. Due to the presence of four interchain cysteines, the fusion protein could carry four drug molecules. Notably, the in vitro tests of the improved anti-HER2 conjugate revealed that it exhibits the IC50 of 130 pM for the HER2-positive SK-BR-3 cells and 98 nM for the HER2-negative MDA-MB-231 cells. High efficacy and specificity of the auristatin conjugate based on ZHER2:2891-Fc indicate that this construct is suitable for further in vivo evaluation

Correction: Serwotka-Suszczak, A. M. et al. A Conjugate Based on Anti-HER2 Diaffibody and Auristatin E Targets HER2-Positive Cancer Cells. <em>Int. J. Mol. Sci.</em> 2017, <em>18</em>, 401

It has been brought to our attention that the affiliation of Dr. Jerzy Pieczykolan at the time when he was responsible for the work described in the paper [...

A Conjugate Based on Anti-HER2 Diaffibody and Auristatin E Targets HER2-Positive Cancer Cells

Antibody-drug conjugates (ADCs) have recently emerged as efficient and selective cancer treatment therapeutics. Currently, alternative forms of drug carriers that can replace monoclonal antibodies are under intensive investigation. Here, a cytotoxic conjugate of an anti-HER2 (Human Epidermal Growth Factor Receptor 2) diaffibody with monomethyl-auristatin E (MMAE) is proposed as a potential anticancer therapeutic. The anti-HER2 diaffibody was based on the ZHER2:4 affibody amino acid sequence. The anti-HER2 diaffibody has been expressed as a His-tagged protein in E. coli and purified by Ni-nitrilotriacetyl (Ni-NTA) agarose chromatography. The molecule was properly folded, and the high affinity and specificity of its interaction with HER2 was confirmed by surface plasmon resonance (SPR) and flow cytometry, respectively. The (ZHER2:4)2DCS-MMAE conjugate was obtained by coupling the maleimide group linked with MMAE to cysteines, which were introduced in a drug conjugation sequence (DCS). Cytotoxicity of the conjugate was evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide MTT assay and the xCELLigence Real-Time Cell Analyzer. Our experiments demonstrated that the conjugate delivered auristatin E specifically to HER2-positive tumor cells, which finally led to their death. These results indicate that the cytotoxic diaffibody conjugate is a highly potent molecule for the treatment of various types of cancer overexpressing HER2 receptors

PIOM-FIPP: TN1

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N-glycosylation acts as a switch for FGFR1 trafficking between the plasma membrane and nuclear envelope

Abstract Fibroblast growth factor receptor 1 (FGFR1) is a heavily N-glycosylated cell surface receptor tyrosine kinase that transmits signals across the plasma membrane, in response to fibroblast growth factors (FGFs). Balanced FGF/FGFR1 signaling is crucial for the development and homeostasis of the human body, and aberrant FGFR1 is frequently observed in various cancers. In addition to its predominant localization to the plasma membrane, FGFR1 has also been detected inside cells, mainly in the nuclear lumen, where it modulates gene expression. However, the exact mechanism of FGFR1 nuclear transport is still unknown. In this study, we generated a glycosylation-free mutant of FGFR1, FGFR1.GF, and demonstrated that it is localized primarily to the nuclear envelope. We show that reintroducing N-glycans into the D3 domain cannot redirect FGFR1 to the plasma membrane or exclude the receptor from the nuclear envelope. Reestablishment of D2 domain N-glycans largely inhibits FGFR1 accumulation in the nuclear envelope, but the receptor continues to accumulate inside the cell, mainly in the ER. Only the simultaneous presence of N-glycans of the D2 and D3 domains of FGFR1 promotes efficient transport of FGFR1 to the plasma membrane. We demonstrate that while disturbed FGFR1 folding results in partial FGFR1 accumulation in the ER, impaired FGFR1 secretion drives FGFR1 trafficking to the nuclear envelope. Intracellular FGFR1.GF displays a high level of autoactivation, suggesting the presence of nuclear FGFR1 signaling, which is independent of FGF. Using mass spectrometry and proximity ligation assay, we identified novel binding partners of the nuclear envelope-localized FGFR1, providing insights into its cellular functions. Collectively, our data define N-glycosylation of FGFR1 as an important regulator of FGFR1 kinase activity and, most importantly, as a switchable signal for FGFR1 trafficking between the nuclear envelope and plasma membrane, which, due to spatial restrictions, shapes FGFR1 interactome and cellular function. Video Abstrac