High prevalence of mutant KRAS in circulating exosome-derived DNA from early-stage pancreatic cancer patients.

Background: Exosomes arise from viable cancer cells and may reflect a different biology than circulating cell-free DNA (cfDNA) shed from dying tissues. We compare exosome-derived DNA (exoDNA) to cfDNA in liquid biopsies of patients with pancreatic ductal adenocarcinoma (PDAC). Patients and methods: Patient samples were obtained between 2003 and 2010, with clinically annotated follow up to 2015. Droplet digital PCR was performed on exoDNA and cfDNA for sensitive detection of KRAS mutants at codons 12/13. A cumulative series of 263 individuals were studied, including a discovery cohort of 142 individuals: 68 PDAC patients of all stages; 20 PDAC patients initially staged with localized disease, with blood drawn after resection for curative intent; and 54 age-matched healthy controls. A validation cohort of 121 individuals (39 cancer patients and 82 healthy controls) was studied to validate KRAS detection rates in early-stage PDAC patients. Primary outcome was circulating KRAS status as detected by droplet digital PCR. Secondary outcomes were disease-free and overall survival. Results: KRAS mutations in exoDNA, were identified in 7.4%, 66.7%, 80%, and 85% of age-matched controls, localized, locally advanced, and metastatic PDAC patients, respectively. Comparatively, mutant KRAS cfDNA was detected in 14.8%, 45.5%, 30.8%, and 57.9% of these individuals. Higher exoKRAS MAFs were associated with decreased disease-free survival in patients with localized disease. In the validation cohort, mutant KRAS exoDNA was detected in 43.6% of early-stage PDAC patients and 20% of healthy controls. Conclusions: Exosomes are a distinct source of tumor DNA that may be complementary to other liquid biopsy DNA sources. A higher percentage of patients with localized PDAC exhibited detectable KRAS mutations in exoDNA than previously reported for cfDNA. A substantial minority of healthy samples demonstrated mutant KRAS in circulation, dictating careful consideration and application of liquid biopsy findings, which may limit its utility as a broad cancer-screening method.

Interpreting biomarker data from the COPHES/DEMOCOPHES twin projects: Using external exposure data to understand biomarker differences among countries.

In 2011 and 2012, the COPHES/DEMOCOPHES twin projects performed the first ever harmonized human biomonitoring survey in 17 European countries. In more than 1800 mother-child pairs, individual lifestyle data were collected and cadmium, cotinine and certain phthalate metabolites were measured in urine. Total mercury was determined in hair samples. While the main goal of the COPHES/DEMOCOPHES twin projects was to develop and test harmonized protocols and procedures, the goal of the current paper is to investigate whether the observed differences in biomarker values among the countries implementing DEMOCOPHES can be interpreted using information from external databases on environmental quality and lifestyle. In general, 13 countries having implemented DEMOCOPHES provided high-quality data from external sources that were relevant for interpretation purposes. However, some data were not available for reporting or were not in line with predefined specifications. Therefore, only part of the external information could be included in the statistical analyses. Nonetheless, there was a highly significant correlation between national levels of fish consumption and mercury in hair, the strength of antismoking legislation was significantly related to urinary cotinine levels, and we were able to show indications that also urinary cadmium levels were associated with environmental quality and food quality. These results again show the potential of biomonitoring data to provide added value for (the evaluation of) evidence-informed policy making.

Diabetes, antidiabetic medications, and pancreatic cancer risk: an analysis from the International Pancreatic Cancer Case-Control Consortium.

