In vitro and in vivo mutagenicity studies with airborne particulate extracts

The contribution of nitro compounds to airborne particulate mutagenicity was studied with Salmonella typhimurium strains TA98, TA98NR, TA98/1,8DNP6. The results obtained indicate that nitropyrenes play a minor role in air particulate mutagenicity. Seasonal variations indicate a relatively greater contribution of nitro compounds to the mutagenicity of spring and summer samples. Fractionation of extracts into acidic, neutral and basic components shows that neutral compounds account for about two-thirds of the total mutagenic activity. Attempts to extract mutagens adsorbed onto particulate matter with aqueous media were almost completely negative. No significant mutagenicity was detected in urine and faecal extracts and in plasma samples of Sprague-Dawley rats treated with air particulate extracts at 80 mg/kg either per os or by i.p. injection. Negative results were obtained in the micronucleus test with Swiss mice treated at 200 and 400 mg/kg (twice by i.p. injection). A significant decrease in liver aminopyrine-N-demethylase was observed in Swiss mice injected with air particulate extracts or its basic and neutral fractions. In vitro experiments suggest a direct interaction of test materials with microsomal cytochrome P-450.     

Mutagenic activity of airborne particulate matter in a petrochemical industrial area

Exposure to airborne particulate matter has adverse effects on human health and ecosystem. Mutagenic activity of airborne particulate organic matter extracts in three time periods from total suspended particles (TSP) and particles less than 10 microm (PM10) was evaluated in an area under the influence of a petrochemical industry located in the town of Triunfo, Brazil. The extracts were investigated using the Salmonella/microsome assay, with the microsuspension method. The extracts were obtained by sonication extracted using dichloromethane (DCM) solvent. The fractions were tested for mutagenicity with the Salmonella typhimurium strains TA98 (with and without metabolic activation), TA98NR and TA98/1,8DNP(6); or YG1021 and YG1024. A positive frameshift mutagenic response was observed for the environmental samples during the different periods. The responses according to percentage of extractable organic matter (EOM%), EOM/m(3), revertants/microg (rev/microg) and revertants/m(3) (rev/m(3)) were lower for TSP than for PM10 extracts. The highest rev/m(3) values were observed in PM10 extract samples collected in winter, July 2005, in the presence (13.79 rev/m(3)) or absence (6.87 rev/m(3)) of S9 fraction. Similarly in the first (1995) or second period (2000) the highest values for TSP were observed in winter, but with lower activity (3.00 and 0.89 rev/m(3) respectively). The responses observed for the nitrosensitive strains suggest the contribution of nitro, amino and/or hydroxylamino derivatives of PAHs to the total mutagenicity of matter extracted from airborne particles. The Salmonella/microsome assay was a sensitive method to define areas contaminated by genotoxic compounds, even in samples with TSP or PM10 values that are acceptable according to legal environmental quality standards, favoring environmental control measures with an effective response seen in the population's improved quality of life.     

Cytogenetic evaluation of extractable agents from airborne particulate matter generated in the city of Catania (Italy)

In order to document cytogenetic damage associated with air pollution and, possibly, with health effects in the city of Catania, Sicily (Italy), we analyzed the induction of chromosomal aberrations by extractable agents from airborne particulate matter in a Chinese hamster epithelial liver (CHEL) cells. These cells retain their metabolic competence to activate different classes of promutagens/procarcinogens into biologically active metabolites. Airborne particulate matter was obtained from two stationary samplers (stations I and II) in two areas endowed by an elevated car transit in the centre of Catania. The results obtained clearly indicated that airborne particulate matter from both stations I and II proved to be clastogens in CHEL cells but not in Chinese hamster ovary (CHO) cells without metabolic activation, indicating that airborne particulate mixtures need to be metabolically converted before exerting their genotoxic potential. On the basis of these results we can assert that the test system employed to identify the cytogenetic potential of airborne particulate matter is useful and profitable for environmental control, and helpful to plan specific actions aimed at reducing the hazards derived from exposure to polluted air.     

Measuring personal exposure to airborne mutagens and nicotine in environmental tobacco smoke

The exposure of individuals to environmental tobacco smoke (ETS) is of increasing public health concern because epidemiological studies have associated passive smoking with increased risk of a variety of adverse health effects among non-smokers including lung cancer. As a way to measure individual exposure to the mutagenic compounds in the complex mixture of ETS, we used a sensitive Salmonella/microsome micro pre-incubation (microsuspension) assay to detect mutagenicity of particulate matter collected on filters from low volume (1.7 1/min flow rate) personal sampling pumps. Airborne nicotine was collected concurrently as a marker for ETS exposure. In pilot-field studies, individual exposure to ETS was measured in two separate indoor environments in which smokers were present: a gambling casino and a bingo parlor. Total suspended particulate matter (TSP) was collected on filters worn near the breathing zone of non-smoking individuals. Sampling times ranged from 40 min to 6 h. All extracts of filters had detectable levels of mutagenic activity (TA98, +S9) resulting in airborne mutagenic activity concentrations of 500-5000 rev/m3. The mutagenic activity of the filters from the casino and bingo parlors was significantly correlated with total particulate matter per filters (n = 12; Rho = 0.85, p less than 0.01) and with airborne nicotine per filter (n = 12; Rho = 0.95, p less than 0.01). The microsuspension assay was sufficiently sensitive to detect the mutagens associated with extracts of particulate matter from low volume samples (0.2-0.6 m3) in these indoor environments over a relatively short sampling time, and could be useful in studies of personal exposure to the mutagens in environmental tobacco smoke. Further, airborne nicotine concentrations were highly correlated with airborne mutagenicity and the mutagenic activity associated with ETS could therefore be estimated by the concentrations of nicotine.     

