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The impact of municipal solid waste incineration (MSWI) on the environment and on human health was assessed by a life cycle assessment (LCA) approach. Even if risk assessment and epidemiologic analyses are specifically indicated for the investigation of the health outcomes, they resulted costly, time intensive and generally focused only on the effects caused by pollutant compounds directly emitted by the facility. Differently, LCA approaches are less time and cost intensive and able to account also for other indirect and direct emission of MSWI. However, results returned by LCA are based on average pollutant diffusion and individual exposure models limiting their representativeness for the specific context investigated. Furthermore, LCA is not able to return information about the final health outcomes caused by the pollutants emitted. The LCA performed for the Italian MSWI detected avoided impacts of about -0.11 kgPM 2,5eq /tonne of MSW and of about -2.5 × 10 -3  kgSb eq /tonne MSW for particulate matters and resource depletion, respectively. Positive impacts of about 900 kgCO 2eq /tonne MSW and about 15,000 CTUe/tonne MSW were detected for global warming and freshwater ecotoxicity indicators, respectively. Avoided impacts of about -1 × 10 -6  CTUh/tonne MSW and of about -2 × 10 -4  DALY/tonne MSW were also detected for human toxicity cancer and human health indicators, respectively. Epidemiologic studies referred to different Italian and EU reported some correlations among MSWI and some specific cancer and non-cancer health outcomes. By the way, these resulted affected by some methodological limitation preventing the definitive identification of causal nexus. In any case, a general coherence between LCA and epidemiologic approaches was detected. Furthermore, a particular correspondence was found between LCA results and biomonitoring studies concerning the concentration of heavy metals in blood and urinary samples of exposed individuals. All this highlighted the important role that LCA can have in supporting health impact assessment of MSWI in combination with epidemiologic approaches., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: In 2014, the European Environment Agency estimated 59 630 premature deaths in Italy attributable to long-term exposure to PM2.5, 17 290 to NO2 and 2900 to O3. The aim of this study was to test an approach for assessing health impact of the above pollutants analyzing possible associations between annual municipal concentrations, estimated by the national dispersion model developed by ENEA, and mortality rates for trachea, bronchus and lung (TBL) cancer, total respiratory diseases (RD) and chronic obstructive pulmonary diseases (COPD). Tuscany was selected as test case., Methods: For the 287 municipalities, 2009-13 standardized mortality rates (SMRates) for each cause of death were calculated by the ENEA epidemiological database. The SMRates of municipalities, aggregated on the basis of the 2003 or 2010 estimated pollutant concentration tertiles, were also computed., Results: TBL cancer SMRate in municipalities with 2003 PM2.5 levels >15.2 μg/m3 was significantly higher than the SMRates of the two lowest tertiles and COPD SMRates in the two highest O3 tertiles were significantly higher than that of the lower tertile. No association between PM2.5 or NO2 concentrations and RD and COPD was detected. Approximately 625 TBL cancer deaths attributable to PM2.5 levels above 10 μg/m3 in 2003 were estimated in the region. Smoking habits and deprivation index were homogeneously distributed among municipalities., Conclusion: This methodological approach allowed detecting associations between mortality and specific air pollutants even at levels below the Italian normative limits and could be employed to evaluate the potential health impact of air pollution in areas where direct measures of concentration are unavailable., (© The Author(s) 2018. Published by Oxford University Press on behalf of the European Public Health Association. All rights reserved.)

