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Chimeric Antigen Receptor (CAR) T cells are effective at targeting tumor cells, particularly in hematological malignancies. A safer, switchable modular system called RevCAR has been developed to address dangerous side-effects. This system includes RevCAR T cells, which cannot bind to targets on their own, and a bispecific target module (RevTM). The functionality of this system depends on the presence of RevTM, which acts as a safety switch. Additionally, different RevTMs can be used with the same T cells to target various antigens, providing greater flexibility. To effectively treat solid tumors, however, it is necessary to overcome the immunosuppressive tumor microenvironment. To address this, new RevTMs were created to target immune checkpoint PD-L1, which cancer cells often upregulate to evade the immune system. Our research demonstrated that these new RevTMs enable RevCAR T cells to specifically target and kill a wide range of PD-L1-expressing cells in both monolayer and 3D models. The RevCAR T cells also released pro-inflammatory cytokines and expressed activation markers after co-culture. Furthermore, we validated an AND-gated targeting approach that simultaneously targets a tumor-associated antigen (TAA) and an immune checkpoint. These findings suggest a promising new strategy for applying the RevCAR platform to the treatment of solid tumors.

Background Floodplains are biodiversity hotspots and provide numerous ecosystem services. In recent decades, however, 70-90% of Europe's floodplains have been structurally degraded. Accordingly, many (inter-)national programs aim to restore and protect floodplain ecosystems. The success of such measures also depends on the chemical contamination, especially of floodplain soils and sediments, which serve as sinks and sources for a variety of pollutants. In this study, we assess the current ecotoxicological status of a floodplain restoration site along the Main River (Frankfurt am Main, Germany) and estimate its development potential with respect to the influence of a local industrial plant and potential legacy contaminations. We therefore use in vitro effect-based methods (EBMs) testing for baseline toxicity, mutagenicity, dioxin-like and estrogenic activities, coupled with chemical analysis. Results Of all water bodies analyzed, the overall toxicity was highest in two flood depressions. In the respective water phase, estrogenic activities exceeded the environmental quality standard and sediment samples were positive for all tested endpoints. Chemical analysis of these sediments revealed high concentrations of polycyclic aromatic hydrocarbons. Soil samples from frequently flooded areas showed the highest mutagenic potential for both frameshift and point mutations with and without metabolic activation. The industrial effluent showed baseline toxic, mutagenic, and dioxin-like activities, that were highly diluted in the Main River. In turn, most of the sediment samples downstream of the industrial discharge showed significantly elevated baseline toxic, estrogenic and dioxin-like activities as well as increased chemical contamination. Conclusion Based on the results of this study, we rate the overall ecotoxicological status of a recently established tributary and groundwater-fed ponds as good, and identified two flood depressions near the Main River as hot spots of conta

T cells can be armed with chimeric antigen receptors (CARs) to target tumor cells, a strategy which has proven successful against some hematological cancers. To overcome some of the dangerous side-effects, a modular system termed RevCAR has been developed. Its components are the RevCAR T cells, which alone cannot bind to any targets, and a bispecific targeting molecule (RevTM). Since the presence of the RevTM is necessary for the system to be functional, it can be used as a safety switch. In addition, different RevTMs can be used with the same T cells to target different antigens, achieving more flexibility. In order to achieve a successful treatment of solid tumors, however, the immune suppression of the tumor microenvironment needs to be overcome. To do this, novel RevTMs were designed which target immune checkpoint molecules such as PD-L1, a pathway which is often exploited by cancer cells to block the immune response. We have successfully shown that, in the presence of these new RevTMs, RevCAR T cells are able to specifically target and kill a wide range of PD-L1-expressing cells, both in monolayer and 3D models. They also release pro-inflammatory cytokines and express activation markers after the co-culture. Furthermore, a combinatorial targeting approach of both a TAA and an immune checkpoint has been proven. Together, these results demonstrate a promising new strategy to apply the RevCAR platform to solid tumors.

The presence of tyre and road wear particles (TRWP) in the environment is an underestimated threat due to their potential impact on ecosystems and human health. However, their mode of action and potential impacts on aquatic ecosystems remain largely unknown. In the present study, we adopted a sediment exposure scenario to investigate the influence of sediment coming from an urban runoff sedimentation basin on the life cycle of Chironomus riparius. Targeted broad-spectrum chemical analysis helped to characterise the urban sediments and confirmed the significant contribution of contaminants from traffic (e.g. tyre wear contribution, Polycyclic Aromatic Hydrocarbons [PAHs], metals, tyre rubber additives). First-stage chironomid larvae were subjected to increasing concentrations of urban whole sediment. The results showed that exposure to this urban sediment influenced all measured endpoints. In vivo quantification of ROS showed that larvae exposed to the lowest concentration of contaminated sediment exhibited increased fluorescence. The contaminated sediment conditions increased mortality by almost 30 %, but this effect was surprisingly not concentration-dependent. Fertility decreased significantly and concentration-dependently. The results of the Mean Emergence Time (EmT50) and larval size showed an optimality curve. Furthermore, as a consequence of the effects on fitness, the Population Growth Rate (PGR) exhibited a significant decrease, which was concentration-dependent. Therefore, after a single generation, PGR calculation can be adopted as a sensitive tool to monitor pollution caused by complex matrices, i.e. composed of several contaminants. Our research highlights the importance of effective management of road runoff and underlines the need for further investigation to better understand the toxicity of TRWPs.

