Radiation, risk perception and raw material exploration: science communication and social license

Poster presented at VMSG 2020. The book of abstracts for this meeting is available from VMSG via the link in this recordWithin our modern world, high-tech communications and low carbon energy and transport are growing the need for raw materials. Alkaline rocks and carbonatites are key sources for such raw materials. However, most alkaline rock and carbonatite-related ore deposits contain hundreds of parts per million thorium and smaller amounts of uranium. Therefore, radiation can be a key hazard at exploration and mine sites. Our study of publically available data on radiation levels at exploration projects and mines, compared with records of protests associated with these sites, shows that: - Radiation levels in ore deposits, waste materials and by-products from mines vary considerably between different sites. - Protests about exploration projects and mines are associated with a wide range of concerns, sometimes, but not always, including radiation. - In some cases high levels of radiation-related concern are found at sites with lower radiation levels. - Insufficient trust and community engagement are repeatedly associated with protests associated with exploration and mine sites. Perception of radiation risk is important and needs to be addressed at all exploration projects in alkaline rocks and carbonatites. Background levels of radiation and concentrations of uranium and thorium in ore should be published as early as possible in exploration, even if these levels are low. No information is likely to lead to assumption of a problem. More broadly, best practices for good community – company relations and communication echo the best practice lessons learned in volcanic and other risk management scenarios and reflect the wider issues about science engagement and communication. Trust development, community engagement and transparency are essential in all cases and are key to effective relationships between communities and specialists working together in many circumstances. Acknowledgement: This project has received funding from the European Union's Horizon 2020 research and innovation programme (grant agreement no. 689909).European Union Horizon 202

Increased receptor affinity of SARS-CoV-2: a new immune escape mechanism.

‘Affinity escape’: Novel SARS-CoV-2 variants may escape immunity by raising the RBD-ACE2 affinity high enough to outcompete the avidity of neutralizing antibodies

Ion rocket system research and development Final report, 24 Feb. 1964 - 25 Jun. 1965

Design studies and testing of ion rocket engine and zero gravity feed syste

Molecular definition of severe acute respiratory syndrome coronavirus 2 receptor-binding domain mutations: Receptor affinity versus neutralization of receptor interaction.

BACKGROUND Several new variants of SARS-CoV-2 have emerged since fall 2020 which have multiple mutations in the receptor-binding domain (RBD) of the spike protein. It is unclear which mutations affect receptor affinity versus immune recognition. METHODS We produced wild type RBD, RBD with single mutations (E484K, K417N, or N501Y) or with all three mutations combined and tested their binding to ACE2 by biolayer interferometry (BLI). The ability of convalescent sera to recognize RBDs and block their interaction with ACE2 was tested as well. RESULTS We demonstrated that single mutation N501Y increased binding affinity to ACE2 but did not strongly affect its recognition by convalescent sera. In contrast, single mutation E484K had almost no impact on the binding kinetics, but essentially abolished recognition of RBD by convalescent sera. Interestingly, combining mutations E484K, K417N, and N501Y resulted in a RBD with both features: enhanced receptor binding and abolished immune recognition. CONCLUSIONS Our data demonstrate that single mutations either affect receptor affinity or immune recognition while triple mutant RBDs combine both features

Challenges in geological scientific fieldwork at the Kaiserstuhl carbonatite, Germany

This is the final version. Available from the European Commission via the link in this recordHiTech AlkCarb (New geomodels to explore deeper for High-Technology critical raw materials in Alkaline rocks and Carbonatites) project deliverable D6.3This report describes the challenges experienced during the geological scientific fieldwork at the Kaiserstuhl, Germany. Although the general perception of the public regarding the project is positive, the Municipality of Vogtsburg would have allowed the proposed ca. 400 m deep core drilling only if the HiTech AlkCarb project had taken an insurance policy for any potential environmental and social damage for the longest possible period after the drilling. This caution was the result of damage to buildings in the local town of Staufen im Breisgau and induced seismicity near Basel during geothermal energy projects. As it was not possible to obtain such an insurance, the drilling programme had to be abandoned. A company with an official exploration licence would have had the right to drill, although the lack of social licence for drilling may make an exploration licence hard to obtain in the region at the moment. Moreover, if the drilling programme had been contracted by a German governmental institution, such as a university or the geological survey, the drilling could have been carried out, as governmental institutions in Germany are automatically insured through the state. However, the local university HiTech AlkCarb project partner declined to take over running the drilling contract, and this probably reflects the perceived reputational risk of taking part in any drilling in the region at the moment. It is a good lesson in how damage to social licence can affect permitting. The recommendations to the European Commission arising from this experience are: • To assign any drilling in research and innovation projects, in Germany, and probably elsewhere in Europe, to a Government organisation where possible. • To publicise good practice examples and subsurface research projects (such as UK GEOS and similar) that can help to regain public confidence.European Union Horizon 202