An assessment of potential carcinogenic and toxic health outcomes related to atmospheric emissions from the new-generation coal fired power plant of Torrevaldaliga Nord, in Central Italy, has been conducted. A chemical-transport model was applied on the reference year 2010 in the area of the plant, in order to calculate airborne concentrations of a set of 17 emitted pollutants of health concern. Inhalation cancer risks and hazard quotients, for each pollutant and for each target organ impacted via the inhalation pathway, were calculated and mapped on the study domain for the overall ambient concentrations and for the sole contribution of the plant to airborne concentrations, allowing to assess the relative contribution of the power plant to the risk from all sources. Cancer risks, cumulated on all pollutants, resulted around 5 × 10 -5 for the concentrations from all sources and below 3 × 10 -7 for the plant contribution, mainly targeting the respiratory system. On each part of the study domain, the plant contributed for less than 6% to the overall cancer risk. Hazard quotients from all sources, cumulated on all pollutants, reached values of 2.5 for the respiratory and 1.5 for the cardiovascular systems. Hazard quotients of non-carcinogenic risks from the plant, cumulated on all pollutants, resulted below 0.03 for the respiratory system and 0.02 for the cardiovascular system. On each part of the study domain, the plant contributed for less than 5% to the respiratory and cardiovascular risks. Both cancer risks and hazard quotients related to the plant are far below international thresholds for human health protection, while the values from all sources require consideration. The proposed method provides an instrument for prospective health risk assessment of large industrial sources, with some limitations presented and discussed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Perfluoroalkyl substances (PFAS), a heterogeneous group of highly stable man-made chemicals, have been widely used since 1960s and can be detected almost ubiquitously in all environmental matrices. In Italy, on January 2014, drinking water contamination in an area of the Veneto Region was detected mainly due to the drain of fluorinated chemicals by a manufacturing company operating since 1964., Methods: The present ecological mortality study was aimed at comparing mortality for some causes of death selected on the basis of previous reported associations, during the period 1980-2013, in municipalities with PFAS contaminated and uncontaminated drinking water on the basis of the levels indicated by the Italian National Health Institute (ISS). Sex-specific number, standardized mortality rates and rate ratios (RR) for PFAS contaminated and uncontaminated areas were computed for each cause of death through the ENEA epidemiological database., Results: In both sexes, statistically significant RRs were detected for all causes mortality, diabetes, cerebrovascular diseases, myocardial infarction and Alzheimer's disease. In females, RRs significantly higher than 1.0 were also observed for kidney and breast cancer, and Parkinson's disease. Increased risk, although not statistically significant, was observed for bladder cancer in both sexes, and for testicular cancer, pancreatic cancer and leukemia in males only., Conclusions: Higher mortality levels for some causes of death, possibly associated with PFAS exposure, were detected in contaminated municipalities in comparison with uncontaminated ones with similar socioeconomic status and smoking habits. These results warrant further individual level analytic studies to delineate casual associations., (© The Author 2017. Published by Oxford University Press on behalf of the European Public Health Association. All rights reserved.)

Persistent organic pollutants (POPs), such as PCBs (polychlorinated biphenyls) and DDT [1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane], are environmental contaminants with potential endocrine disrupting activity. DNA methylation levels in peripheral blood lymphocytes have been associated with serum concentrations of POPs in Greenland Inuit and Korean populations. Greenland Inuits are characterized by the highest worldwide POP levels. In this cross-sectional study we evaluated the relationship between serum POP concentrations and DNA methylation levels in sperm of non-occupationally exposed fertile men from Greenland, Warsaw (Poland), and Kharkiv (Ukraine). Serum levels of PCB-153 [1,2,4-trichloro-5-(2,4,5-trichlorophenyl)benzene], as a proxy of the total PCBs body burden, and of p,p'-DDE [1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene], the main metabolite of DDT were measured. Sperm DNA methylation level was assessed globally by flow cytometric (FCM) immunodetection of 5-methyl-cytosines and at specific repetitive DNA sequences (Alu, LINE-1, Satα) by PCR-pyrosequencing after bisulfite conversion. Multivariate linear regression analysis was applied to investigate correlations between serum POP concentrations and DNA methylation. No consistent associations between exposure to POPs and sperm DNA methylation at repetitive DNA sequences were detected. A statistically significant global decrease in methylation was associated with exposure to either POP by FCM analysis. This is the first study to investigate environmental exposure to POPs and DNA methylation levels considering sperm as the target cells. Although POP exposure appears to have a limited negative impact on sperm DNA methylation levels in adult males, the global hypomethylation detected by one of the methods applied suggests that further investigation is warranted., (© 2016 Wiley Periodicals, Inc.)

