Raffaella Uccelli

Predictions of mortality from pleural mesothelioma in Italy: A model based on asbestos consumption figures supports results from age-period-cohort models

Italy was the second main asbestos producer in Europe, after the Soviet Union, until the end of the 1980s, and raw asbestos was imported on a large scale until 1992. The Italian pattern of asbestos consumption lags on average about 10 years behind the United States, Australia, the United Kingdom and the Nordic countries. Measures to reduce exposure were introduced in the mid‐1970s in some workplaces. In 1986, limitations were imposed on the use of crocidolite and in 1992 asbestos was definitively banned. We have used primary pleural cancer mortality figures (1970–1999) to predict mortality from mesothelioma among Italian men in the next 30 years by age‐cohort‐period models and by a model based on asbestos consumption figures. The pleural cancer/mesothelioma ratio and mesothelioma misdiagnosis in the past were taken into account in the analysis. Estimated risks of birth cohorts born after 1945 decrease less quickly in Italy than in other Western countries. The findings predict a peak with about 800 mesothelioma annual deaths in the period 2012–2024. Results estimated using age‐period‐cohort models were similar to those obtained from the asbestos consumption model.

An exploratory case-control study on spinal and bulbar forms of amyotrophic lateral sclerosis in the province of Rome

Several environmental and life-style factors reported as possibly associated with ALS have been analysed in the present study, focusing on the two clinical onsets of ALS. A case-control study (77 cases and 185 controls) has been performed in the province of Rome in the period 2005–2006. Increased risks were observed in bulbar cases for former smokers (OR: 4.55, 90% CI 1.72–12.08) and more than 24 pack-years, compared with spinal cases for employment in the construction sector and professional exposure to building materials (OR: 5.27, 90% CI 1.15–24.12) and metals (OR: 2.94, 90% CI 1.20–7.21). Overall and bulbar cases showed an increased risk for consumption of cold cuts and a decreased risk for vegetables intake. Regarding head injuries, differences were observed if the last injury occurred in the age range of 30–40 years, among all (OR: 14.2, 90% CI 1.04–194.42) and bulbar (OR: 17.4, 90% CI 1.70–178.5) cases, and less than 30 years among spinal cases (OR: 7.13, 90% CI 1.34–37.94). Moreover, a risk for a time period of 11–30 years since the last head injury suffered was found in bulbar cases (OR: 3.51, 90% CI 1.03–11.95). Some of the hypothesized risk factors for ALS have been found positively associated in this study, with different patterns between bulbar and spinal ALS.

Flow Cytometric Analysis of the Effects of 0·4 MeV Fission Neutrons on Mouse Spermatogenesis

(C57Bl/Cne X C3H/Cne)F1 male mice were irradiated with single acute doses of 0.4 MeV neutrons ranging from 0.05 to 2 Gy, and testis cell suspensions were prepared for cytometric analysis of the DNA content 2-70 days after irradiation. Various cell subpopulations could be identified in the control histogram including mature and immature spermatids, diploid spermatogonia and spermatocytes, tetraploid cells and cells in the S-phase. Variations in the relative proportions of different cell types were detected at each dose and time, reflecting lethal damage induced on specific spermatogenetic stages. The reduction of the number of elongated spermatids 28 days after irradiation was shown to be a particularly sensitive parameter for the cytometrical assessment of the radiosensitivity of differentiating gonia. A D0 value of 0.13 Gy was calculated and compared with data obtained after X-irradiation, using the same experimental protocol. In the latter case a biphasic curve was obtained over the dose range from 0.25 to 10 Gy, possibly reflecting the existence of some cell population heterogeneity. RBE values were estimated at different neutron doses relative to the radiosensitive component of the X-ray curve, and ranged from 3.3 to 4, in agreement with data in the literature. Genotoxic effects were monitored 7 days after irradiation by a dose-dependent increase of the coefficient of variation (CV) values of the round spermatid peak, reflecting the induction of numerical and structural chromosome aberrations, and 14 or 21 days after irradiation by the detection of diploid elongated spermatids, probably arising from a radiation-induced complete failure of the first or second meiotic division.

