The unremarkable alveolar epithelial glycocalyx: a thorium dioxide-based electron microscopic comparison after heparinase or pneumolysin treatment.

Recent investigations analyzed in depth the biochemical and biophysical properties of the endothelial glycocalyx. In comparison, this complex cell-covering structure is largely understudied in alveolar epithelial cells. To better characterize the alveolar glycocalyx ultrastructure, unaffected versus injured human lung tissue explants and mouse lungs were analyzed by transmission electron microscopy. Lung tissue was treated with either heparinase (HEP), known to shed glycocalyx components, or pneumolysin (PLY), the exotoxin of Streptococcus pneumoniae not investigated for structural glycocalyx effects so far. Cationic colloidal thorium dioxide (cThO2) particles were used for glycocalyx glycosaminoglycan visualization. The level of cThO2 particles orthogonal to apical cell membranes (≙ stained glycosaminoglycan height) of alveolar epithelial type I (AEI) and type II (AEII) cells was stereologically measured. In addition, cThO2 particle density was studied by dual-axis electron tomography (≙ stained glycosaminoglycan density in three dimensions). For untreated samples, the average cThO2 particle level was ≈ 18 nm for human AEI, ≈ 17 nm for mouse AEI, ≈ 44 nm for human AEII and ≈ 35 nm for mouse AEII. Both treatments, HEP and PLY, resulted in a significant reduction of cThO2 particle levels on human and mouse AEI and AEII. Moreover, a HEP- and PLY-associated reduction in cThO2 particle density was observed. The present study provides quantitative data on the differential glycocalyx distribution on AEI and AEII based on cThO2 and demonstrates alveolar glycocalyx shedding in response to HEP or PLY resulting in a structural reduction in both glycosaminoglycan height and density. Future studies should elucidate the underlying alveolar epithelial cell type-specific distribution of glycocalyx subcomponents for better functional understanding.

Estimating the Risk for Secondary Cancer After Targeted α-Therapy with 211At Intraperitoneal Radioimmunotherapy.

Intraperitoneal 211At-based targeted α-therapy (TAT) may hold great promise as an adjuvant therapy after surgery and chemotherapy in epithelial ovarian cancer to eradicate any remaining undetectable disease. This implies that it will also be delivered to patients possibly already cured by the primary treatment. An estimate of long-term risks is therefore sought to determine whether the treatment is justified. Methods: Baseline data for risk estimates of α-particle irradiation were collected from published studies on excess cancer induction and mortality for subjects exposed to either 224Ra treatments or Thorotrast contrast agent (25% ThO2 colloid, containing 232Th). Organ dosimetry for 224Ra and Thorotrast irradiation were taken from the literature. These organ-specific risks were then applied to our previously reported dosimetry for intraperitoneal 211At-TAT patients. Results: Risk could be estimated for 10 different organ or organ groups. The calculated excess relative risk per gray (ERR/Gy) could be sorted into 2 groups. The lower-ERR/Gy group, ranging up to a value of approximately 5, included trachea, bronchus, and lung, at 0.52 (95% CI, 0.21-0.82); stomach, at 1.4 (95% CI, -5.0-7.9); lymphoid and hematopoietic system, at 2.17 (95% CI, 1.7-2.7); bone and articular cartilage, at 2.6 (95% CI, 2.0-3.3); breast, at 3.45 (95% CI, -10-17); and colon, at 4.5 (95% CI, -3.5-13). The higher-ERR/Gy group, ranging from approximately 10 to 15, included urinary bladder, at 10.1 (95% CI, 1.4-23); liver, at 14.2 (95% CI, 13-16); kidney, at 14.9 (95% CI, 3.9-26); and lip, oral cavity, and pharynx, at 15.20 (95% CI, 2.73-27.63). Applying a typical candidate patient (female, age 65 y) and correcting for the reference population mortality rate, the total estimated excess mortality for an intraperitoneal 211At-monoclonal antibody treatment amounted to 1.13 per 100 treated. More than half this excess originated from urinary bladder and kidney, 0.29 and 0.34, respectively. Depending on various adjustments in calculation and assumptions on competing risks, excess mortality could range from 0.11 to 1.84 per 100 treated. Conclusion: Published epidemiologic data on lifelong detriment after α-particle irradiation and its dosimetry allowed calculations to estimate the risk for secondary cancer after 211At-based intraperitoneal TAT. Measures to reduce dose to the urinary organs may further decrease the estimated relative low risk for secondary cancer from 211At-monoclonal antibody-based intraperitoneal TAT.