BACKGROUND: Type 2 diabetes mellitus has been associated with an excess risk of pancreatic cancer, but the magnitude of the risk and the time-risk relationship are unclear, and there is limited information on the role of antidiabetic medications. PATIENTS AND METHODS: We analyzed individual-level data from 15 case-control studies within the Pancreatic Cancer Case-Control Consortium, including 8305 cases and 13 987 controls. Pooled odds ratios (ORs) were estimated from multiple logistic regression models, adjusted for relevant covariates. RESULTS: Overall, 1155 (15%) cases and 1087 (8%) controls reported a diagnosis of diabetes 2 or more years before cancer diagnosis (or interview, for controls), corresponding to an OR of 1.90 (95% confidence interval, CI, 1.72-2.09). Consistent risk estimates were observed across strata of selected covariates, including body mass index and tobacco smoking. Pancreatic cancer risk decreased with duration of diabetes, but a significant excess risk was still evident 20 or more years after diabetes diagnosis (OR 1.30, 95% CI 1.03-1.63). Among diabetics, long duration of oral antidiabetic use was associated with a decreased pancreatic cancer risk (OR 0.31, 95% CI 0.14-0.69, for ≥15 years). Conversely, insulin use was associated with a pancreatic cancer risk in the short term (OR 5.60, 95% CI 3.75-8.35, for <5 years), but not for longer duration of use (OR 0.95, 95% CI 0.53-1.70, for ≥15 years). CONCLUSION: This study provides the most definitive quantification to date of an excess risk of pancreatic cancer among diabetics. It also shows that a 30% excess risk persists for more than two decades after diabetes diagnosis, thus supporting a causal role of diabetes in pancreatic cancer. Oral antidiabetics may decrease the risk of pancreatic cancer, whereas insulin showed an inconsistent duration-risk relationship.

Indoor air pollution from solid fuels and risk of upper aerodigestive tract cancers in central and eastern Europe.

BACKGROUND: Indoor air pollution from solid fuels is a potentially important risk factor for cancer, yet data on cancers from organs other than the lung are scarce. We investigated if indoor air pollution from coal and wood are risk factors for additional cancers, particularly that of the upper aerodigestive tract (oral cavity, larynx, pharynx and esophagus) in the high-risk areas of central and eastern Europe. METHODS: We used data from multi-center hospital-based case-control study of 1065 histologically confirmed upper aerodigestive tract cancer cases and 1346 controls. Standardized questionnaires were used to collect information on residential fuel use for cooking and heating. Using unconditional logistic regression, we calculated odds ratios (ORs) and 95% confidence intervals (95% CIs) for upper aerodigestive tract cancer risk after adjusting for potential confounders. RESULTS: Lifelong wood use was associated with pharyngeal and esophageal (OR 4.05, 95% CI: 1.30-12.68 and OR 2.71, 95% CI: 1.21-6.10, respectively). We observed an exposure-response relationship between duration of wood use and risk of pharyngeal cancer among those who had never used coal (P(trend)=0.04), ruling out the possibility of residual confounding by coal. Similarly, we observed an increased risk of laryngeal cancers and head & neck cancers among those who always used coal, with a noted exposure-response relationship (P(trend)<0.01). CONCLUSIONS: Our results suggest a possible role of indoor air pollution from solid fuel use in head and neck carcinogenesis in the high risk area of central and eastern Europe.

Dietary risk factors for squamous cell carcinoma of the upper aerodigestive tract in central and eastern Europe.

OBJECTIVE: The incidence of squamous cell carcinoma of upper aerodigestive tract (UADT: oral cavity, pharynx, larynx, and esophagus) has been increasing in central and eastern European countries. We investigated the relationship between diet and UADT cancers in these high risk areas. METHODS: We used data from hospital-based case-control study of 948 UADT cancer cases and 1,228 controls conducted in Romania, Hungary, Poland, Russia, Slovakia, and Czech Republic. Standardized questionnaire were used to collect information on 23 different food items, along with alcohol and tobacco consumptions. Logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for the UADT cancers after adjusting for center, age, sex, tobacco & alcohol intake, and other food groups. RESULTS: Consumption of dairy product was negatively associated with selected UADT cancers: larynx (OR: 0.38, CI: 0.23-0.62) and esophagus (OR: 0.55, CI: 0.33-0.93). While consumption of yellow/orange vegetables were inversely associated with oral/pharyngeal and laryngeal cancer (OR: 0.53, CI: 0.35-0.81 and OR: 0.62, CI: 0.38-1.00, respectively), preserved vegetable was positively associated with oral/pharyngeal and laryngeal cancer risk (p (trend) < 0.01 for both). CONCLUSION: Specific dietary components may play a role in the development of UADT cancers in the high-risk region of central and eastern Europe.

