A major update to the International Nuclear Workers Study was undertaken that allows us to report updated estimates of associations between radiation and site-specific solid cancer mortality. A cohort of 309 932 nuclear workers employed in France, the United Kingdom, and the United States were monitored for external radiation exposure. Associations of radiation with cancer mortality were quantified as the excess relative rate (ERR) per gray (Gy) using a maximum likelihood and a Markov chain Monte Carlo method (to stabilize estimates via a hierarchical regression). The analysis included 28 089 deaths due to solid cancer, the most common being lung, prostate, and colon cancer. Using maximum likelihood, positive estimates of ERR per Gy were obtained for stomach, colon, rectum, pancreas, peritoneum, larynx, lung, pleura/mesothelioma, bone and connective tissue, skin, prostate, testis, bladder, kidney, thyroid, and residual cancers. Negative estimates of ERR per Gy were found cancers of oral cavity and pharynx, esophagus, and ovary. A hierarchical model stabilized site-specific estimates of association, including for lung (ERR per Gy = 0.65; 95% credible interval [CrI], 0.24-1.07), prostate (ERR per Gy = 0.44; 95% CrI, -0.06 to 0.91), and colon cancer (ERR per Gy = 0.53; 95% CrI, -0.07 to 1.11). The results contribute evidence regarding associations between low-dose radiation and cancer.

OBJECTIVES:Understanding of long-term lung cancer risks from radon decay products (RDP) exposure derives largely from studies of uranium miners. We aimed to compare mortality for lung and other cancers to the general population, to estimate excess absolute rate (EAR) and excess relative rate (ERR) from RDP exposure, and to estimate the joint effects of RDP and cigarette smoking in extended follow-up of a cohort of 4137 male uranium miners from the US Colorado Plateau. METHODS:We extended mortality follow-up through 2016 and re-evaluated RDP exposure against original work history and mine records. We calculated standardised mortality ratios (SMRs) compared with a regional population, evaluated EAR of lung cancer mortality using standardised rate ratios and modelled ERR using Cox proportional hazards regression. We evaluated interactions of RDP with smoking pack-years, attained age (AA) and time-since-exposure (TSE). RESULTS:There were 695 lung cancer deaths, including 146 among never-smokers and light smokers. The overall SMR was >4; the EAR per unit RDP exposure increased substantially with smoking pack-years and decades of follow-up. Lung cancer ERR decreased with AA and TSE. ERR attenuation at high exposure rates was smaller than observed elsewhere. Joint effects of RDP and smoking were submultiplicative but greater-than-additive, appearing closer to multiplicative at lower RDP exposures. Pancreas was the only other site showing a significantly positive ERR per unit exposure. CONCLUSIONS:Excess rates of lung cancer mortality persist throughout the lifespan among this cohort of uranium miners. Information about RDP-smoking interactions is of interest for occupational and general population exposure.

Radon is a known cause of lung cancer. Protective standards for radon exposure are derived largely from studies of working populations that are prone to healthy worker survivor bias. This bias can lead to underprotection of workers and is a key barrier to understanding health effects of many exposures. We apply inverse probability weighting to study a set of hypothetical exposure limits among 4137 male, White and American Indian radon-exposed uranium miners in the Colorado Plateau followed from 1950 to 2005. We estimate cumulative risk of lung cancer through age 90 under hypothetical occupational limits. We estimate that earlier implementation of the current US Mining Safety and Health Administration annual standard of 4 working level months (implemented here as a monthly exposure limit) could have reduced lung cancer mortality from 16 of 100 workers to 6 of 100 workers (95% confidence interval, 3/100, 8/100), in contrast with previous estimates of 10 of 100 workers. Our estimate is similar to that among contemporaneous occupational cohorts. Inverse probability weighting is a simple and computationally efficient way address healthy worker survivor bias to contrast health effects of exposure limits and estimate the number of excess health outcomes under exposure limits at work.