Social licence for exploration/mining in Europe is influenced by other georesource projects such as deep and shallow geothermal energy

This is the final version.The Horizons 2020 HiTech AlkCarb project planned a 300m deep research drill hole, together with geological and geophysical explora<on studies, in southwest Germany, all carried out to international minerals exploration best practice. The geology and geophysics were well received by the local public, authori<es and poli<cians. However, the local politicians expressed concerns about the drill hole and requested a long term insurance policy against any nega<ve impacts. In prac<ce, this meant that the research team could not drill the hole. The cause of this concern was the nega<ve public percep<on of drilling caused by problems with two geothermal projects in the region. Although the HiTech AlkCarb team had been me<culous to apply mining-related best-prac<ce to public and official interac<ons, they had not realised the depth of feeling and loss of trust caused by the geothermal projects – in effect, the social licence for minerals explora<on drilling had been lost, and could not be regained during this project. The lesson here for expansion of mining or geothermal energy in Europe is clear – that in terms of social licence, the implica<ons of past and present geology-related projects must be considered together. The public does not discriminate and neither can industry, regional authori<es nor na<onal and interna<onal legislators.European Commissio

In situ delivery of nanoparticles formulated with micron-sized crystals protects from murine melanoma.

INTRODUCTION Intratumoral injections of novel therapeutics can activate tumor antigen-specific T cells for locoregional tumor control and may even induce durable systemic protection (against distant metastases) via recirculating T cells. Here we explored the possibility of a universal immunotherapy that promotes T-cell responses in situ and beyond, upon intratumoral injection of nanoparticles formulated with micron-sized crystals. METHODS Cucumber mosaic virus-like particles containing a tetanus toxin peptide (CuMVTT) were formulated with microcrystalline tyrosine (MCT) adjuvant and injected directly in B16F10 melanoma tumors. To further enhance immunogenicity, we loaded the nanoparticles with a TLR7/8 ligand and incorporated a universal tetanus toxin T-helper cell peptide. We assessed therapeutic efficacy and induction of local and systemic immune responses, including RNA sequencing, providing broad insight into the tumor microenvironment and correlates of protection. RESULTS MCT crystals were successfully decorated with CuMVTT nanoparticles. This 'immune-enhancer' formed immunogenic depots in injected tumors, enhanced polyfunctional CD8+ and CD4+ T cells, and inhibited B16F10 tumor growth locally and systemically. Local inflammation and immune responses were associated with upregulation of genes involved in complement activation and collagen formation. CONCLUSIONS Our new immune-enhancer turned immunologically cold tumors into hot ones and inhibited local and distant tumor growth. This type of immunotherapy does not require the identification of (patient-individual) relevant tumor antigens. It is well tolerated, non-infectious, and affordable, and can readily be upscaled for future clinical testing and broad application in melanoma and likely other solid tumors

DNA amplifications at 20q13 and MDM2 define distinct subsets of evolved breast and ovarian tumours.

DNA amplification seems to be particularly frequent in human breast tumours and has been associated with cancer evolution and aggressiveness. Recent data indicate that new events should be added to the list, such as the amplifications at chromosome 20q13 or the MDM2 gene. The present work aimed at determining the incidence and clinicopathological signification of these amplifications in a large series of breast and ovarian tumours. We tested 1371 breast and 179 ovarian tumours by Southern blotting and observed amplification of 20q13 in 5.4% breast and 2.8% ovarian carcinomas, whereas MDM2 was found amplified in 5.3% and 3.8% of breast and ovarian tumours respectively. MDM2 RNA expression levels were analysed in a subset of 57 breast tumours and overexpression was observed in 4/57 (7%) of the tumours. Elevated expression levels coincided with amplification of the gene. In breast cancer, 20q13 and MDM2 amplifications seem to define subsets of aggressive tumours. Indeed, 20q13 was correlated to axillary nodal involvement and occurred preferentially in younger patients (< 50 years). Furthermore, 20q13 correlated, as did MDM2 amplification, to aneuploidy. In parallel, we had also tested our tumour DNAs for amplification of CCND1, ERBB-2 and MYC, which made it possible to test for correlations with 20q13 or MDM2 amplifications. Whereas 20q13 showed a very strong correlation to CCND1 amplification, that of MDM2 was prevalent in MYC-amplified tumours. Interestingly, 20q13 and MDM2 amplifications showed some degree of correlation to each other, which may possibly be owing to the fact that both events occurred preferentially in aneuploid tumours. In ovarian cancer, no statistically significant correlation was observed. However, 20q13 amplification occurred preferentially in stage 3 tumours and MDM2 was correlated to ERBB-2 amplification. This may suggest that in ovarian tumours also, 20q13 and MDM2 amplifications occur in late or aggressive cancers