Given the ever-increasing number of time-domain astronomical surveys, employing robust, interpretative, and automated data-driven classification schemes is pivotal. Based on graph theory, we present new data-driven classification heuristics for spectral data. A spectral classification scheme of Type II supernovae (SNe II) is proposed based on the phase relative to the maximum light in the V band and the end of the plateau phase. We utilize a compiled optical data set that comprises 145 SNe and 1595 optical spectra in 4000-9000 angstrom. Our classification method naturally identifies outliers and arranges the different SNe in terms of their major spectral features. We compare our approach to the off-the-shelf UMAP manifold learning and show that both strategies are consistent with a continuous variation of spectral types rather than discrete families. The automated classification naturally reflects the fast evolution of Type II SNe around the maximum light while showcasing their homogeneity close to the end of the plateau phase. The scheme we develop could be more widely applicable to unsupervised time series classification or characterization of other functional data.

Contains fulltext : 291814.pdf (Publisher’s version ) (Open Access)

Contains fulltext : 291814.pdf (Publisher’s version ) (Open Access)

Chimeric antigen receptors (CARs) are used for redirection of T cells against tumor cells and have demonstrated remarkable therapeutic effects against some hematological cancers. The modular system termed RevCAR can overcome the dangerous side effects associated with conventional CAR T cell therapy, such as cytokine release syndrome. It is composed of RevCAR T cells and a bispecific molecule target module, termed RevTM. This system is highly safe since RevCAR T cell activity can be controlled by RevTM and immediately switched off if side effects are detected. It is also versatile, since the same RevCAR T cell can be directed to different tumor-associated antigens (TAA) simply by adding RevTMs with different specificities. However, to improve the effectiveness in the treatment of solid tumors, the immunosuppressive tumor microenvironment (TME) still needs to be addressed. For this purpose, we have developed novel RevTMs targeting immune checkpoint molecules such as PD-L1, which are often upregulated by cancer cells to escape the immune cells. We have demonstrated that our novel RevTMs can redirect RevCAR T cells to specifically kill cell lines expressing immune checkpoint molecules. In addition to this strategy, the combinatorial targeting of a TAA and an immune checkpoint is very promising strategy to overcome the immunosuppressive microenvironment of solid tumors and thus to improve the treatment outcome.

Engineered T cells that express chimeric antigen receptors (CARs) show a big potential in cancer immunological research. They are synthetic receptors that can target antigens independently from the MHC complex. However, most tumor associated antigens are not only expressed on the tumor site but also on healthy tissues. To switch off CAR T cells in case of on-target/off-tumor effects occur, an adaptor CAR platform, called the universal CAR (UniCAR), was developed. In contrast to conventional CAR systems this platform consists of the engineered T cells and a linker molecule, called target module (TM), that is both binding to the target antigen and recognized by the UniCAR T cell. With this approach there is the possibility to switch the system on and off when needed. Fibroblast activation protein (FAP) is a protein of high interest in cancer research as it is upregulated in most of epithelial cancers but shows a comparably low expression in healthy tissues. It is shown to be expressed on cancer associated fibroblasts and therefore plays an important role in the tumor microenvironment where it has pro-tumorigenic activity both in the FAP positive and surrounding cells. Given this, the goal of this work was to develop new TMs against FAP. To achieve such aim, we cloned two TM formats, either based on a single-chain variable fragment (scFv) or an IgG4 based backbone. The difference in the size of the backbones translates into different half-lifes and kinetics. These different TM formats can be used to customize UniCAR therapy according to the given circumstances. We were able to purify and perform various experiments with the mentioned TMs. We could show binding to the target cells using flow cytometry. We could also show that the TMs could be used to induce specific lysis in vitro on monolayer cells and spheroids. We were also able to show UniCAR T cell infiltration into the spheroids and their consequent activation. Moreover, in mouse models the TMs could be used for P

T-cells genetically modified to express chimeric antigen receptors (CARs) are playing a more and more important role in targeted cancer immunotherapy. However, these living drugs can also cause lifethreatening side effects. To overcome such limitations and improve the safety of CAR T-cell therapy, adaptor CAR platforms such as the universal CAR (UniCAR) have been developed. This platform consists of the UniCAR T-cell and a target module (TM) cross-linkage effector and tumor cells. Here, we have established a novel UniCAR system targeting Fibroblast Activation Protein (FAP) that is highly expressed in the tumor microenvironment of epithelial cancers and a marker for cancer-associated fibroblasts. For that, we constructed two novel FAP-directed TMs possessing different sizes and pharmacokinetic properties, in which one is based on a single-chain variable fragment (scFv), and the other is based on an IgG4 backbone. We have shown that both TMs were able to bind to FAP-expressing cells and redirect UniCAR T-cells in vitro to monolayer and spheroid target cells inducing effective killing. Furthermore, we could show infiltration and activation of T-cells in the spheroid setting. Using in vivo models, the TMs were proven to be suitable to be used for PET imaging showing FAP-specific accumulation at the tumor site. Moreover, the immunotherapeutic effect of UniCAR T-cells in combination with FAP TMs was demonstrated using mouse models. In conclusion, in this work, we could show that the two novel anti- FAP TMs prove to hold great theranostic potential for diagnostic imaging and immunotherapy.