A recent commentary on epidemiological studies of nuclear workers notes that these studies can provide radiation risk estimates that complement those derived from the study of Japanese atomic bomb survivors. The author asserts that the results from some nuclear worker studies are difficult to interpret due to the fact that ERR/Gy estimates vary across subcohorts, and subcohort-specific estimates are not always equal to estimates obtained in the overall study population. We discuss settings in which it is reasonable to expect that an estimate of association in a subcohort should be similar to an estimate obtained in the full cohort and settings in which a subcohort analysis may differ from the estimate obtained in a full cohort analysis. Focusing on the INWORKS study, we describe some of the steps taken to understand variation in estimates of ERR/Gy between subgroups and upon restrictions, as well as interpretation of estimates of external dose-mortality associations in the total study population.

Lateral organization of membrane proteins in the plasma membrane has been demonstrated to be essential for a vast number of cellular processes. Due to the short-lived nature of membrane domains, it is difficult to see protein clustering in vivo, and controlled in vitro macroscale induction of lipid domains in model membranes frequently necessitates the inclusion of clustering agents or unphysiological observation conditions. To overcome these difficulties, we have created a novel approach that combines optical trapping, thermoplasmonic-mediated membrane fusion, and confocal imaging to transport native plasma membrane proteins into phase-separated model systems. Fusion of a plasma membrane vesicle with a model membrane ensures the preservation of the correct, native protein orientation in the hybrid membrane, while simultaneously allowing us to monitor lateral protein segregation in real time. By examining the phase affinities of the influenza virus envelope proteins NA, HA, and M2 at physiologically relevant temperatures, we show the adaptability and potential of this method. Our generic platform can be utilized to look at the lateral arrangement of any fluorescently labeled plasma membrane component in a specific model membrane. Furthermore, it enables the initiation of chemical reactions or ligand interactions with native membrane proteins, opening up novel pathways for studying the dynamics of soft matter systems.

Supplementing influenza vaccines with additional protective antigens such as neuraminidase (NA) is a promising strategy for increasing the breadth of the immune response. Here, we improved the immunogenicity and stability of secreted recombinant NA (rNA) tetramers by covalently conjugating them onto the surface of AP205 capsid virus-like particles (cVLPs) using a Tag/Catcher ligation system. cVLP display increased the induction of IgG2a subclass anti-NA antibodies, which exhibited cross-reactivity with an antigenically distant homologous NA. It also reduced the single dose rNA amounts needed for protection against viral challenge in mice, demonstrating a dose-sparing effect. Moreover, effective cVLP-display was achieved across different NA subtypes, even when the conjugation was performed shortly before administration. Notably, the rNA-cVLP immunogenicity was retained upon mixing or co-administering with commercial vaccines. These results highlight the potential of this approach for bolstering the protective immune responses elicited by influenza vaccines.

The International Nuclear Workers Study (INWORKS) contributes knowledge on the dose-response association between predominantly low dose, low dose rate occupational exposures to penetrating forms of ionizing radiation and cause-specific mortality. By extending follow-up of 309,932 radiation workers from France (1968–2014), the United Kingdom (1955–2012), and the United States (1944–2016) we increased support for analyses of temporal variation in radiation-cancer mortality associations. Here, we examine whether age at exposure, time since exposure, or attained age separately modify associations between radiation and mortality from all solid cancers, solid cancers excluding lung cancer, lung cancer, and lymphohematopoietic cancers. Multivariable Poisson regression was used to fit general relative rate models that describe modification of the linear excess relative rate per unit organ absorbed dose. Given indication of greater risk per unit dose for solid cancer mortality among workers hired in more recent calendar years, sensitivity analyses considering the impact of year of hire on results were performed. Findings were reasonably compatible with those from previous pooled and country-specific analyses within INWORKS showing temporal patterns of effect measure modification that varied among cancers, with evidence of persistent radiation-associated excess cancer risk decades after exposure, although statistically significant temporal modification of the radiation effect was not observed. Analyses stratified by hire period (< 1958, 1958+) showed temporal patterns that varied; however, these analyses did not suggest that this was due to differences in distribution of these effect measure modifiers by hire year.