Long-term incidence in hepatocellular carcinoma and intrahepatic bile duct cancer in Denmark, Finland, Norway and Sweden, role of Thorotrast?

We analyzed long-term incidence trends in liver cancer (including hepatocellular carcinoma and intrahepatic cholangiocarcinoma) with an aim to interpret the changes in terms of known risk factors and hypothesize that historical exposure to Thorotrast, a radiographic contrast medium emitting alpha particles, has changed population rates. The NORDCAN database was used to collect cancer registry data from Denmark (DK), Finland (FI), Norway (NO) and Sweden (SE), which we used from 1953 (DK, FI and NO) and 1960 (SE) through 2019. Thorotrast, which caused a 100-fold risk of liver cancer was used in DK and SE, and probably also in FI between 1930 and 1950, but not in NO. The incidence trend for liver cancer showed a broad maximum at around 1980, most prominent and statistically significant in SE and DK men and women, and in all countries, a steadily increasing trend towards the end of follow-up. Incidence for NO was lower than for the other countries and the rates showed no peaking at around 1980. Birth cohort analysis identified a transient risk which could be dated to a period between 1930 and 1950 in countries other than NO. Considering a lag time between Thorotrast use and liver cancer appearance, the large incidence peak around 1980 in DK and DE was probably contributed by Thorotrast but considering the ecological nature of the findings, the association should be considered cautiously as hypothesis generating. The late increase in liver cancer risk is most likely lifestyle related and largely preventable.

Long-term incidence and survival trends in cancer of the gallbladder and extrahepatic bile ducts in Denmark, Finland, Norway and Sweden with etiological implications related to Thorotrast.

Cancers of the gallbladder and extrahepatic bile ducts (called here "GBC" because gallbladder cancer is the main component) are rare in Europe, including the Nordic countries. Their incidence has varied for unknown reasons and we hypothesize that Thorotrast, a previously used carcinogenic radiographic contrast medium, has contributed to the incidence trends. We obtained incidence and survival data from the NORDCAN database, which includes cancer registry data from Denmark (DK), Finland (FI), Norway (NO) and Sweden (SE), which are globally the oldest national cancer databases, starting from 1943 in DK, 1953 in FI and NO and 1960 in SE, and extending to 2016. The incidence trend for GBC showed a broad maximum around 1980 in men (close to 3/100 000) and women (4/100 000), except for NO, where this phenomenon was not seen. In 1955, FI and NO incidence rates were equal but FI rates peaked and later declined similar to DK and SE rates. By 2010, the incidence was similar in all Nordic countries, for both men and women, at close to 2.0/100 000. Birth cohort analysis showed strong effects for countries other than NO. Relative 1-year survival increased for men from 20% to about 50% and similarly for women although at a 5 percentage points lower level. Survival in NO was better than in other countries in the 1980s. Thorotrast, causing a high risk of GBC, was extensively used in the Nordic countries between 1930 and end of 1940s, with the exception of NO, where these was no documented use. These data suggest that Thorotrast influenced GBC epidemiology and probably worsened survival in certain periods.

Mechanism of thorium-nitrate and thorium-dioxide induced cytotoxicity in normal human lung epithelial cells (WI26): Role of oxidative stress, HSPs and DNA damage.

Inhalation represents the most prevalent route of exposure with Thorium-232 compounds (Th-nitrate/Th-dioxide)/Th-containing dust in real occupational scenario. The present study investigated the mechanism of Th response in normal human alveolar epithelial cells (WI26), exposed to Th-nitrate or colloidal Th-dioxide (1-100 µg/ml, 24-72 h). Assessment in terms of changes in cell morphology, cell proliferation (cell count), plasma membrane integrity (lactate dehydrogenase leakage) and mitochondrial metabolic activity (MTT reduction) showed that Th-dioxide was quantitatively more deleterious than Th-nitrate to WI26 cells. TEM and immunofluorescence analysis suggested that Th-dioxide followed a clathrin/caveolin-mediated endocytosis, however, membrane perforation/non-endocytosis seemed to be the mode of Th internalization in cells exposed to Th-nitrate. Th-estimation by ICP-MS showed significantly higher uptake of Th in cells treated with Th-dioxide than with Th-nitrate at a given concentration. Both Th-dioxide and nitrate were found to increase the level of reactive oxygen species, which seemed to be responsible for lipid peroxidation, alteration in mitochondrial membrane potential and DNA-damage. Amongst HSPs, the protein levels of HSP70 and HSP90 were affected differentially by Th-nitrate/dioxide. Specific inhibitors of ATM (KU55933) or HSP90 (17AAG) were found to increase the Th- cytotoxicity suggesting prosurvival role of these signaling molecules in rescuing the cells from Th-toxicity.