Background: the large amount of asbestos used in many Countries (including Italy) is causing an epidemic of asbestos related diseases, which is still ongoing because of their long latency., Objectives: this study is aimed at reconstructing Italian time series of deaths for mesothelioma in the period 1970-2009 and comparing Italian incidence and mortality data., Design, Setting, and Participants: deaths for pleural cancer (1970-2003,2006-2009) and mesothelioma (2003, 2006-2009) were recorded by the Italian Institute of Statistics (Istat) and provided by the Italian National Agency for New Technologies, Energy and the Environment (ENEA), incidence cases (1993-2008) were provided by the Italian mesotheliomas register (ReNaM) at the Italian National Workers' Compensation Authority (Inail). For the period before ICD-10 implementation (1970-2002) and when Istat data (2004-2005) are lacking, mesothelioma deaths were estimated through statistical models (logistic, Poisson). National incidence and mortality data were compared during the overlapping period (2003, 2006-2008)., Results: the mortality curve strongly rises from 1970 and seems to be smoothed in the last years. Mortality caused by mesothelioma and incident cases with certain diagnosis are overlapping, as are mortality due to pleural cancer other than mesothelioma and mesothelioma incidence with uncertain diagnosis (probable/possible)., Conclusions: this epidemiological analysis of deaths encoded as pleural tumour suggests to carefully investigate space-temporal distribution before excluding they could be mesotheliomas. Some new lights have been thrown on the statistical behaviour of mesothelioma mortality.

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., (© 2014 Wiley Periodicals, Inc.)

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.

Purpose: Past intensive use of asbestos has implied severe public health consequences. Spatial distribution of deaths from malignant mesothelioma and of compensated cases for asbestos related diseases in Italy were compared to identify unexpected sources of asbestos exposure., Methods: Mortality for malignant mesothelioma at municipal level and geographical clusters of compensated cases for asbestos related diseases, as proxy of industrial asbestos exposure, were identified in the period 1988-2001., Results: Municipalities with at least four mesothelioma deaths and a statistically significant mortality excess were 148; and 53 out of them had no compensated case for asbestos-related diseases. Finally 22 of these lay outside of any aforementioned cluster, thus suggestive of a possible unrecognized exposure., Conclusions: Availability long-term national figures and the different etiology of asbestos related diseases are the key features of this exercise that was applied to Italy, but can be replicated wherever registration systems of diseases related to long term exposure to asbestos are available.

Background: Occupational exposure to asbestos has been widely reported in the Region, but a high risk for non-occupational and environmental contaminations have also been documented., Objectives: To describe the geographical distribution ofpleural cancer deaths and compensated asbestosis cases from 1980 to 2001 in the Lazio Region., Methods: For each municipality Standardized Mortality Ratios (SMRs) for pleural cancer and Standardized Incidence Ratios (SIRs) for asbestosis were estimated. Expected cases were estimated from age and gender specific rates in Lazio. SatScan software was used to identify clusters and to verf;j their statistical significance., Results: 789 deaths from pleural cancer (495 males and 294 females) occurred in Lazio from 1980 to 2001. The standardized mortality rate per 100.000 inhabitants is 0,74 (0,95 for males and 0,54 for females). The main excess mortality from pleural cancer occurred in the municipalities of Civitavecchia (SMR: 269,9; 95% CI: 164,9 - 416,8), Colleferro (SMR: 304,9; 95% CI: 139,4-578,8) and Rocca Priora (SMR: 379,2; 95% CI: 103,3-970,9). Significant SIRs for compensated asbestosis cases were found in the industrial areas of the Naples-Rome highway and in the shipyard area of Civitavecchia. Nofemale compensated cases were found. The most important clusters were identified in the municipality of Civitavecchia for pleural cancer (p-value = 0,117) and in the Colleferro industrial area for compensated asbestosis cases (p-value = 0,001)., Conclusions: Epidemiological surveillance of incident cases of malignant mesothelioma in the Lazio Region and the investigation of modalities of asbestos exposure are urgently needed for prevention of occupational diseases.

Amyotrophic lateral sclerosis (ALS) is a rare and devastating neurological disorder of the adult age with a prognosis of about 2-3 years from the onset of the disease. No clear cause has been identified but it seems to be a multifactorial disease with genetic and environmental components involved. Increments of mortality rates were observed since 1980 both in Italy and in many other countries. The objective of the present study is to describe the distribution of ALS mortality in Italy in the period 1980-2001 detecting single municipalities or clusters with high mortality levels for motor neuron disease (MND). ALS represents the main part (85%) of the MND group which is globally identified by the IX ICD (International Classification of Diseases and Causes of Death) 335.2 code. Death numbers and standardized mortality ratios (SMR) for MND were calculated for all Italian municipalities through the ENEA mortality database system (data source: National Institute of Statistics-ISTAT), using national mortality rates as reference. Subsequently, in order to detect municipal clusters, spatial analysis was performed. Out of the 8,099 Italian municipalities, 132 where characterized by SMR values higher than expected. Moreover 16 clusters with significant high relative risk values (RR) were identified, 12 out of them including only a single municipality. Only 22 of the municipalities with high SMR were included in the clusters. In conclusion, the two different epidemiological methodologies demonstrated to be widely complementary in detecting the geographical distribution of the disease in terms of risk for populations. A first selection of the priority areas where analytical studies should be carried on, in order to identify risk factors associated to ALS, is tentatively suggested.