Sialosides containing C8-modified sialic acids are challenging synthetic targets but potentially useful probes for diagnostic substrate profiling of sialidases and elucidating the binding specificity of sialic acid-interacting proteins. Here, we demonstrate efficient chemoenzymatic methods for synthesizing para -nitrophenol-tagged α2–3- and α2–6-linked sialyl galactosides containing C8-acetamido, C8-azido, or C8-amino derivatized N -acetylneuraminic acid (Neu5Ac). High-throughput substrate specificity studies showed that the C8-modification of sialic acid significantly changes its recognition by sialidases from humans, various bacteria, and different influenza A and B viruses. Sialosides carrying Neu5Ac with a C8-azido modification were generally well tolerated by all the sialidases we tested, whereas sialosides containing C8-acetamido-modified Neu5Ac were only cleaved by selective bacterial sialidases. In contrast, sialosides with C8-amino-modified Neu5Ac were cleaved by a combination of selective bacterial and influenza A virus sialidases. These results indicate that sialosides terminated with a C8-amino or C8-acetamido-modified sialic acid can be used with other sialosides for diagnostic profiling of disease-causing sialidase-producing pathogens.

Background A major update to the International Nuclear Workers Study (INWORKS) was undertaken to strengthen understanding of associations between low-dose exposure to penetrating forms of ionising radiation and mortality. Here, we report on associations between radiation dose and mortality due to haematological malignancies. Methods We assembled a cohort of 309 932 radiation-monitored workers (269 487 [87%] males and 40 445 [13%] females) employed for at least 1 year by a nuclear facility in France (60 697 workers), the UK (147 872 workers), and the USA (101363 workers). Workers were individually monitored for external radiation exposure and followed-up from Jan 1, 1944, to Dec 31, 2016, accruing 1072 million person-years of follow-up. Radiation-mortality associations were quantified in terms of the excess relative rate (ERR) per Gy of radiation dose to red bone marrow for leukaemia excluding chronic lymphocytic leukaemia (CLL), as well as subtypes ofleukaemia, myelodysplastic syndromes, non-Hodgkin and Hodgkin lymphomas, and multiple myeloma. Estimates of association were obtained using Poisson regression methods. Findings The association between cumulative dose to red bone marrow, lagged 2 years, and leukaemia (excluding CLL) mortality was well described by a linear model (ERR per Gy 268, 90% CI 113 to 455, n=771) and was not modified by neutron exposure, internal contamination monitoring status, or period of hire. Positive associations were also observed for chronic myeloid leukaemia (957, 400 to 1791, n=122) and myelodysplastic syndromes alone (319, 035 to 733, n=163) or combined with acute myeloid leukaemia (155, 005 to 342, n=598). No significant association was observed for acute lymphoblastic leukaemia (425, -419 to 1932, n=49) or CLL (020, -181 to 221, n=242). A positive association was observed between radiation dose and multiple myeloma (162, 006 to 364, n=527) whereas minimal evidence of association was observed between radiation dose and non-Hodgkin lymphoma (027, -061 to 139, n=1146) or Hodgkin lymphoma (060, -364 to 483, n=122) mortality. Interpretation This study reports a positive association between protracted low dose exposure to ionising radiation and mortality due to some haematological malignancies. Given the relatively low doses typically accrued by workers in this study (16 mGy average cumulative red bone marrow dose) the radiation attributable absolute risk of leukaemia mortality in this population is low (one excess death in 10 000 workers over a 35-year period). These results can inform radiation protection standards and will provide input for discussions on the radiation protection system. Funding National Cancer Institute, Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health, Institut de Radioprotection et de S & ucirc;ret & eacute; Nucl & eacute;aire, Orano, Electricit & eacute; de France, UK Health Security Agency. Copyright (c) 2024. World Health Organization. Published by Elsevier Ltd/Inc/BV. All rights reserved.