Size-dependent toxicity of ThO2 nanoparticles to green algae Chlorella pyrenoidosa.

Thorium (Th) is a natural radioactive element present in the environment and has the potential to be used as a clean nuclear fuel. Relatively little is known about the aquatic toxicity of Th, especially in nanoparticulate form, which may be the main chemical species of Th in the natural waters. In this study, impacts of ThO2 nanoparticles (NPs) with two different sizes (52 ± 5 nm, s-ThO2vs. 141 ± 6 nm, b-ThO2) on a green alga Chlorella pyrenoidosa (C. pyrenoidosa) were evaluated. Results indicated that C. pyrenoidosa was more sensitive to s-ThO2 (96-h EC30 = 64.1 µM) than b-ThO2 (96-h EC30 = 100.2 µM). Exposure to 200 µM of ThO2 NPs reduced the chlorophyll-a and chlorophyll-b contents of the algal cells. At 96 h, SEM and TEM showed that more agglomerates of s-ThO2 than those of b-ThO2 were attached onto the surface of algal cells. Reactive oxygen species (ROS) generation and membrane damage were induced after the attachment of high concentrations of ThO2 NPs. The heteroagglomeration between ThO2 NPs and algal cells and increased oxidative stress might play important roles in the toxicity of ThO2 NPs. To the best of our knowledge, this is the first report on aquatic toxicity of ThO2 NPs.

Radiation databases and archives - examples and comparisons.

Studies of ionizing radiation effects through the archiving of data began with standardizing medical treatments in the early 1900s shortly after the discovery of X-rays. Once the breadth of the delayed effects of ionizing radiation was recognized, the need for long-term follow up became apparent. There are now many human archives of data from nuclear disasters and accidents, occupational exposures, and medical procedures. Planned animal irradiation experiments began around the time of the Cold War and included a variety of doses, fractions, dose rates, and types of ionizing radiation. The goal of most of these studies was to supplement information coming from human data through carefully planned experimental conditions and immediate and uninterrupted data collection. This review aims to highlight major archives and databases that have shaped the field of radiation biology and provide a broad range of the types of datasets currently available. By preserving all of these data and tissue sets, radiation biologists can combine databases and conduct large-scale analyses of detailed existing data and perform new assays with cutting edge scientific approaches.

[Retention of contrast media in the history of radiology : Sequelae of the former use of thorotrast and new challenges]. / Kontrastmittelretention in der Geschichte der Radiologie : Folgen der früheren Thorotrastanwendungen und neue Herausforderungen.

Detection of gadolinium deposits in patients who have repeatedly been administered intravenous gadolinium chelates have given rise to concern regarding the long-term safety of magnetic resonance imaging (MRI) contrast media. Nevertheless, negative long-term clinical effects have not yet been observed. In some publications parallels have been drawn to the sequelae of thorotrast that was formerly used for arterial angiography. In this article the history of thorotrast use is briefly described and in particular why, despite warnings, this substance was used frequently and worldwide. A brief summary of the results of the German Thorotrast Study revealed that high excess rates were only observed for primary malignant liver tumors after a 15-year or longer latency period and to a lesser degree of leukemias, as well as for severe local complications due to paravascular injections, particularly in the neck region. Based on this historical review, we will venture to take stock of the outcome from the "success story" of this contrast agent.

The German Thorotrast Cohort Study: a review and how to get access to the data.