Immunotherapy based on chimeric antigen receptor (CAR) T-cells has demonstrated remarkable therapeutic effects, particularly against some hematological cancers. A versatile adaptor CAR system called RevCAR, consisting of RevCAR T-cells and a bispecific target module (RevTM) has been developed to overcome severe side effects associated with conventional CAR T-cell therapy. As the activity of RevCAR T-cells can be steered based on the availability of RevTM, working as an on/off switch, the system can be immediately turned off if side effects occur. Furthermore, the RevCAR system is highly flexible, since the same RevCAR T-cell can be directed towards different tumor-associated antigens (TAA) simply by adding RevTMs with different specificities. However, the effectiveness of CAR T-cells against solid tumors remains limited particularly due to their immunosuppressive tumor microenvironment (TME). To overcome these hurdles, we have established a novel RevCAR system targeting immune checkpoint molecules such as PD-L1, which are frequently overexpressed by cancer cells to suppress immune responses. We have constructed novel RevTMs that can redirect RevCAR T-cells to kill tumor cells that express such immune checkpoint molecules. Furthermore, true AND gate tumor targeting was achieved by targeting a TAA in addition to PD-L1 in a combinatorial manner using our Dual-RevCAR system. In this way, targeting PD-L1 not only results in Dual-RevCAR T-cell activation but simultaneously blocks the immunosuppressive PD-L1/PD-1 axis. Altogether, we have turned an immunosuppressive marker into an immune-activating signal that might modulate the TME in a beneficial manner, showing promise for the development of an effective immunotherapy against solid tumors.

TheraSTAR – Project Overview

Background: Chimeric antigen receptor (CAR) T-cells are a promising approach in cancer immunotherapy, particularly for treating hematologic malignancies. Yet, their effectiveness is limited when tackling solid tumors, where immune cell infiltration and immunosuppressive tumor microenvironments (TME) are major hurdles. Fibroblast activation protein (FAP) is highly expressed on cancer-associated fibroblasts (CAFs) and various tumor cells, playing an important role in tumor growth and immunosuppression. Aiming to modulate the TME with increased clinical safety and effectiveness, we developed novel small and size-extended immunotheranostic UniCAR target modules (TMs) targeting FAP. Methods: The specific binding and functionality of the anti-FAP-scFv TM and the size extended anti-FAP-IgG4 TM were assessed using 2D and 3D in vitro models as well as in vivo. Their specific tumor accumulation and diagnostic potential was evaluated using PET studies after functionalization with a chelator and suitable radionuclide. Results: The anti-FAP-scFv and -IgG4 TMs effectively and specifically redirected UniCAR T-cells using 2D, 3D, and in vivo models. Moreover, a remarkably high and specific accumulation of radiolabeled FAP-targeting TMs at the tumor site of xenograft mouse models was observed. Conclusions: These findings demonstrate that the novel anti-FAP TMs are promising immunotheranostic tools to foster cancer imaging and treatment, paving the way for a more convenient, individualized, and safer treatment of cancer patients.

The cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitor palbociclib is an emerging cancer therapeutic that just recently gained Food and Drug Administration approval for treatment of estrogen receptor (ER)-positive, human epidermal growth factor receptor (Her)2-negative breast cancer in combination with the ER degrader fulvestrant. However, CDK4/6 inhibitors are not cancer-specific and may affect also other proliferating cells. Given the importance of T cells in antitumor defense, we studied the influence of palbociclib/fulvestrant on human CD3+ T cells and novel emerging T cell-based cancer immunotherapies. Palbociclib considerably inhibited the proliferation of activated T cells by mediating G0/G1 cell cycle arrest. However, after stopping the drug supply this suppression was fully reversible. In light of combination approaches, we further investigated the effect of palbociclib/fulvestrant on T cell-based immunotherapies by using a CD3-PSCA bispecific antibody or universal chimeric antigen receptor (UniCAR) T cells. Thereby, we observed that palbociclib clearly impaired T cell expansion. This effect resulted in a lower total concentration of interferon-g and tumor necrosis factor, while palbociclib did not inhibit the average cytokine release per cell. In addition, the cytotoxic potential of the redirected T cells was unaffected by palbociclib and fulvestrant. Overall, these novel findings may have implications for the design of treatment modalities combining CDK4/6 inhibition and T cell-based cancer immunotherapeutic strategies.

Immunotherapy based on chimeric antigen receptor (CAR) T-cells has demonstrated remarkable therapeutic effects, particularly against some hematological cancers. A versatile adaptor CAR system called RevCAR, consisting of RevCAR T-cells and a bispecific target module (RevTM) has been developed to overcome severe side effects associated with conventional CAR T-cell therapy. As the activity of RevCAR T-cells can be steered based on the availability of RevTM, working as an on/off switch, the system can be immediately turned off if side effects occur. Furthermore, the RevCAR system is highly flexible, since the same RevCAR T-cell can be directed towards different tumor-associated antigens (TAA) simply by adding RevTMs with different specificities. However, the effectiveness of CAR T-cells against solid tumors remains limited particularly due to their immunosuppressive tumor microenvironment (TME). To overcome these hurdles, we have established a novel RevCAR system targeting immune checkpoint molecules such as PD-L1, which are frequently overexpressed by cancer cells to suppress immune responses. We have constructed novel RevTMs that can redirect RevCAR T-cells to kill tumor cells that express such immune checkpoint molecules. Furthermore, true AND gate tumor targeting was achieved by targeting a TAA in addition to PD-L1 in a combinatorial manner using our Dual-RevCAR system. In this way, targeting PD-L1 not only results in Dual-RevCAR T-cell activation but simultaneously blocks the immunosuppressive PD-L1/PD-1 axis. Altogether, we have turned an immunosuppressive marker into an immune-activating signal that might modulate the TME in a beneficial manner, showing promise for the development of an effective immunotherapy against solid tumors.