In this article we review the history of key epidemiologicalstudies of populations exposed to ionizing radiation. We high-light historical and recent findings regarding radiation-associated risks for incidence and mortality of cancer andnon-cancer outcomes with emphasis on study design andmethods of exposure assessment and dose estimation alongwith brief consideration of sources of bias for a few of themore important studies. We examine the findings from theepidemiological studies of the Japanese atomic bomb survivors,persons exposed to radiation for diagnostic or therapeutic pur-poses, those exposed to environmental sources includingChornobyl and other reactor accidents, and occupationallyexposed cohorts. We also summarize results of pooled studies.These summaries are necessarily brief, but we provide refer-ences to more detailed information. We discuss possible futuredirections of study, to include assessment of susceptible popula-tions, and possible new populations, data sources, study designsand methods of analysis. @2024 by Radiation Research Society

Introduction Studies of nuclear workers provide insights into the health effects of ionizing radiation at levels relevant to contemporary workers and the general public. We evaluated the association between penetrating ionizing radiation exposure and cancer mortality subtypes in a large pooled cohort of US nuclear workers. Follow-up was extended an additional decade to improve power and examine cancers with longer latency. Materials and Methods The pooled cohort includes 101,363 workers from five US Department of Energy and Department of Defense nuclear facilities, followed for causes of death between 1944 and 2016. Workers were individually monitored for ionizing radiation exposure with the use of personal dosimeter badges. The association between cumulative external penetrating ionizing radiation exposure and cancer subtypes were modeled as the excess relative rate per Sievert (ERR Sv-1) using Cox regression. Results There were 13,568 cancer deaths during follow-up. We observed positive associations between ionizing radiation exposure and all solid cancer mortality (ERR Sv-1=0.19; 95%CI: -0.10, 0.52), and all lymphatic and hematopoietic cancers (ERR Sv-1=2.10; 95%CI: 0.97, 3.48). These associations were stronger among a contemporary sub cohort of workers first hired 1960 or later for both solid cancer (ERR Sv-1= 2.23; 95% CI: 1.13, 3.49) and all lymphatic and hematopoietic cancers (ERR Sv-1= 6.26; 95%CI: 2.86, 10.83). Additionally, we observed positive associations for several site specific lymphatic and hematopoietic cancer types, as well as lung cancer. In some instances, we observed modification by time since exposure and age at exposure. Conclusions This analysis confirms the association between low dose, low dose-rate radiation and leukemias, and strengthens the evidence base supporting the radiogenic nature of some solid cancers. The extended follow-up, individual dosimetry, and precise estimates provided by this large pooled analysis can better inform current radiological protection models.

ABSTRACT Most seasonal influenza vaccines are produced using hemagglutinin (HA) surface antigens from inactivated virions. However, virions are thought to be a suboptimal source for the less abundant NA surface antigen, which is also protective against severe disease. Here, we demonstrate that inactivated influenza virions are compatible with two modern approaches for improving protective NA antibody responses. Using a DBA/2J mouse model, we confirmed that the strong infection-induced NA inhibitory (NAI) antibody responses are only achieved by high dose immunizations of inactivated virions, likely due to low viral NA content. Based on this observation, we first produced virions with higher NA content by using reverse genetics to exchange the viral internal gene segments. Single immunizations with these inactivated virions enhanced NAI antibody responses, improved NA-based protection from a lethal viral challenge and allowed the development of natural immunity to the heterotypic challenge virus HA. Second, we combined inactivated virions with recombinant NA proteins. These mixtures increased NA-based protection following viral challenge and elicited stronger NA antibody responses than either component alone, especially when the NAs were homologous. Together, these results indicate that viral- and protein-based vaccines can be combined in a single syringe to improve protective antibody responses to influenza antigens.