DNA adduct formation from the mutagenic air pollutant 3-nitrobenzanthrone

The environmental contaminant 3-nitrobenzanthrone (3-nitro-7H-benz[d, e]anthracen-7-one) was recently shown to be a very strong bacterial mutagen, suggesting a new class of mutagenic compounds present in airborne particulate matter and diesel exhaust. Using the 32P-postlabeling assay, we investigated the capacity for 3-nitrobenzanthrone to form DNA adducts in vitro. Calf thymus DNA was incubated with 3-nitrobenzanthrone and either xanthine oxidase, a mammalian nitroreductase or rat liver S9 or zinc. Under these conditions 3-nitrobenzanthrone formed a total of seven adducts detectable by 32P-postlabeling. Using enrichment by butanol extraction the highest level of DNA adduct formation was found with activation by zinc (RAL: 88.4+/-32 per 108 nucleotides) followed by activation with xanthine oxidase (RAL: 75.5+/-12) and activation by rat liver S9 (RAL: 48.6+/-8). Three of the seven adduct spots were detected in all activation systems, however different amounts of individual spots were obtained in the different in vitro systems. The adduct pattern observed for the enzymatic incubations consisted of three major spots and was essentially identical. Chemical reduction of 3-nitrobenzanthrone by zinc resulted in five adduct spots whose formation was found to be concentration dependent. All adducts of 3-nitrobenzanthrone observed in this study migrated primarily along a diagonal zone, typical for DNA adducts derived from extracts of airborne particulate matter. When butanol enrichment was compared with nuclease P1 enrichment one adduct was clearly sensitive to the 3'-monophosphatase activity of nuclease P1. Our results demonstrate that 3-nitrobenzanthrone binds covalently to DNA after metabolic activation, forming multiple DNA adducts in vitro all of which are reduction products. These adducts may contribute to the known genotoxicity and carcinogenicity of extracts from airborne particulates.     

Chemical characterization and biological properties of airborne particulate matter

Summary The presence of heavy metals and Polynuclear Aromatic Hydrocarbons (PAH) has been evidenced in airborne particulate matters samples collected in the atmosphere of a non-industrialized middle size Italian town (150.000 inhabitants). The PAH concentrations, higher in winter than in other seasons, was inversely correlated with some meteorological variables such as temperature (p<0.01) and wind speed (p<0.05). The positive correlation found (p<0.01) between lead and total PAH seems to indicate traffic as the most important source of pollution.All samples were mutagenic in the Ames test and the highest mutagenicity levels were obtained in winter samples. The positive correlation found between Benzo(a)Pyrene {B(a)P} concentration and the number of revertants/m³ air, suggests that B(a)P could be an index of mutagenic potential of the matrix considered.The airborne particulate matter extracts obtained by organic solvents were able to inhibit the phagocytosis of mice alveolar macrophages, giving further proof of the possibility of interaction between air-particulate chemical compounds and important biological functions.     

Characterization of testudine melanomacrophage linear,membrane extension processes—Cablepodia—By phase and atomic force microscopy

Summary Melanomacrophages (MMs) are a component of an internal, pigmented cell system in liver and splenic tissues of some fishes, anurans, and reptiles. The cells have been found in centers or aggregates in sinusoids and are associated with cells capable of producing a peptide cytokine and immunoglobulins. A unique cell extension process has been observed in turtle MMs placed into cell culture, and this process has been studied by light and atomic force microscopy. These structures, referred to as cablepodia, are uniquely straight, narrow, and unbranching and appear to originate from growth cones opposite lamellipodia. Cablepodia were found to connect with other turtle MMs and fibroblasts forming cell networks. Dividing fibroblasts to which a cablepodium attached ceased cell division. The observations collectively suggest that a principal reason for aggregations of MMs in internal organs of lower vertebrates in their ability to form interconnected networks of cell processes for trapping and processing of particulate matter, cells and infectious organisms and, possibly, for the communication of cell signals and transfer of intracellular materials.     