Objective: To describe the mortality rates for motor neuron disease (MND) in Italy both at national level and at three large geographical sub-areas (Northern, Central, Southern Italy)., Design: Deaths for MND are coded accordingly to ICD (IX revision) with 335.2 code. Mortality for MND is analysed for the period 1980-99. Mortality rates are age-standardized on the structure of the 1991 Italian population. Mortality data are derived from the National Institute for Statistics (ISTAT) and are made available by the Epidemiological Database of the Italian National Agency for New Technologies, Energy and the Environment (ENEA)., Main Outcome Measures: Age adjusted mortality rates for MND are calculated for 5-year periods (1980-84, 1985-89, 1990-94, 1995-99) at both national level and three geographical sub-areas (north, center and south); sex and age specific mortality rates are also reported for two decades (1980-89 and 1990-99)., Results: During 1980-99 the annual age-standardized mortality rate in Italy was 1.35 x 100000 in males and 1.10 (x 100000) in females. In the period 1995-99 the mortality rates increased by 39.3% in males and 78.2% in females at national level when compared to 1980-84 rates (1.56 vs 1.12 deaths per 100000 for males and 1.39 vs 0.78 per 100000 for females in 1980-84 and 1995-99, respectively). In the three large geographical sub-areas such increases were 37.6%, 29.7% and 57% in males and 73%, 63.1% and 114.3% in females, respectively., Conclusion: The increase reported in this study is probably due to a mix of different factors as population ageing (age-specific rates reach a peak in the age class 70-74 years), better accuracy of death certificates, adoption of new clinical criteria and at last a wide spread of environmental risk factors (metals, solvents, pesticides, electromagnetic fields) and modification of life style (smoking, diet, professional sport).

Objective: To his study describes the geographical distribution of pleural cancer deaths and asbestosis cases from 1980 to 2000 in Sardinia Region (Italy). For both conditions regionwide registration systems have been available for a relatively long time and allow the identification of statistically significant clusters., Design: For each town we have estimated Standardized Mortality Ratios (SMRs) for pleural cancer and Standardized Incidence Ratios (SIRs) for asbestosis. Expected cases were estimated from age- and gender specific rates in Sardinia. SatScan software was used to identify clusters and to verify their statistical significance., Setting: Sardinia Region (Italy)., Main Outcome Measures: Standardized mortality and incidence rates respectively for pleural cancers and asbestosis cases and territorial clusters., Results: The most important cluster of pleural cancer was identified in the area defined by Carloforte, Calasetta, Portoscuso and Sant'Antioco municipalities (Southwestern Sardinia) with 15 observed cases (p value= 0.003). Other clusters were detected in the municipality of La Maddalena (11 observed cases against 1.91, expected p value= 0.008) and in Southern Sardinia between Cagliari and Sarroch (p value= 0.018). The town of Marrubiu is clearly the most important cluster (p value= 0. 001) with 6 asbestosis cases in the period., Conclusions: These results indicate the urgency of the epidemiological surveillance of asbestos related diseases in Sardinia. The active search for incident cases of malignant mesothelioma in the whole Region and the analysis of modalities of asbestos exposure (according to national guidelines) is an indispensable tool for the primary prevention of occupational, environmental and domestic exposures from unknown asbestos sources of contamination.

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., ((c) 2004 Wiley-Liss, Inc.)