T-cells genetically modified to express chimeric antigen receptors (CARs) are playing a more and more important role in targeted cancer immunotherapy. However, these living drugs can also cause lifethreatening side effects. To overcome such limitations and improve the safety of CAR T-cell therapy, adaptor CAR platforms such as the universal CAR (UniCAR) have been developed. This platform consists of the UniCAR T-cell and a target module (TM) cross-linkage effector and tumor cells. Here, we have established a novel UniCAR system targeting Fibroblast Activation Protein (FAP) that is highly expressed in the tumor microenvironment of epithelial cancers and a marker for cancer-associated fibroblasts. For that, we constructed two novel FAP-directed TMs possessing different sizes and pharmacokinetic properties, in which one is based on a single-chain variable fragment (scFv), and the other is based on an IgG4 backbone. We have shown that both TMs were able to bind to FAP-expressing cells and redirect UniCAR T-cells in vitro to monolayer and spheroid target cells inducing effective killing. Furthermore, we could show infiltration and activation of T-cells in the spheroid setting. Using in vivo models, the TMs were proven to be suitable to be used for PET imaging showing FAP-specific accumulation at the tumor site. Moreover, the immunotherapeutic effect of UniCAR T-cells in combination with FAP TMs was demonstrated using mouse models. In conclusion, in this work, we could show that the two novel anti- FAP TMs prove to hold great theranostic potential for diagnostic imaging and immunotherapy.

Chimeric antigen receptors (CARs) are used for redirection of T cells against tumor cells and have demonstrated remarkable therapeutic effects against some hematological cancers. The modular system termed RevCAR can overcome the dangerous side effects associated with conventional CAR T cell therapy, such as cytokine release syndrome. It is composed of RevCAR T cells and a bispecific molecule target module, termed RevTM. This system is highly safe since RevCAR T cell activity can be controlled by RevTM and immediately switched off if side effects are detected. It is also versatile, since the same RevCAR T cell can be directed to different tumor-associated antigens (TAA) simply by adding RevTMs with different specificities. However, to improve the effectiveness in the treatment of solid tumors, the immunosuppressive tumor microenvironment (TME) still needs to be addressed. For this purpose, we have developed novel RevTMs targeting immune checkpoint molecules such as PD-L1, which are often upregulated by cancer cells to escape the immune cells. We have demonstrated that our novel RevTMs can redirect RevCAR T cells to specifically kill cell lines expressing immune checkpoint molecules. In addition to this strategy, the combinatorial targeting of a TAA and an immune checkpoint is very promising strategy to overcome the immunosuppressive microenvironment of solid tumors and thus to improve the treatment outcome.

Engineered T cells that express chimeric antigen receptors (CARs) show a big potential in cancer immunological research. They are synthetic receptors that can target antigens independently from the MHC complex. However, most tumor associated antigens are not only expressed on the tumor site but also on healthy tissues. To switch off CAR T cells in case of on-target/off-tumor effects occur, an adaptor CAR platform, called the universal CAR (UniCAR), was developed. In contrast to conventional CAR systems this platform consists of the engineered T cells and a linker molecule, called target module (TM), that is both binding to the target antigen and recognized by the UniCAR T cell. With this approach there is the possibility to switch the system on and off when needed. Fibroblast activation protein (FAP) is a protein of high interest in cancer research as it is upregulated in most of epithelial cancers but shows a comparably low expression in healthy tissues. It is shown to be expressed on cancer associated fibroblasts and therefore plays an important role in the tumor microenvironment where it has pro-tumorigenic activity both in the FAP positive and surrounding cells. Given this, the goal of this work was to develop new TMs against FAP. To achieve such aim, we cloned two TM formats, either based on a single-chain variable fragment (scFv) or an IgG4 based backbone. The difference in the size of the backbones translates into different half-lifes and kinetics. These different TM formats can be used to customize UniCAR therapy according to the given circumstances. We were able to purify and perform various experiments with the mentioned TMs. We could show binding to the target cells using flow cytometry. We could also show that the TMs could be used to induce specific lysis in vitro on monolayer cells and spheroids. We were also able to show UniCAR T cell infiltration into the spheroids and their consequent activation. Moreover, in mouse models the TMs could be used for P

Immunotherapy based on chimeric antigen receptor (CAR) T-cells has demonstrated remarkable therapeutic effects, particularly against some hematological cancers. A versatile adaptor CAR system called RevCAR, consisting of RevCAR T-cells and a bispecific target module (RevTM) has been developed to overcome severe side effects associated with conventional CAR T-cell therapy. As the activity of RevCAR T-cells can be steered based on the availability of RevTM, working as an on/off switch, the system can be immediately turned off if side effects occur. Furthermore, the RevCAR system is highly flexible, since the same RevCAR T-cell can be directed towards different tumor-associated antigens (TAA) simply by adding RevTMs with different specificities. However, the effectiveness of CAR T-cells against solid tumors remains limited particularly due to their immunosuppressive tumor microenvironment (TME). To overcome these hurdles, we have established a novel RevCAR system targeting immune checkpoint molecules such as PD-L1, which are frequently overexpressed by cancer cells to suppress immune responses. We have constructed novel RevTMs that can redirect RevCAR T-cells to kill tumor cells that express such immune checkpoint molecules. Furthermore, true AND gate tumor targeting was achieved by targeting a TAA in addition to PD-L1 in a combinatorial manner using our Dual-RevCAR system. In this way, targeting PD-L1 not only results in Dual-RevCAR T-cell activation but simultaneously blocks the immunosuppressive PD-L1/PD-1 axis. Altogether, we have turned an immunosuppressive marker into an immune-activating signal that might modulate the TME in a beneficial manner, showing promise for the development of an effective immunotherapy against solid tumors.