Cytotoxic effects of benzene on mouse germ cells determined by flow cytometry

Flow cytometric (FCM) DNA content measurements were performed on testicular monocellular suspensions obtained from mice exposed per os to 0, 1, 2, 4, 6, and 7 ml/kg body weight of benzene in order to investigate its cytotoxic action on germ cells. The effects of benzene were measured 7, 14, 21, 28, and 70 d after treatment. Benzene had no effect on testis weight, but FCM analysis showed the relative percentages of some cell subpopulations (tetraploid and haploid cells) to be different from the control pattern, indicating the occurrence of some cytotoxic damage to differentiating spermatogonia. These data demonstrate that spermatogenesis is sensitive to benzene single exposures as evidenced by an altered cell ratio of testicular cell types.

Effects of heat on mouse spermatogenesis monitored by flow cytometry

The effects of heat on mouse spermatogenesis have been determined using both testis weight and flow cytometrically determined DNA content distribution as experimental end-points. Temperatures of 38-42 degrees C and exposure times of 20-60 min have been tested. The results concerning the testis weight substantially confirm those reported by other authors (Hand et al. 1979). The measurement of DNA content distributions shows a relatively higher depletion, 14 days after treatment, of the cytometric compartment containing elongated spermatids in respect to that containing round spermatids. The analysis of the cytotoxic effects, monitored 14 vs. 28 days after treatment, as a function of the exposure time at a given temperature, or of the temperature for a fixed exposure time, indicates that, in the course of spermatogenesis, late spermatocytes are more sensitive to heat than differentiated spermatogonia. Following the approach based on flow cytometry, the effect of exposures as low as 20 min at 38 degrees C can be appreciated.

Retrospective mortality cohort study of Italian workers compensated for silicosis.

Objectives: To estimate cause specific mortality in a large cohort of Italian workers compensated for silicosis. Methods: The cohort included 14 929 subjects (14 098 men and 831 women) compensated for silicosis between 1946 and 1979, alive on 1 January 1980, and resident in Tuscany (a region of central Italy with 3 547 000 inhabitants). Mortality follow up ranged from 1980 to 1999. Vital status and the causes of death were determined by linkage with the regional mortality registry and with the national mortality database. The cohort mortality rates were compared to the rates of the local reference population. SMRs and their 95% confidence intervals were computed assuming a Poisson distribution of the observed deaths. Specific SMR analyses were performed according to the level of disability, the year of compensation assignment, and the job type. Results: A significant excess mortality was observed in male silicotics for cancer of the lung, trachea, and bronchus and cancer of the liver, respiratory diseases (silicosis, asbestosis, antracosilicosis, and other pneumoconiosis), and for tubercolosis. Statistically significant mortality excess was observed in female silicotics for respiratory diseases (specifically silicosis and other pneumoconiosis) and tuberculosis. Analyses for period of compensation assignment showed a twofold increased SMR for biliary tract cancer among female workers and for liver cancer among male workers compensated before 1970. Conclusions: The excess mortality from respiratory tract cancers and respiratory tract diseases detected in Italian compensated silicotics are in agreement with previous epidemiological studies. Although the twofold increased risk for liver cancer among males is suggestive of a possible association with silica dust exposure, the finding needs to be confirmed.

Exposure to Perfluoroalkyl Substances and Sperm DNA Global Methylation in Arctic and European Populations

Perfluoroalkyl substances (PFASs) are widely used in a variety of industrial processes and products, and have been detected globally in humans and wildlife. PFASs are suspected to interfere with endocrine signaling and to adversely affect human reproductive health. The aim of the present study was to investigate the associations between exposure to PFASs and sperm global methylation levels in a population of non‐occupationally exposed fertile men. Measurements of PFASs in serum from 262 partners of pregnant women from Greenland, Poland and Ukraine, were also carried out by liquid chromatography tandem mass spectrometry. Perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonic acid (PFHxS), and perfluorononanoic acid (PFNA) were detected in 97% of the blood samples. Two surrogate markers were used to assess DNA global methylation levels in semen samples from the same men: (a) average DNA methylation level in repetitive DNA sequences (Alu, LINE‐1, Satα) quantified by PCR‐pyrosequencing after bisulfite conversion; (b) flow cytometric immunodetection of 5‐methyl‐cytosines. After multivariate linear regression analysis, no major consistent associations between PFASs exposure and sperm DNA global methylation endpoints could be detected. However, since weak but statistically significant associations of different PFASs with DNA hypo‐ and hyper‐methylation were found in some of the studied populations, effects of PFASs on sperm epigenetic processes cannot be completely excluded, and this issue warrants further investigation. Environ. Mol. Mutagen. 55:591–600, 2014.