Objective: To investigate cause-specific mortality in an area of Campania region, in the surroundings of Naples, characterized by many toxic waste dumping grounds sites and by widespread burning of urban wastes., Design: The study area was characterized by examining the spatial distribution of waste disposal sites and toxic waste dumping grounds, using a geographic information system (GIS). Mortality (1986-2000) was studied in the three municipalities of Giugliano in Campania, Qualiano and Villaricca, encompassing a population of about 150,000 inhabitants. Mortality rates of the population resident in the Campania region were used in order to generate expected figures., Outcomes: Causes of death of a priori interest where those previously associated to residence in the neighbourhood of (toxic) waste sites, including lung cancer, bladder cancer, leukemia and liver cancer., Results: Overall 39 waste sites, 27 of which characterized by the likely presence of toxic wastes, were identified in the area of interest. A good agreement was found between two independent surveys of the Regional Environmental Protection Agency and of the environmentalist association Legambiente. Cancer mortality was significantly increased, with special reference to malignant neoplasm of lung, pleura, larynx, bladder, liver and brain. Circulatory diseases were also significantly in excess and diabetes showed some increases., Conclusions: Mortality statistics provide preliminary evidence of the disease load in the area. Mapping waste dumping grounds provides information for defining high risk areas. Improvements in exposure assessment together with the use of a range of health data (hospital discharge cards, malformation notifications, observations of general practitioners) will contribute to second generation studies aimed at inferring causal relationships.

Objectives: The purpose of the present study was to investigate the association between mortality from chronic obstructive pulmonary disease (COPD) and exposure to fluoro-edenite, a newly discovered amphibolic fiber found in Biancavilla (Sicily), a municipality on the slope of the Etna volcano, where a high mortality from malignant mesothelioma had been previously observed., Methods: Thirty-six municipalities located in the volcanic area of mount Etna were selected for study. An ecological regression model was applied with mortality from COPD as the dependent variable, mortality from mesothelioma as a proxy for exposure to fluoro-edenite, and lung cancer mortality, an urban-rural index, a deprivation index and an aging index as the predictors of COPD mortality. For each municipality, risk classes were determined for the continuous variables with the use of a finite mixture model., Results: A significant association was found between COPD mortality and pleural neoplasm mortality among the women in this study. The association was less consistent for the men., Conclusions: Despite the limitations of this ecological study, the results cohere with the results of toxicologic and observational studies and suggest an etiologic role for fluoro-edenite in nonmalignant respiratory diseases.

By assessing mortality causes, the authors have described the health status of people living in 3 areas of Tuscany around Livorno, Orbetello and Piombino municipalities. Direct standard mortality and local standard mortality ratios were assessed as regards the years 1988?1997. Total mortality turned out to be equal or lower than the Italian average, with the exception of the women from Livorno municipality. Cancer mortality is higher in Livorno area, among females in Orbetello area and males in both Orbetello and Piombino municipalities. As regards regional mortality, the situation in Livorno seems to be the worst, followed by Orbetello and Piombino.

The aim of the present work was to compare mortality data during 1980-94 (24 causes of death for males and 25 for females) in populations living in municipalities with different urbanization levels of seven Italian regions (Piemonte, Lombardia, Emilia Romagna, Marche, Lazio, Campania and Sicilia). Urbanization levels were established following the classification of the Italian Institute of Statistics (ISTAT, Classificazione dei comuni secondo le caratteristiche urbane e rurali, 1986). A quite uniform configuration of causes of death with characteristic excesses in the urban environment was evident. The only exception was represented by the male population in Lombardia, probably due to the prevalence of occupational exposure. The urban excesses included: all tumors (also for the population < 64 year old), trachea, bronchi and lung tumors (also < 64 year old people), colorectal cancer, bladder cancer, male liver cancer and female breast cancer. Mortality due to stomach cancer and, only in males, to larynx cancer was lower in urban than in the other types areas, except the Southern regions. Concerning some non tumoral pathologies a more dyshomogeneous situation was observed, with a prevalence of urban defects in Piemonte and Lombardia, of urban excesses in the Southern regions and of both excesses and defects in Emilia Romagna and the Central regions. The "semi-rural" and "rural" types municipalities resulted quite similar, with the exception of Campania and Lazio. In fact, in these two regions a greater association with stomach cancer and lower mortality for all other tumors compared to the urban and, at a lesser extent, to the semi-urban municipalities was found. However, the lower tumor mortality was partly balanced by some other causes of death, again with the exception of Campania and at a lesser extent of Sicilia. On the basis of the causes of death comparisons, the semi-urban municipalities seem to represent a transition type between the urban and the rural ones.