We introduce the NASA/IPAC Extragalactic Database (NED) Local Volume Sample (NED-LVS), a subset of $\sim$1.9 million objects with distances out to 1000~Mpc. We use UV and IR fluxes available in NED from all-sky surveys to derive physical properties, and estimate the completeness relative to the expected local luminosity density. The completeness relative to NIR luminosities (which traces a galaxy's stellar mass) is roughly 100% at $D<$30~Mpc and remains moderate (70%) out to 300~Mpc. For brighter galaxies ($\gtrsim L^{*}$), NED-LVS is $\sim$100% complete out to $\sim$400~Mpc. When compared to other local Universe samples (GLADE and HECATE), all three are $\sim$100% complete below 30~Mpc. At distances beyond $\sim$80~Mpc, NED-LVS is more complete than both GLADE and HECATE by $\sim$10-20%. NED-LVS is the underlying sample for the NED gravitational wave follow-up (NED-GWF) service, which provides prioritized lists of host candidates for GW events within minutes of alerts issued by the LIGO-Virgo-KAGRA collaboration. We test the prioritization of galaxies in the volume of GW170817 by 3 physical properties, where we find that both stellar mass and inverse specific star formation rate place the correct host galaxy in the top ten. In addition, NED-LVS can be used for a wide variety of other astrophysical studies: galaxy evolution, star formation, large-scale structure, galaxy environments, and more. The data in NED are updated regularly, and NED-LVS will be updated concurrently. Consequently, NED-LVS will continue to provide an increasingly complete sample of galaxies for a multitude of astrophysical research areas for years to come., Comment: 46 pages, 16 Figures, 7 Tables, Accepted for publication in ApJS

Contains fulltext : 282534.pdf (Publisher’s version ) (Open Access), BACKGROUND AND PURPOSE: Cerebrospinal fluid (CSF) real-time quaking-induced conversion (RT-QuIC) has a high degree of sensitivity and specificity for the diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) and this has led to its being included in revised European CJD Surveillance Network diagnostic criteria for sCJD. As CSF RT-QuIC becomes more widely established, it is crucial that the analytical performance of individual laboratories is consistent. The aim of this ring-trial was to ascertain the degree of concordance between European countries undertaking CSF RT-QuIC. METHODS: Ten identical CSF samples, seven from probable or neuropathologically confirmed sCJD and three from non-CJD cases, were sent to 13 laboratories from 11 countries for RT-QuIC analysis. A range of instrumentation and different recombinant prion protein substrates were used. Each laboratory analysed the CSF samples blinded to the diagnosis and reported the results as positive or negative. RESULTS: All 13 laboratories correctly identified five of the seven sCJD cases and the remaining two sCJD cases were identified by 92% of laboratories. Of the two sCJD cases that were not identified by all laboratories, one had a disease duration >26 months with a negative 14-3-3, whilst the remaining case had a 4-month disease duration and a positive 14-3-3. A single false positive CSF RT-QuIC result was observed in this study. CONCLUSIONS: This study shows that CSF RT-QuIC demonstrates an excellent concordance between centres, even when using a variety of instrumentation, recombinant prion protein substrates and CSF volumes. The adoption of CSF RT-QuIC by all CJD surveillance centres is recommended.

Contains fulltext : 282534.pdf (Publisher’s version ) (Open Access), BACKGROUND AND PURPOSE: Cerebrospinal fluid (CSF) real-time quaking-induced conversion (RT-QuIC) has a high degree of sensitivity and specificity for the diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) and this has led to its being included in revised European CJD Surveillance Network diagnostic criteria for sCJD. As CSF RT-QuIC becomes more widely established, it is crucial that the analytical performance of individual laboratories is consistent. The aim of this ring-trial was to ascertain the degree of concordance between European countries undertaking CSF RT-QuIC. METHODS: Ten identical CSF samples, seven from probable or neuropathologically confirmed sCJD and three from non-CJD cases, were sent to 13 laboratories from 11 countries for RT-QuIC analysis. A range of instrumentation and different recombinant prion protein substrates were used. Each laboratory analysed the CSF samples blinded to the diagnosis and reported the results as positive or negative. RESULTS: All 13 laboratories correctly identified five of the seven sCJD cases and the remaining two sCJD cases were identified by 92% of laboratories. Of the two sCJD cases that were not identified by all laboratories, one had a disease duration >26 months with a negative 14-3-3, whilst the remaining case had a 4-month disease duration and a positive 14-3-3. A single false positive CSF RT-QuIC result was observed in this study. CONCLUSIONS: This study shows that CSF RT-QuIC demonstrates an excellent concordance between centres, even when using a variety of instrumentation, recombinant prion protein substrates and CSF volumes. The adoption of CSF RT-QuIC by all CJD surveillance centres is recommended.