Chromosomal aberrations and SCEs as biomarkers of cancer risk.

Previous studies have suggested that the frequency of chromosomal aberrations (CAs), but not of sister chromatid exchanges (SCEs), predicts cancer risk. We have further examined this relationship in European cohorts comprising altogether almost 22,000 subjects, in the framework of a European collaborative project (CancerRiskBiomarkers). The present paper gives an overview of some of the results of the project, especially as regards CAs and SCEs. The results confirm that a high level of CAs is associated with an increased risk of cancer and indicate that this association does not depend on the time between CA analysis and cancer detection, i.e., is obviously not explained by undetected cancer. The present evidence indicates that both chromatid-type and chromosome-type CAs predict cancer, even though some data suggest that chromosome-type CAs may have a more pronounced predictive value than chromatid-type CAs. CA frequency appears to predict cancers at various sites, although there seems to be a particular association with gastrointestinal cancers. SCE frequency does not appear to have cancer predictive value, at least partly due to uncontrollable technical variation. A number of genetic polymorphisms of xenobiotic metabolism, DNA repair, and folate metabolism affect the level of CAs and might collectively contribute to the cancer predictivity of CAs. Other factors that may influence the association between CAs and cancer include, e.g., exposure to genotoxic carcinogens and internal generation of genotoxic species. Although the association between CA level and cancer is seen at the group level, an association probably also exists for the individual, although it is not known if an individual approach could be feasible. However, group level evidence should be enough to support the use of CA analysis as a tool in screening programs and prevention policies in occupational and environmental health.

Parental education and children's respiratory and allergic symptoms in the Pollution and the Young (PATY) study.

Inequalities in health between socio-economic groups are a major public health concern. The current authors studied associations between parental socio-economic status (SES) and children's respiratory and allergic symptoms in 13 diverse countries, including the Russian Federation, North America (Canada and the USA), and countries across Eastern and Western Europe. Data of 57,000 children aged 6-12 yrs, originating from eight cross-sectional studies, were analysed. SES was defined by parental education. Respiratory and allergic symptoms were defined by parental questionnaire reports. Multiple logistic regressions showed that low parental education was associated with a decreased risk of inhalant allergy and itchy rash in school children. Furthermore, low parental education was associated with an increased prevalence of wheeze and nocturnal dry cough. No clear association was found between parental education and prevalence of doctor-diagnosed asthma and bronchitis. Part of the difference between socio-economic groups with regard to their children's symptoms was explained by established risk factors, such as parental allergy, smoking during pregnancy, pet ownership, crowding, mould/moisture in the home, use of gas for cooking, and air pollution (particulate matter with a diameter of <10 microm). However, differences remained after adjusting for these variables. Children's health was associated with parental education. The association could not fully be explained by established risk factors.

Respiratory symptoms, bronchitis and asthma in children of Central and Eastern Europe.

The multicentre Central European Study of Air Pollution and Respiratory Health (CESAR) aimed to measure the respiratory health of schoolchildren using a standardised questionnaire in six countries of Central and Eastern Europe (CEE), allowing comparisons within this region and with other European countries. A cross-sectional study was conducted in 25 urban areas of Bulgaria, Czech Republic, Hungary, Poland, Romania, and Slovakia in 1996. Parents of 21,743 schoolchildren of age 7-11 yrs completed a questionnaire based on items from the World Health Organization and International Study of Asthma and Allergies in Childhood questions on cough and wheeze symptoms, as well as on diagnoses by doctors. Life-time prevalence of bronchitis was 55.9%, asthma 3.9%, and asthmatic, spastic or obstructive bronchitis 12.3%. In CEE countries the prevalence of bronchitis is higher and prevalence of asthma appears lower than in Western Europe. However, if asthma is defined as a diagnosis of either asthma or asthmatic, spastic or obstructive bronchitis, then its prevalence is comparable to Western Europe, or higher. In this region, within-country variation for most respiratory parameters is less than between-country variation. Between-country comparisons in doctors' diagnoses appear dependent on the choice of definition of asthma. Europe-wide comparisons in prevalence of respiratory symptoms and diagnosis are reported in this study. Some of the East-West difference in asthma prevalence may be attributable to differences in diagnostic practice.