Recombinant protein-based approaches are ideally suited for producing vaccine antigens that are not overly abundant in viruses, such as influenza neuraminidase (NA). However, obtaining sufficient quantities of recombinant viral surface antigens remains challenging, often resulting in the use of chimeric proteins with affinity tags that can invariably impact the antigen's properties. Here, we developed multistep chromatography approaches for purifying secreted recombinant NA (rNA) antigens that are derived from recent H1N1 and H3N2 viruses and produced using insect cells. Analytical analyses showed that these isolation procedures yielded homogenous tetrameric rNA preparations with consistent specific activities that were not possible from a common immobilized metal affinity chromatography purification procedure. The use of classical chromatography improved the rNA tetramer homogeneity by removing the requirement of the N-terminal poly-histidine affinity tag that was shown to promote higher order rNA oligomer formation. In addition, these procedures reduced the specific activity variation by eliminating the exposure to Ni2+ ions and imidazole, with the latter showing pH and NA subtype dependent effects. Together, these results demonstrate that purification by multistep chromatography improves the homogeneity of secreted rNAs and eliminates the need for affinity tag-based approaches that can potentially alter the properties of these recombinant antigens.

Introduction Ionizing radiation is an established carcinogen. Previously, the International Nuclear Workers Study (INWORKS), a pooled cohort of 308,297 workers from the United States (US), the United Kingdom (UK), and France followed 1944–2005, yielded an estimated excess relative rate of 0.51 per Gy (90% CI = 0.23, 0.82) for cancer mortality (19,748 deaths). Prior analyses suggested potential for healthy worker survivor bias and modification by age-at-exposure and time-since-exposure. Material and Methods We reanalyzed INWORKS data using the parametric g-formula, which can address healthy worker survivor bias. We estimated impacts of 2 hypothetical interventions: 1) reduce historical standards of worker equivalent doses to 5 mSv/year; and, 2) exposure to constant levels of 20 mGy/year vs. 5 mGy/year while at work. This approach requires models for exposure, employment and cause-specific mortality. Mortality models allow variation in the radiation-mortality association with age-at-exposure and time-since-exposure. Results Conditional on prior exposure and covariates, employment in the prior year was inversely associated with mortality among French workers but positively associated in US and UK workers. Associations between dose and cancer mortality were largest for doses received after age 45, lagged 30+ years. Approximately 224 cancer deaths per 1,000 workers were expected by age 90 in the pooled cohort. Relative to historical exposure levels (10% of working-years > 5 mGy/year), we estimated that a hypothetical standard at 5 mGy/year would result in 6.9 (95% CI = -5.7, 19) fewer cancer deaths per 1,000 workers by age 90. A constant exposure at 5 mGy/year (relative to 20 mGy/year) would have resulted in 13 (95% CI = -1.1, 27) fewer deaths. Conclusion Our results suggest that confounding by employment status was present, suggesting healthy worker survivor bias. The importance of doses at older ages and long times-since-exposure suggest continued need to assess potential impacts of occupational ionizing radiation exposures.

Objective To evaluate the effect of protracted low dose, low dose rate exposure to ionising radiation on the risk of cancer. Design Multinational cohort study. Setting Cohorts of workers in the nuclear industry in France, the UK, and the US included in a major update to the International Nuclear Workers Study (INWORKS). Participants 309 932 workers with individual monitoring data for external exposure to ionising radiation and a total follow-up of 10.7 million person years. Main outcome measures Estimates of excess relative rate per gray (Gy) of radiation dose for mortality from cancer. Results The study included 103 553 deaths, of which 28 089 were due to solid cancers. The estimated rate of mortality due to solid cancer increased with cumulative dose by 52% (90% confidence interval 27% to 77%) per Gy, lagged by 10 years. Restricting the analysis to the low cumulative dose range (0-100 mGy) approximately doubled the estimate of association (and increased the width of its confidence interval), as did restricting the analysis to workers hired in the more recent years of operations when estimates of occupational external penetrating radiation dose were recorded more accurately. Exclusion of deaths from lung cancer and pleural cancer had a modest effect on the estimated magnitude of association, providing indirect evidence that the association was not substantially confounded by smoking or occupational exposure to asbestos. Conclusions This major update to INWORKS provides a direct estimate of the association between protracted low dose exposure to ionising radiation and solid cancer mortality based on some of the world’s most informative cohorts of radiation workers. The summary estimate of excess relative rate solid cancer mortality per Gy is larger than estimates currently informing radiation protection, and some evidence suggests a steeper slope for the dose-response association in the low dose range than over the full dose range. These results can help to strengthen radiation protection, especially for low dose exposures that are of primary interest in contemporary medical, occupational, and environmental settings.