It is well known that exposures like those from (226)Ra, (224)Ra and Thorotrast(®) injections increase the risk of neoplasia in bone marrow and liver. The thorium-based radioactive contrast agent Thorotrast(®) was introduced in 1929 and applied worldwide until the 1950s, especially in angiography and arteriography. Due to the extremely long half-life of several hundred years and the life-long retention of the thorium dioxide particles in the human body, patients suffer lifetime internal exposure. The health effects from the incorporated thorium were investigated in a few cohort studies with a German study being the largest among them. This retrospective cohort study was set up in 1968 with a follow-up until 2004. The study comprises 2326 Thorotrast patients and 1890 patients of a matched control group. For those being alive at the start of the study in 1968 follow-up was done by clinical examinations on a biannual basis. For the others, causes of death were collected in various ways. Additionally, clinical, radiological and biophysical studies of patients were conducted and large efforts were made to best estimate the radiation doses associated with incorporation of the Thorotrast. The aim of this paper is to describe the cohort, important results and some open questions. The data from the German Thorotrast Study are available to other interested researchers. Information can be found at http://storedb.org .

Inorganic arsenic speciation by differential pulse anodic stripping voltammetry using thoria nanoparticles-carbon paste electrodes.

Two novel thoria (ThO2) nanoparticles-carbon paste electrodes were used to evaluate an anodic stripping voltammetric method for the direct determination of arsenite and total inorganic arsenic (arsenite plus arsenate) in water samples. The effect of Ag((I)), Cu((II)), Hg((II)), Sb((III)) and Se((IV)) ions on the electrochemical response of arsenic was assayed. The developed electroanalytical method offers a rapid procedure with improved analytical characteristics including good repeatability (3.4%) at low As((III)) concentrations, high selectivity, lower detection limit (0.1 µg L(-1)) and high sensitivity (0.54 µA µg(-1) L). The analytical capability of the optimized method was demonstrated by the determination of arsenic in certified reference materials (trace elements in natural water, trace elements in water and coal fly ash).

Visualization of the glomerular endothelial glycocalyx by electron microscopy using cationic colloidal thorium dioxide.

Biological material itself appears with poor contrast in electron microscopy (EM), due to its composition mostly of light elements. Classical staining agents such as osmium tetroxide, uranyl acetate, and lead citrate preserve and/or stain cellular structures such as membranes, cytoplasm, and organelles well for EM. However, extracellular polymeric substances (EPS) show no or only poor contrast with these staining agents. The endothelial glycocalyx in blood vessels consists mainly of proteoglycans. It can be visualized by EM only by additional staining with heavy metal ions such as copper (Alcian blue, cupromeronic blue), ruthenium (ruthenium red), or lanthanum. Best results are achieved by combined perfusion of fixative and stain. Cationic hydrous thorium dioxide colloids (named here cThO2) trace acidic groups in EPS. We describe here the use of cThO2 to visualize the glomerular endothelial glycocalyx in the mouse kidney. cThO2 shows high electron density and binds to a continuous layer of up to a few hundred nanometers thickness on the glomerular endothelium, as well as on epithelia in other blood vessels in perfused animals. The observed staining pattern gives rise to periodic densities, with a spacing varying between 50 and 200 nm, depending on the overall layer thickness, which varies between below 50 up to 300 nm. Due to high electron density of the used cThO2 particles, the introduced method allows distinct imaging and precise fine structural analysis of the endothelial glycocalyx.

Radiation pathology: from thorotrast to the future beyond radioresistance.

The effects of radiation on human health have been a major concern, especially after the Fukushima Daiichi Nuclear Power Plant (FNPP) accident. We can determine these effects only from radiological disasters. The radiological contrast medium Thorotrast is known to induce hepatic cancers decades after injection. Using archival materials from Thorotrast patients, we performed molecular pathological studies to elucidate carcinogenic mechanisms of internal radiation exposure. It is emphasized here that radiation-induced cancers are a complex consequence of biological response to radiation and ingested radionuclides. We further expanded the study to establish clinically relevant radioresistant cancer cells in order to develop more effective and less harmful radiation therapy. We also found that cancer cells can acquire radioresistance by low-dose fractionated radiation within one month. The FNPP accidents prompted us to collect tissue samples from animals in and around the evacuation zone in order to construct a tissue bank. The final goal of the bank is to enable research that will contribute to the common understanding of radioprotection.

Thorotrast and in vivo thorium dioxide: numerical simulation of 30 years of α radiation absorption by the tissues near a large compact source.