Sustainability reports are regarded as important tools in offering information about the environmental, social, economic, and institutional performance of an institution, and in demonstrating a commitment to matters related to sustainable development. But even though sustainability reporting has been used by a variety of higher education institutions to date, it is not as widely practiced as it should be. To further investigate this topic, a twofold approach was used: a study focusing on sustainability reporting approaches deployed in a sample of 30 universities across a set of countries; and a survey with a sample of 72 universities from different global regions to assess the extent to which they are deploying sustainability reporting as part of their activities. The scientific value of the paper resides in the fact that it offers a comprehensive overview of the subject matter of sustainability reporting, and how higher education institutions handle it. It also outlines the efforts in developing these documents which may catalyse further progress in this key area.

Dendritic cells (DCs) play a key role in the orchestration of antitumor immunity. Activated DCs efficiently enhance antitumor effects mediated by natural killer cells and T lymphocytes. Conversely, tolerogenic DCs essentially contribute to an immunosuppressive tumor microenvironment. Thus, DCs can profoundly influence tumor progression and clinical outcome of tumor patients. To gain novel insights into the role of human DCs in pancreatic ductal adenocarcinoma (PDAC), we explored the frequency, spatial organization, and clinical significance of conventional DCs type 1 (cDC1s) and type 2 (cDC2s) and plasmacytoid DCs (pDCs) in primary PDAC tissues. A higher density of whole tumor area (WTA)- and tumor stroma (TS)-infiltrating cDC1s was significantly associated with better disease-free survival (DFS). In addition, an increased frequency of intraepithelial tumor-infiltrating cDC2s was linked to better DFS and overall survival (OS). Furthermore, an increased density of WTA- and TS-infiltrating pDCs tended to improve DFS. Moreover, a higher frequency of WTA- and TS-infiltrating cDC1s and pDCs emerged as an independent prognostic factor for better DFS and OS. These findings indicate that tumor-infiltrating DCs can significantly influence the clinical outcome of PDAC patients and may contribute to the design of novel treatment options that target PDAC-infiltrating DCs.

Sustainability reports are regarded as important tools in offering information about the environmental, social, economic, and institutional performance of an institution, and in demonstrating a commitment to matters related to sustainable development. But even though sustainability reporting has been used by a variety of higher education institutions to date, it is not as widely practiced as it should be. To further investigate this topic, a twofold approach was used: a study focusing on sustainability reporting approaches deployed in a sample of 30 universities across a set of countries; and a survey with a sample of 72 universities from different global regions to assess the extent to which they are deploying sustainability reporting as part of their activities. The scientific value of the paper resides in the fact that it offers a comprehensive overview of the subject matter of sustainability reporting, and how higher education institutions handle it. It also outlines the efforts in developing these documents which may catalyse further progress in this key area.

We present the first public release of ShapePipe, an open-source and modular weak-lensing measurement, analysis, and validation pipeline written in Python. We describe the design of the software and justify the choices made. We provide a brief description of all the modules currently available and summarise how the pipeline has been applied to real Ultraviolet Near-Infrared Optical Northern Survey data. Finally, we mention plans for future applications and development. The code and accompanying documentation are publicly available on GitHub., Comment: 7 pages, 2 figures, accepted for publication in A&A

A common library, TMDlib2, for Transverse-Momentum-Dependent distributions (TMDs) and unintegrated parton distributions (uPDFs) is described, which allows for easy access of commonly used TMDs and uPDFs, providing a three-dimensional (3D) picture of the partonic structure of hadrons. The tool TMDplotter allows for web-based plotting of distributions implemented in TMDlib2, together with collinear pdfs as available in LHAPDF., Comment: 18 pages, 3 figures. Updated with version published in EPJC

A common library, TMDlib2, for Transverse-Momentum-Dependent distributions (TMDs) and unintegrated parton distributions (uPDFs) is described, which allows for easy access of commonly used TMDs and uPDFs, providing a three-dimensional (3D) picture of the partonic structure of hadrons. The tool TMDplotter allows for web-based plotting of distributions implemented in TMDlib2, together with collinear pdfs as available in LHAPDF., Comment: 18 pages, 3 figures. Updated with version published in EPJC

Educational inequalities in all-cause mortality have been observed for decades. However, the underlying biological mechanisms are not well known. We aimed to assess the role of DNA methylation changes in blood captured by epigenetic clocks in explaining these inequalities. Data were from eight prospective population-based cohort studies, representing 13,021 participants. First, educational inequalities and their portion explained by Horvath DNAmAge, Hannum DNAmAge, DNAmPhenoAge, and DNAmGrimAge epigenetic clocks were assessed in each cohort via counterfactual-based mediation models, on both absolute (hazard difference) and relative (hazard ratio) scales, and by sex. Second, estimates from each cohort were pooled through a random effect meta-analysis model. Men with low education had an excess mortality from all causes of 57 deaths per 10,000 person-years (95% confidence interval (CI): 38, 76) compared to their more advantaged counterparts. For women, the excess mortality was 4 deaths per 10,000 person-years (95% CI: -11, 19). On the relative scale, educational inequalities corresponded to hazard ratios of 1.33 (95% CI: 1.12, 1.57) for men and 1.15 (95% CI: 0.96, 1.37) for women. DNAmGrimAge accounted for the largest proportion, approximately 50%, of the educational inequalities for men, while the proportion was negligible for women. Most of this mediation was explained by differential effects of unhealthy lifestyles and morbidities of the WHO risk factors for premature mortality. These results support DNA methylation-based epigenetic aging as a signature of educational inequalities in life expectancy emphasizing the need for policies to address the unequal social distribution of these WHO risk factors.