MART-1 peptide vaccination plus IMP321 (LAG-3Ig fusion protein) in patients receiving autologous PBMCs after lymphodepletion: results of a Phase I trial.

BACKGROUND: Immunotherapy offers a promising novel approach for the treatment of cancer and both adoptive T-cell transfer and immune modulation lead to regression of advanced melanoma. However, the potential synergy between these two strategies remains unclear. METHODS: We investigated in 12 patients with advanced stage IV melanoma the effect of multiple MART-1 analog peptide vaccinations with (n = 6) or without (n = 6) IMP321 (LAG-3Ig fusion protein) as an adjuvant in combination with lymphodepleting chemotherapy and adoptive transfer of autologous PBMCs at day (D) 0 (Trial registration No: NCT00324623). All patients were selected on the basis of ex vivo detectable MART-1-specific CD8 T-cell responses and immunized at D0, 8, 15, 22, 28, 52, and 74 post-reinfusion. RESULTS: After immunization, a significant expansion of MART-1-specific CD8 T cells was measured in 83% (n = 5/6) and 17% (n = 1/6) of patients from the IMP321 and control groups, respectively (P &lt; 0.02). Compared to the control group, the mean fold increase of MART-1-specific CD8 T cells in the IMP321 group was respectively &gt;2-, &gt;4- and &gt;6-fold higher at D15, D30 and D60 (P &lt; 0.02). Long-lasting MART-1-specific CD8 T-cell responses were significantly associated with IMP321 (P &lt; 0.02). At the peak of the response, MART-1-specific CD8 T cells contained higher proportions of effector (CCR7⁻ CD45RA⁺/⁻) cells in the IMP321 group (P &lt; 0.02) and showed no sign of exhaustion (i.e. were mostly PD1⁻CD160⁻TIM3⁻LAG3⁻2B4⁺/⁻). Moreover, IMP321 was associated with a significantly reduced expansion of regulatory T cells (P &lt; 0.04); consistently, we observed a negative correlation between the relative expansion of MART-1-specific CD8 T cells and of regulatory T cells. Finally, although there were no confirmed responses as per RECIST criteria, a transient, 30-day partial response was observed in a patient from the IMP321 group. CONCLUSIONS: Vaccination with IMP321 as an adjuvant in combination with lymphodepleting chemotherapy and adoptive transfer of autologous PBMCs induced more robust and durable cellular antitumor immune responses, supporting further development of IMP321 as an adjuvant for future immunotherapeutic strategies

Increased Receptor Affinity and Reduced Recognition by Specific Antibodies Contribute to Immune Escape of SARS-CoV-2 Variant Omicron.

In this report, we mechanistically reveal how the Variant of Concern (VOC) SARS-CoV-2 Omicron (B.1.1.529) escapes neutralizing antibody responses, by physio-chemical characterization of this variant in comparison to the wild-type Wuhan and the Delta variant (B.1.617.2). Convalescent sera, as well as sera obtained from participants who received two or three doses of mRNA vaccines (Moderna-mRNA-1273® or Pfizer-BNT162b2®), were used for comparison in this study. Our data demonstrate that both Delta, as well as Omicron variants, exhibit a higher affinity for the receptor ACE2, facilitating infection and causing antibody escape by receptor affinity (affinity escape), due to the reduced ability of antibodies to compete with RBD-receptor interaction and virus neutralization. In contrast, only Omicron but not the Delta variant escaped antibody recognition, most likely because only Omicron exhibits the mutation at E484A, a position associated with reduced recognition, resulting in further reduced neutralization (specificity escape). Nevertheless, the immunizations with RNA-based vaccines resulted in marked viral neutralization in vitro for all strains, compatible with the fact that Omicron is still largely susceptible to vaccination-induced antibodies, despite affinity- and specificity escape