BACKGROUND: The epidemiology of the slightly radioactive contrast agent named Thorotrast presents a very long latency period between the injection and the development of the related pathologies. It is an example of the more general problem posed by a radioactive internal contaminant whose effects are not noteworthy in the short term but become dramatic in the long period. A point that is still to be explored is fluctuations (in space and time) in the localized absorption of radiation by the tissues. METHODS: A Monte Carlo simulation code has been developed to study over a 30-year period the daily absorption of α radiation by µm-sized portions of tissue placed at a distance of 0-100 µm from a model source, that approximates a compact thorium dioxide source in liver or spleen whose size is ≳20 µm. The biological depletion of the daughter nuclei of the thorium series is taken into account. The initial condition assumes chemically purified natural thorium. RESULTS: Most of the absorbed dose is concentrated in a 25-µm thick layer of tissue, adjacent to the source boundary. Fluctuations where a target region with a volume of 1 µm(3) is hit by 3-5 α particles in a day or in a shorter period of time are relevant in a 1-10 µm thick layer of tissue adjacent to the source boundary, where their frequency is larger than the Poisson-law prediction.

Hepatic angiosarcoma occurring 65 years after thorium dioxide (Thorotrast) exposure: imaging, surgical and histo- pathologic findings of a historical case.

We report the CT, surgical and histopathologic findings of a rare case of Hepatic Angiosarcoma (HAS) diagnosed in a 85-year old women 65 years afterThorotrast (Th232) exposure for angiography. At the early arterial phase of dynamic MDCT, peripheral curvilinear and central nodular puddling of contrast produced in the 8 cm tumor.Then progressive contrast filling of the tumor was observed on the delayed scans. Associated pathognomonic signs related to previous Th232 exposure were also found comprising diffuse intrahepatic reticular bands of calcifications, numerous calcified epigastric lymph nodes and a calcified shrunken spleen. Emergency laparotomy was performed because of associated hemoperitoneum. With a delay of 65 years afterThorotrast exposure, this historical case probably represents, to our knowledge, the most delayed presentation of Th232 related HAS ever published.

Incidental hyperdensities within the reticuloendothelial system.

This is a case report of a 79-year-old woman who was found to have numerous hyperdense nodular lesions in the upper abdomen, which were incidentally discovered during routine follow-up of a lung nodule. These hyperdense lesions included a lace-like reticular distribution within the liver, multiple extremely dense lymph nodes, and a shrunken hyperdense spleen. We discuss differential considerations for such a constellation of findings and explain why we believe the findings in this case are consistent with prior thorium dioxide exposure. We conclude with a discussion of the pathophysiology and important complications of thorium dioxide exposure and the best imaging modalities for its detection. We believe that this is an important entity that all physicians should be aware of because even though it is seldom seen today, it has characteristic imaging findings and the correct diagnosis is critical given the increased risk of malignancy for which such patients should be screened for.

Thorotrast: analysis of the time evolution of its α activity concentration, in the 70 years following the chemical purification of Thorium.

We simulate the α-activity of the Thorium series elements present in the contrast medium named Thorotrast, used until 1960 and cause of certified deaths until today. Assuming, as active components at t=0, (232)Th and (228)Th in the same relative concentration they have in nature, α-activity oscillates for some decades before reaching a stationary value that in absence of biological depletion would be AST =24000Bq/g. Our Montecarlo code generates the nuclear decays of the Thorium series with and without in-vivo biological depletion, arriving to three kinds of results for the activity: 1) Theoretical activity concentration (no biological depletion). Our result is fitted by: A(t)=A(ST).{[1-exp(-t/10)]+[exp(-t/tB)(1-0.8exp(-t/tA))]}, with t in years, tA=1.07.10(-2) years, and tB=2.38 years. 2) Weak biological depletion (228Ra/232 Th equilibrium activity ratio 0.6, 224Ra/228Ra e.a.r 0.9, 10% excretion for 220Rn). The ratio of the activity concentration to the theoretical activity concentration is fitted by: A weak (t)/A(t)=0.61+0.29 exp[-(t/15)2] (t in years). 3) Strong biological depletion (228Ra/232Th e.a.r 0.4, 224Ra/228Ra e.a.r. 0.8, 10% excretion for 220Rn). The ratio of the activity concentration to the theoretical activity concentration is fitted by A(strong)(t)/A(t)=0.44+0.4 exp[-(t/13)2](t in years). We also report fluctuation calculation for two cases where standard statistical behavior is not expected.