Childhood respiratory symptoms, hospital admissions, and long-term exposure to airborne particulate matter.

The effects of long-term exposure to air pollution on respiratory symptoms and respiratory hospitalization (for asthma, bronchitis or pneumonia) were assessed in a cross-sectional study of children (ages 7--11 years, N=667) living in a moderately industrialized city in Central Slovakia. Individual health, residence and family history data obtained through the CESAR study were coupled, using Geographic Information System (GIS) technologies, with total suspended particulate (TSP) exposure estimates derived from dispersion modeling of almost all local stationary sources. These data were used to assess, at the intra-city level and child-specific level, the potential for TSP as a risk factor for respiratory disease in children. TSP, PM10, and PM2.5 monitored ambient concentrations are highly correlated in the study location. Modeled TSP concentrations resulting from local source emissions are dominated by a large wood processing facility, suggesting variation in exposures among children. The prevalence of respiratory non-asthmatic symptoms and hospitalizations was associated with increased TSP. No association between long-term exposure to TSP and asthma diagnosis or wheeze symptoms was found. Logistic regression modeling indicated a significant increase in hospital admissions for asthma, bronchitis or pneumonia associated with increasing air pollution (OR 2.16, CI, 1.01--4.60), doctor-diagnosed bronchitis (OR 1.53, CI, 1.02-2.30), and parent-reported chronic phlegm (OR 3.43, CI, 1.64--7.16), expressed as odds for a 15 microg/m3 increase in estimated TSP exposure, and these increases are not due to differences in socio-economic, health care or other identified factors.

Personal exposure to particles in Banská Bystrica, Slovakia.

Epidemiological studies have associated adverse health impacts with ambient concentrations of particulate matter (PM), though these studies have been limited in their characterization of personal exposure to PM. An exposure study of healthy nonsmoking adults and children was conducted in Banska Bystrica, Slovakia, to characterize the range of personal exposures to air pollutants and to determine the influence of occupation, season, residence location, and outdoor and indoor concentrations on personal exposures. Twenty-four-hour personal, at-home indoor, and ambient measurements of PM10, PM2.5, sulfate (SO4(2-)) and nicotine were obtained for 18 office workers, 16 industrial workers, and 15 high school students in winter and summer. Results showed that outdoor levels of pollutants were modest, with clear seasonal differences: outdoor PM10 summer/winter mean = 35/45 microg/m3; PM2.5 summer/winter mean = 22/32 microg/m3. SO4(2-) levels were low (4-7 microg/m3) and relatively uniform across the different sample types (personal, indoor, outdoor), areas, and occupational groups. This suggests that SO4(2-) may be a useful marker for combustion mode particles of ambient origin, although the relationship between personal exposures and ambient SO4(2-) levels was more complex than observed in North American settings. During winter especially, the central city area showed higher concentrations than the suburban location for outdoor, personal, and indoor measures of PM10, PM2.5, and to a lesser extent for SO4(2-), suggesting the importance of local sources. For PM2.5 and PM10, ratios consistent with expectations were found among exposure indices for all three subject groups (personal>indoor>outdoor), and between work type (industrial>students>office workers). The ratio of PM2.5 personal to indoor exposures ranged from 1.0 to 3.9 and of personal to outdoor exposures from 1.6 to 4.2. The ratio of PM10 personal to indoor exposures ranged from 1.1 to 2.9 and the ratio of personal to outdoor exposures from 2.1 to 4.1. For a combined group of office workers and students, personal PM10/PM2.5 levels were predicted by statistically significant multivariate models incorporating indoor (for PM2.5) or outdoor (for PM10) PM levels, and nicotine exposure (for PM10). Small but significant fractions of the overall variability, 15% for PM2.5 and 17% for PM10, were explained by these models. The results indicate that central site monitors underpredict actual human exposures to PM2.5 and PM10. Personal exposure to SO4(2-) was found to be predicted by outdoor or indoor SO4(2-) levels with 23-71% of the overall variability explained by these predictors. We conclude that personal exposure measurements and additional demographic and daily activity data are crucial for accurate evaluation of exposure to particles in this setting.