Most seasonal influenza vaccines are produced using hemagglutinin (HA) surface antigens from inactivated virions. However, virions are thought to be a suboptimal source for the less abundant neuraminidase (NA) surface antigen, which is also protective against severe disease. Here, we demonstrate that inactivated influenza virions are compatible with two modern approaches for improving protective antibody responses against NA. Using a DBA/2J mouse model, we show that the strong infection-induced NA inhibitory (NAI) antibody responses are only achieved by high dose immunizations of inactivated virions, likely due to the low viral NA content. Based on this observation, we first produced virions with higher NA content by using reverse genetics to exchange the viral internal gene segments. Single immunizations with these inactivated virions showed enhanced NAI antibody responses and improved NA-based protection from a lethal viral challenge while also allowing for the development of natural immunity to the heterotypic challenge virus HA. Second, we combined inactivated virions with recombinant NA protein antigens. These combination vaccines increased NA-based protection following viral challenge and elicited stronger antibody responses against NA than either component alone, especially when the NAs possessed similar antigenicity. Together, these results indicate that inactivated virions are a flexible platform that can be easily combined with protein-based vaccines to improve protective antibody responses against influenza antigens.

Influenza A viruses and the bacterium Streptococcus pneumoniae (pneumococci) both express neuraminidases that catalyze release of sialic acid residues from oligosaccharides and glycoproteins. Although these respiratory pathogen neuraminidases function in a similar environment, it remains unclear if these enzymes use similar mechanisms for sialic acid cleavage. Here, we compared the enzymatic properties of neuraminidases from two influenza A subtypes (N1 and N2) and the pneumococcal strain TIGR4 (NanA, NanB, and NanC). Insect cell-produced N1 and N2 tetramers exhibited calciumdependent activities and stabilities that varied with pH. In contrast, E. coli-produced NanA, NanB, and NanC were isolated as calcium insensitive monomers with stabilities that were more resistant to pH changes. Using a synthetic substrate (MUNANA), all neuraminidases showed similar pH optimums (pH 6-7) that were primarily defined by changes in catalytic rate rather than substrate binding affinity. Upon using a multivalent substrate (fetuin sialoglycans), much higher specific activities were observed for pneumococcal neuraminidases that contain an additional lectin domain. In virions, N1 and fetuin that was lost upon the addition of detergent, indicating the sialic acid-binding capacity of neighboring hemagglutinin molecules likely contributes to catalysis of natural multivalent substrates. These results demonstrate that influenza and pneumococcal neuraminidases have evolved similar yet distinct strategies to optimize their catalytic activity.

BACKGROUND The risk of solid cancers from low-level protracted ionizing radiation is not well characterized. Nuclear workers provide valuable information on the effects of ionizing radiation in contemporary exposure scenarios relevant to workers and the public. METHODS We evaluated the association between penetrating ionizing radiation exposure and solid cancer mortality among a pooled cohort of nuclear workers in the USA, with extended follow-up to examine cancers with long latencies. This analysis includes 101 363 workers from five nuclear facilities, with 12 069 solid cancer deaths between 1944 and 2016. The association between cumulative equivalent dose measured in sieverts (Sv) and solid cancer subtypes were modelled as the excess relative rate per Sv (ERR Sv-1) using Cox regression. RESULTS For the association between ionizing radiation exposure and all solid cancer mortality we observed an elevated rate (ERR Sv-1=0.19; 95% CI: -0.10, 0.52), which was higher among a contemporary sub-cohort of workers first hired in 1960 or later (ERR Sv-1= 2.23; 95% CI: 1.13, 3.49). Similarly, we observed an elevated rate for lung cancer mortality (ERR Sv-1= 0.65; 95% CI: 0.09, 1.30) that was higher among contemporary hires (ERR Sv-1= 2.90; 95% CI: 1.00, 5.26). CONCLUSIONS Although concerns remain about confounding, measurement error and precision, this analysis strengthens the evidence base indicating there are radiogenic risks for several solid cancer types.