Cancer stem cells (CSCs) are pluripotent and highly tumorigenic cells that can re-populate a tumor and cause relapses even after initially successful therapy. Like tissue stem cells, CSCs possess enhanced DNA repair mechanisms. An active DNA damage response alleviates the increased oxidative and replicative stress and leads to therapy resistance. On the other hand, mutations in DNA repair genes cause genomic instability, thereby driving tumor evolution and developing highly aggressive CSC phenotypes. However, the role of DNA repair proteins in CSCs extends beyond the level of DNA damage. In recent years, more and more studies reported the unexpected role of DNA repair proteins in the regulation of transcription, CSC signaling pathways, intracellular levels of reactive oxygen species (ROS), and epithelial-mesenchymal transition (EMT). Moreover, DNA damage signaling plays an essential role in the immune response towards tumor cells. Due to its high importance for the CSC phenotype and treatment resistance, the DNA damage response is a promising target for individualized therapies. Furthermore, understanding the dependence of CSC on DNA repair pathways can be therapeutically exploited to induce synthetic lethality and sensitize CSCs to anti-cancer therapies. This review discusses the different roles of DNA repair proteins in CSC maintenance and their potential as therapeutic targets.

According to the literature, the autoantigen La is involved in Cap-independent translation. It was proposed that one prerequisite for this function is the formation of a protein dimer. However, structural analyses argue against La protein dimers. Noteworthy to mention, these structural analyses were performed under reducing conditions. Here we describe that La protein can undergo redox-dependent structural changes. The oxidized form of La protein can form dimers, oligomers and even polymers stabilized by disulfide bridges. The primary sequence of La protein contains three cysteine residues. Only after mutation of all three cysteine residues to alanine La protein becomes insensitive to oxidation, indicating that all three cysteines are involved in redox-dependent structural changes. Biophysical analyses of the secondary structure of La protein support the redox-dependent conformational changes. Moreover, we identified monoclonal anti-La antibodies (anti-La mAbs) that react with either the reduced or oxidized form of La protein. Differential reactivities to the reduced and oxidized form of La protein were also found in anti-La sera of autoimmune patients.

Already decades ago, we and many other groups showed a nucleo-cytoplasmic translocation of La protein in cultured cells. This shuttling of La protein was seen after e.g. UV irradiation, virus infections, hydrogen peroxide exposure, and Fenton reaction based on iron or copper ions. In common, all these conditions are somehow related to oxidative stress. Unfortunately, all these harsh conditions could also cause an artificial release of La protein. Even until today, the shut-tling and a cytoplasmic function of La/SS-B are controversially discussed. Moreover, the driving mechanism for the shuttling of La protein remains unclear. Recently we showed that La protein undergoes redox dependent conformational changes. Moreover, we developed anti-La mono-clonal antibodies (anti-La mAbs) which are specific for either the reduced form of La protein or the oxidized form. Using these tools, here we show that redox dependent conformational changes are the driving force for the shuttling of La protein. Moreover, we show that transloca-tion of La protein to the cytoplasm can be triggered in a ligand/receptor dependent manner un-der physiological conditions. We show that ligands of toll-like receptors lead to a redox de-pendent shuttling of La protein. The shuttling of La protein depends on the redox status of the respective cell type. Endothelial cells are usually resistant to the shuttling of La protein while dendritic cells are highly sensitive. However, deprivation of intracellular reducing agents in endothelial cells turns endothelial cells sensitive to a redox dependent shuttling of La protein.

Since the first description of the nuclear autoantigens in the late sixties and early seventies of the last century we and many other groups have tried with difficulty to establish monoclonal anti-bodies (mabs) against nuclear antigens including to the autoantigen La/SS-B. To date, only a few anti-La mabs have been derived by conventional hybridoma technology. However, these pre-viously described anti-La mabs are not bona fide autoantibodies as they recognize either human La specific- or cryptic or post-translationally modified epitopes which are not accessible on na-tive mouse La protein. Herein we present a series of novel murine anti-La mabs including truly autoreactive ones. These mabs were elicited from a human La transgenic animal through adop-tive transfer of T cells from non-transgenic mice immunized with human La antigen. Detailed epitope and paratope analyses experimentally confirm the hypothesis that somatic hypermuta-tions occurring during T cell dependent maturation can lead to autoreactivity to the nuclear au-toantigen La/SS-B.

The assessment of the exposure of aquatic wildlife to complex environmental mixtures of chemicals originating from both point and diffuse sources and evaluating the potential impact thereof constitutes a significant step towards mitigating toxic pressure and the improvement of ecological status. In the current proof-of-concept study, we demonstrate the potential of a novel Aggregated Biomarker Response approach involving a comprehensive set of biomarkers to identify complex exposure and impacts on wild brown trout (Salmo trutta fario). Our scenario used a small lowland river in Germany (Holtemme river in the Elbe river catchment) impacted by two wastewater treatment plants (WWTP) and diffuse agricultural runoff as a case study. The trout were collected along a pollution gradient (characterised in a parallel study) in the river. Compared to fish from the reference site upstream of the first WWTP, the trout collected downstream of the WWTPs showed a significant increase in micronucleus formation, phase I and II enzyme activities, and oxidative stress parameters in agreement with increasing exposure to various chemicals. By integrating single biomarker responses into an Aggregated Biomarker Response (ABR) approach, the two WWTPs' contribution to the observed toxicity could be clearly differentiated. The ABR results were supported by chemical analyses and whole transcriptome data, which revealed alterations of steroid biosynthesis and associated pathways, including an anti-androgenic effect, as some of the key drivers of the observed toxicity. Overall, this combined approach of in situ biomarker responses complemented with molecular pathway analysis allowed for a comprehensive ecotoxicological assessment of fish along the river. This study provides evidence for specific hazard potentials caused by mixtures of agricultural and WWTP derived chemicals at sublethal concentrations. Using aggregated biomarker responses combined with chemical analyses enabled an evidence-based