Health risk assessment for inhalation exposure to arsenic.

Health risk assessment was used as the formal process to estimate the likelihood and magnitude of the health effects occurring in humans as a result of environmental and occupational exposure to polluting agents. This study was focused at estimating the human health risk of the general and working population living in the region polluted by arsenic for more than 40 years, from combustion of coal with high arsenic content in the power plant. The exposure to arsenic from inhalation was under investigation. A study period of 40 years (1973-1993) was chosen. The study period was defined taking into account, besides the availability of data, the temporal patterns of the technological processes and the trends over time of environmental concentrations. The results from the arsenic risk assessment study were used for the evaluation of the health risk for different population groups in the polluted areas and for different professions of workers exposed to As in a power plant. The results are applicable for the evaluation of risk in real conditions, for health surveillance and for remedial changes and a potential suggestion on technological improvement.

Immune biomarkers in relation to exposure to particulate matter: a cross-sectional survey in 17 cities of Central Europe.

Human population data on air pollution and its effects on the immune system are scarce. A survey was conducted within the framework of the Central European Study of Air Quality and Respiratory Health (CESAR) to measure a panel of immune biomarkers in children of Bulgaria, Czech Republic, Hungary, Poland, Romania, and Slovakia. Seventeen cities were chosen to represent a wide range of exposure to outdoor air pollution. In each, ambient particulate matter of less than 10 microns diameter and less than 2.5 microns diameter (PM10 and PM2.5) were measured with a Harvard impactor. Blood was collected from 366 school children aged 9 to 11 yr between 11 April and 10 May 1996. The percentage of B, total T, CD4+, CD8+, and natural killer (NK) lymphocytes was determined by flow cytometry (Becton Dickinson); total immunoglobulins of class G, M, A and E (IgG, IgM, IgA, and IgE) were measured in serum using nephelometry (Behring). Associations between PM and each log-transformed biomarker concentration were studied by linear regression, in a two-stage model. The yearly average concentrations varied from 41 to 96 micrograms/m3 for PM10 across the 17 study areas, from 29 to 67 micrograms/m3 for PM2.5, and from 12 to 38 micrograms/m3 for PM10-2.5 (coarse). Number of B, CD4+, CD8+, and NK lymphocytes increased with increasing concentration of PM, having adjusted for age, gender, parental smoking, laboratory of analysis, and recent respiratory illness. Differences in lymphocyte number were larger and statistically significant for exposure to PM2.5. Similar results were found when we examined the association between PM and lymphocyte number separately for each laboratory. Total IgG was increased with increasing concentration of PM, significantly in the case of PM2.5. When we repeated the analyses with two other statistical approaches the results did not differ from those reported here. The effect of coarse PM on lymphocyte numbers appears small in comparison to PM2.5. One possible interpretation of our findings is that long-term exposure to airborne particulates leads to inflammation of the airways and activation of the cellular and humoral immune system.

Occupational cancer in central European countries.

The countries of central Europe, including Poland, the Czech Republic, Slovakia, Hungary, Romania, and Bulgaria, suffer from environmental and occupational health problems created during the political system in place until the late 1980s. This situation is reflected by data on workplace exposure to hazardous agents. Such data have been systematically collected in Skovakia and the Czech Republic since 1977. The data presented describe mainly the situation in the early 1990s. The number of workers exposed to risk factors at the workplace represent about 10% of the working population in Slovakia and 30% in Poland. In Slovakia in 1992 the percentage of persons exposed to chemical substances was 16.4%, to ionizing radiation 4.3%, and to carcinogens 3.3% of all workers exposed to risk factors. The total number of persons exposed to substances proven to be carcinogens in Poland was 1.3% of the employees; 2.2% were exposed to the suspected carcinogens. The incidence of all certified occupational diseases in the Slovak Republic was 53 per 100,000 insured employees in 1992. Cancers certified as occupational cancers are skin cancer caused by occupational exposure to carcinogens, lung cancer caused by ionizing radiation, and asbestosis together with lung cancer. Specific information on occupational cancers from Romania and Bulgaria was not available for this paper. It is difficult to predict a trend for future incidences of occupational cancer. Improved control technology, governmental regulatory activity to reduce exposure, surveillance of diseases and risk factors, and vigilant use of preventive measures should, however, ultimately reduce occupational cancer.