L-acetylcarnitine plays a key role in sperm metabolism and in the whole spermatogenetic process. In the present work, the influence of L-acetylcarnitine, administered i.p. (100 mg/kg body weight), on the recovery processes of mouse spermatogenesis after local acute irradiation with 10 Gy X-rays has been investigated. The effects were monitored 28, 35, 40, 45, 50, 55, and 60 days after irradiation by flow cytometric analysis of cellular DNA content. In the LAC-treated animals, the fraction of tetraploid cells is higher at 28 (p less than 0.05) and 45 days (p less than 0.02). Corresponding with the timing of the stages of murine spermatogenesis, the round spermatid fraction is higher at 45 days (p less than 0.1) and the elongated spermatid fraction is higher at 50 days (p less than 0.1) after irradiation. In addition, the LAC-treated animals show a faster recovery throughout the maturation process, from tetraploid to round and elongated spermatids. These results indicate that the presence of exogenous LAC could enhance the recovery of spermatogonial cells.

Some 150 tumor specimens from 49 patients with non-small-cell carcinoma of the lung (23 epidermoid, 14 adenocarcinoma, 12 large-cell carcinoma) and three with nonneoplastic lung disease were analysed for cellular DNA content by flow cytometry. Monodispersed cells were stained with ethidium bromide and mithramycin. Normal specimens and samples from patients with nonneoplastic disease constantly yielded a single cell population with diploid DNA content. Twenty of 23 epidermoid carcinomas exhibited one or more than one aneuploid subpopulation. Ten of 12 large-cell carcinomas were characterized by one aneuploid clone and 2/12 by two aneuploid clones. Adenocarcinoma exhibited multiclonal cell subpopulations (one to five aneuploid clones). Further information has been obtained on the differential presence of clones in various tumor areas and in infiltrated lymph nodes. These tumors appear characterized by a remarkable degree of cellular heterogeneity. The cytometric ploidy level(s) and the cell population multiclonal structure yield, in comparison with, and in addition to, pathology, indications of possible clinical interest. A correlation between the clonal DNA content and a prognostic parameter such as the tumor mass doubling time has been demonstrated.

The relative DNA content of cellular samples from 54 patients affected by breast carcinomas and 20 affected by benign breast lesions (including 11 fibroadenomas) was measured by flow cytometry. All normal tissue samples and 17/20 (85%) specimens from benign lesions exhibited a cytometrically diploid DNA distribution, 3/20 (15%) benign lesions an abnormal DNA content, and 35/54 (65%) carcinomas at least one aneuploid cell subpopulation. Furthermore, 9/54 (17%) tumors were characterized by the presence of more than one aneuploid cell subpopulation. The results also indicate that flow cytometry can be used to recognize lymph nodes infiltrated by aneuploid cells. Statistically significant correlations were evidenced between the occurrence of aneuploidy or the ploidy level measured as DNA index and the nodal infiltration status. The percentage of S cells can also be extracted from DNA content distribution histograms. Statistically significant differences (p less than 0.01) were also observed for the percentage of S cells between normal tissues (6.2 +/- 3.2 SD) and benign lesions (11.1 +/- 6.6 SD), normal tissues (6.2 +/- 3.2 SD) and aneuploid tumors (19.7 +/- 10.3 SD), benign lesions (11.1 +/- 6.6 SD) and aneuploid tumors (19.7 +/- 10.3 SD), and diploid (7.9 +/- 4.0 SD) and aneuploid tumors (19.7 +/- 10.3 SD).

An automatic procedure for recovering the DNA content distribution of mouse irradiated testis cells from flow cytometric histograms is presented. First, a suitable mathematical model is developed, to represent the pattern of DNA content and fluorescence distribution in the sample. Then a parameter estimation procedure, based on the maximum likelihood approach, is constructed by means of an optimization technique. This procedure has been applied to a set of DNA histograms relative to different doses of 0.4-MeV neutrons and to different time intervals after irradiation. In each case, a good agreement between the measured histograms and the corresponding fits has been obtained. The results indicate that the proposed method for the quantitative analysis of germ cell DNA histograms can be usefully applied to the study of the cytotoxic and mutagenic action of agents of toxicological interest such as ionizing radiations.