It has been observed in the literature that measurements of low-mass Drell-Yan (DY) transverse momentum spectra at low center-of-mass energies $\sqrt{s}$ are not well described by perturbative QCD calculations in collinear factorization in the region where transverse momenta are comparable with the DY mass. We examine this issue from the standpoint of the Parton Branching (PB) method, combining next-to-leading-order (NLO) calculations of the hard process with the evolution of transverse momentum dependent (TMD) parton distributions. We compare our predictions with experimental measurements at low DY mass, and find very good agreement.In addition we use the low mass DY measurements at low $\sqrt{s}$ to determine the width $q_s$ of the intrinsic Gauss distribution of the PB-TMDs at low evolution scales. We find values close to what has earlier been used in applications of PB -TMDs to high-energy processes at the Large Hadron Collider (LHC) and HERA. We find that at low DY mass and low $\sqrt{s}$ even in the region of $p_t/m_{DY} \sim 1$ the contribution of multiple soft gluon emissions (included in the PB-TMDs) is essential to describe themeasurements, while at larger masses ($m_{DY} \sim m_{Z}$) and LHC energies the contribution from soft gluons in the region of $p_t/m_{DY}\sim 1$ is small., Comment: v2: references, clarifications and discussion added; results unchanged Version to appear in EPJC

Magnesium, titanium, special steel and composites are, together with aluminium, important in light-weight construction materials. About two thirds of Mg metal is used in Mg and Al alloys. These light-weight materials are utilised in the car industry (c.40% of Mg) and the electronics industry amongst others. One third of Mg is used in the metal industry, particularly in the production of cast iron and thermal reduction of metals. China is the biggest producer of Mg, c.80-85%. The extraction and production of Mg in an environmentally-friendly and energy-saving process in China and Canada is discussed and the effect of the Corona-virus considered for Mg production in China and for global markets, and alternative strategies are suggested., Magnesium, titanium, special steel and composites are, together with aluminium, important in light-weight construction materials. About two thirds of Mg metal is used in Mg and Al alloys. These light-weight materials are utilised in the car industry (c.40% of Mg) and the electronics industry amongst others. One third of Mg is used in the metal industry, particularly in the production of cast iron and thermal reduction of metals. China is the biggest producer of Mg, c.80-85%. The extraction and production of Mg in an environmentally-friendly and energy-saving process in China and Canada is discussed and the effect of the Corona-virus considered for Mg production in China and for global markets, and alternative strategies are suggested.

Contains fulltext : 224915.pdf (publisher's version ) (Open Access)

Somitogenesis, the primary segmentation of the vertebrate embryo, is associated with oscillating genes that interact with a wave of cell differentiation. The necessity of cell-matrix adherence and embryonic tension, however, suggests that mechanical cues are also involved. To explicitly investigate this, we applied surplus axial strain to live chick embryos. Despite substantial deformations, the embryos developed normally and somite formation rate was unaffected. Surprisingly, however, we observed slow cellular reorganizations of the most elongated somites into two or more well-shaped daughter somites. In what appeared to be a regular process of boundary formation, somites divided and fibronectin was deposited in between. Cell counts and morphology indicated that cells from the somitocoel underwent mesenchymal-epithelial transition; this was supported by a Cellular Potts model of somite division. Thus, although somitogenesis appeared to be extremely robust, we observed new boundary formation in existing somites and conclude that mechanical strain can be morphologically instructive.Poultry Embryology; Mechanical Modeling; Developmental Biology

Background The importance and frequency of peripheral arterial occlusive disease (PAOD) are often underestimated. In recent years a link between PAOD and chronic kidney disease (CKD) has been more and more frequently described. In dialysis patients PAOD is the most frequent reason for an amputation. Many risk factors, which are already of great importance for the development of arteriosclerosis, also play a decisive role in the development of CKD. Patients who suffer from both PAOD and CKD have a higher mortality risk than patients with only one of these diseases. Objective The aim of the study was to investigate the association between the stage of CKD and the degree of PAOD as well as the investigation of further mutual risk factors. Material and methods A retrospective observational study was conducted in 168 patients with PAOD who underwent surgical treatment in the city hospital of Solingen between 2011 and 2014. Results A marked reduction of renal function (>= CKD 3) was found in 40% of the patients. The extent of CKD was significantly higher in advanced stages of PAOD and in addition to age was associated with a higher relative chance of having an advanced stage of PAOD (odds ratio [OR] 1.933, 97.5% confidence interval [CI] 1.236-3.092 and OR 1.065, 97.5% CI 1.018-1.118). Furthermore, the extent of hyperparathyroidism increased with higher PAOD stages (30% in PAOD stage IV); however, the severity of arterial hypertension and diabetes mellitus was independent of the PAOD stage. Conclusion The primary goal in patients suffering from PAOD should be to identify the presence of CKD and when possible to consequently treat it in addition to the classical risk factors, in order to be able to delay the progression of PAOD.