Health status of workers of a thermal power station exposed for prolonged periods to arsenic and other elements from fuel.

The Nováky Power Station (NPS) has been using since 1953 as fuel coal with a high content of As and with a low content of other metals. This involves a constant risk for the workers as well as pollution of the surroundings. The authors described 16 cases of chronic As intoxication in NPS workers after 22.3 +/- 8.4 years of exposure (especially stokers, maintenance workers, boiler cleaners). Among clinical symptoms prevailed sensory and motor polyneuropathy (13 cases), pseudoneurasthenic syndrome (10 cases), toxic encephalopathy (6 cases) and nasal septum perforation (2 cases). After 1989 the intoxications with As did not occur any more due to technical measures and health protection of the workers. The authors present a review of actual results of clinical, haematological and biochemical investigations and tests for metals (AAS methods) in biological materials of workers at risk in NPS (n = 70), exposed on average for 11.9 +/- 0.5 years, of average age 35.91 +/- 1.7 years (mean +/- SE) and compared the results to a matched control group of blood donors not exposed to metals (n = 29). In NPS workers significantly lower Hb values, higher serum creatinine, higher serum beta 2-microglobulin, higher As content in hair as well as higher serum Mn and Pb concentrations compared with the C-group were found. The exposed group had significantly lower serum Se concentrations (0.64 +/- 0.025 mumol/l (mean +/- SE) compared to Se levels of persons from an adjacent district. The authors stress the necessity of individual evaluation of the metal concentrations in relation to clinical findings. With prolonged exposure the situation can become more urgent not only because of chronic poisoning but also because of the cancerogenic effects of these elements on the human organism.

Airborne arsenic and urinary excretion of arsenic metabolites during boiler cleaning operations in a Slovak coal-fired power plant.

Little information is available on the relationship between occupational exposure to inorganic arsenic in coal fly ash and urinary excretion of arsenic metabolites. This study ws undertaken in a coal-fired power plant in Slovakia during a routine maintenance outage. Arsenic was measured in the breathing zone of workers during 5 consecutive workdays, and urine samples were obtained for analysis of arsenic metabolites--inorganic arsenic (Asi), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA)--prior to the start of each shift. Results from a small number of cascade impactor air samples indicated that approximately 90% of total particle mass and arsenic was present in particle size fractions >/= 3.5 micron. The 8-hr time-weighted average (TWA) mean arsenic air concentration was 48.3 microg/m3 (range 0.17-375.2) and the mean sum of urinary arsenic (SigmaAs) metabolites was 16.9 microg As/g creatinine (range 2.6-50.8). For an 8-hr TWA of 10 microg/m3 arsenic from coal fly ash, the predicted mean concentration of the SigmaAs urinary metabolites was 13.2 microg As/G creatinine [95% confidence interval (CI), 10.1-16.3). Comparisons with previously published studies of exposure to arsenic trioxide vapors and dusts in copper smelters suggest that bioavailability of arsenic from airborne coal fly ash (as indicated by urinary excretion) is about one-third that seen in smelters and similar settings. Arsenic compound characteristics, matrix composition, and particle size distribution probably play major roles in determining actual uptake of airborne arsenic.

Cytogenetic analysis of peripheral blood lymphocytes in workers exposed to vinyl chloride.

In the present study, the method of cytogenetic analysis of peripheral blood lymphocytes was used to examine 43 workers exposed to vinyl chloride monomer (average exposure 11.2 years) and 22 subjects selected from the same locality (control group). A total number of 8650 metaphases were analysed. All cytogenetic parameters examined were increased in the exposed group as compared to the control group and 3 parameters, chromatid breaks, percentage of aberrant cells and breaks per cell, were significantly increased (P less than 0.001).