Potential Prognostic Significance of Cytometrically Determined DNA Abnormality in GI Tract Human Tumorsa

It is well known that human solid tumors are often characterized by an abnormal cellular DNA content. Recent studies after the advent of flow cytometry have confirmed that relative cellular DNA content can be a conclusive marker of malignancy. A recent review by Barlogie et al.’ has reported that cytometric aneuploidy characterizes 75% human solid tumors. In our laboratory, where flow cytometry is routinely performed for tumors of certain sites and some “precancerous” conditions, we have obtained the comparable value of 79% for over 300 cases. Flow cytometric analysis of DNA content can also be used to study some aspects of tumor cell heterogeneity in terms of ploidy level(s), occurrence of multiclonality (that is, of more than one tumor stem-cell line), and proliferation Furthermore, cytometric aneuploidy can be used in the early detection of neoplastic degeneration, especially in the instance of the GI In the present investigation, DNA flow cytometric measurements were performed on samples from 64 patients affected by GI tract tumors in the attempt to evaluate the possible prognostic significance of cytometric parameters. In fact, the possibility of exploiting flow cytometric analysis to predict the natural history of the disease andlor eventually the response to treatment has already been suggested by some authors.*-’*

Ecological conditions in the Orbetello Lagoon and suggested actions for its restoration

ABSTRACT In the Orbetello lagoon (Grosseto, Italy), the environmental impact of different anthropic activities (urban, agricultural, industrial etc.) has produced an increase in nutrient loading. It has caused instability of the ecosystem and consequently, in the last few years, recurring anoxic crises which have produced environmental damage, loss of fish and, indirectly, a decrease in tourism in the area Environmental multidisciplinary research has been carried out by ENEA with the aim of optimizing the restoration project of the lagoon and managing the transitory phase until the completion of this project. The studies have been aimed at understanding and quantifying the processes controlling the ecosystem life. To reach these goals, meteoclimatic conditions, hydrology, geochemical cycle of nutrients, biogeochemical processes, distribution and abundance of zoobenthos and macrophytes were studied. The results obtained allowed us to: establish the mechanism that triggers anoxic crises; test a monitoring system for evaluating some parameters able to emphasize risks for the environment; avoid anoxic crises by suggesting practical actions to the Local Administration.

Female lung cancer mortality and long-term exposure to particulate matter in Italy

Background: Outdoor air pollution and particulate matter (PM) have recently been classified in Group 1 by IARC. In Italy there is no epidemiological study on the association between female lung cancer and PM as measured by the official monitoring stations. Methods: We estimated the dose–response relationship between female lung cancer mortality and available long-term outdoor PM10 and/or PM2.5 concentrations for all the Italian province capital city municipalities (respectively, 64 and 32 municipalities). Multiple regression analysis of standardized mortality rates (SMRates) for the period 2000–11, as a function of PM concentrations, considering percentage of smokers and deprivation index as additional explanatory variables, was performed for PM10 only. Results: The number of province capital cities with available PM2.5 data was not sufficient to detect a significant increment of SMRates as a function of concentrations. An SMRate increase of 0.325 for 1 &mgr;g m−3 increment of PM10 concentration was calculated. Moreover, the attributable risk of the overall SMRates for the two subgroups of municipalities under/equal and above 20 &mgr;g m−3 value was evaluated. Attributable deaths were computed by both the unitary SMRate increase and the attributable risk. A rough estimate of the impact of PM10 exposure at level above the WHO guideline value of 20 &mgr;g m−3 in these 64 municipalities is between 2920 and 3449 lung cancer deaths out of 22 162 (13–16%). Conclusion: Maintaining the PM10 concentrations below such WHO recommendation, an overall saving of nearly 300 lung cancer deaths per year in a population of 8 146 520 women living in the municipalities at study has been evaluated.