Monitoring airborne genotoxicants in the rubber industry using genotoxicity tests and chemical analyses

This research was designed to examine the presence of mutagenic/carcinogenic compounds in airborne pollutants in the rubber industry using an integrated chemical/biological approach. Inhalable airborne particulate matter (PM-10: <10 microm) was collected in four rubber factories using a high-volume sampler equipped with a cascade impactor for particle fractionation. The organic extracts of two different fractions (0.5-10 microm and <0.5 microm) were examined for mutagenicity with the Ames test and for in vitro DNA-damaging activity in human leukocytes by single-cell microgel electrophoresis (Comet assay). The extracts were also studied by gas chromatography/mass spectrometry (GC/MS) for polycyclic aromatic hydrocarbon (PAH) content. Nitrosamines in ambient air were sampled on cartridges and analysed by GC with a thermal energy analyser (TEA) detector. Airborne volatile genotoxins were monitored in situ using a clastogenicity plant test (Tradescantia/micronuclei test). The results showed that airborne particulates were mainly very fine (<0.5 microm) and that trace amounts of genotoxic nitrosamines (N-nitrosodimethylamine: 0.10-0.98 microg/m(3); N-nitrosomorpholine: 0.77-2.40 microg/m(3)) and PAH (total PAH: 0.34-11.35 microg/m(3)) were present in air samples. Some extracts, particularly those obtained from the finest fractions, were mutagenic with the Ames test and genotoxic with the Comet assay. In situ monitoring of volatile mutagens using the Tradescantia/micronuclei test gave positive results in two working environments. The results showed the applicability of this integrated chemical-biological approach for detecting volatile and non-volatile genotoxins and for monitoring genotoxic hazards in the rubber industry.     

Mutagenic activity of airborne particulate matter in a petrochemical industrial area

Exposure to airborne particulate matter has adverse effects on human health and ecosystem. Mutagenic activity of airborne particulate organic matter extracts in three time periods from total suspended particles (TSP) and particles less than 10 μm (PM10) was evaluated in an area under the influence of a petrochemical industry located in the town of Triunfo, Brazil. The extracts were investigated using the Salmonella/microsome assay, with the microsuspension method. The extracts were obtained by sonication extracted using dichloromethane (DCM) solvent. The fractions were tested for mutagenicity with the Salmonella typhimurium strains TA98 (with and without metabolic activation), TA98NR and TA98/1,8DNP₆; or YG1021 and YG1024. A positive frameshift mutagenic response was observed for the environmental samples during the different periods. The responses according to percentage of extractable organic matter (EOM%), EOM/m³, revertants/μg (rev/μg) and revertants/m³ (rev/m³) were lower for TSP than for PM10 extracts. The highest rev/m³ values were observed in PM10 extract samples collected in winter, July 2005, in the presence (13.79 rev/m³) or absence (6.87 rev/m³) of S9 fraction. Similarly in the first (1995) or second period (2000) the highest values for TSP were observed in winter, but with lower activity (3.00 and 0.89 rev/m³ respectively). The responses observed for the nitrosensitive strains suggest the contribution of nitro, amino and/or hydroxylamino derivatives of PAHs to the total mutagenicity of matter extracted from airborne particles. The Salmonella/microsome assay was a sensitive method to define areas contaminated by genotoxic compounds, even in samples with TSP or PM10 values that are acceptable according to legal environmental quality standards, favoring environmental control measures with an effective response seen in the population's improved quality of life.     

施用有机肥对黑土团聚体有机碳的影响

采用湿筛法对不同处理下的黑土进行分离, 得到4种不同大小（>2 000、2 000～250、250～53、<53 μm）的团聚体,利用密度分组法分离出游离的轻组（free LF）和团聚体内的颗粒有机质（iPOM）,以评价有机肥对黑土团聚体有机碳的影响.结果表明：与对照相比,施用有机肥显著增加了土壤的团聚化作用,可在一定程度上抵消耕作对团聚体的破坏作用,减缓团聚体的周转.团聚体内细颗粒有机碳（fine iPOM-C）含量显著高于粗颗粒有机碳(coarse iPOM-C)含量,说明施用有机肥有利于团聚体内细颗粒有机碳的积累,是施用有机肥条件下黑土团聚体内碳固定的主要形式.     

Collaborative study using the preincubation Salmonella typhimurium mutation assay for airborne particulate matter in Japan. A trial to minimize interlaboratory variation.

A collaborative study has been performed over a period of 3 years to develop a suitable method for monitoring the mutagenicity of airborne particulate matter. The study was organized with 8 laboratories and performed in the following steps: (1) selection of a suitable technique for each process involved in the mutagenicity monitoring, (2) developing a tentative protocol by combining systematically the selected techniques, (3) evaluation of the protocol by intra- and inter-laboratory studies, (4) modification of the protocol according to the evaluation, and (5) evaluation of the modified protocol by conducting an interlaboratory study. We found a suitable method for mutagenicity monitoring of particles in the atmosphere. Airborne particles were sampled with a high-volume sampler, the samples were stored at -80 degrees C, extracted by sonication using dichloromethane, solvent-exchanged, and assayed by the preincubation method using Salmonella typhimurium TA98 and TA100. The observed mutagenic activity was normalized with that of an internal standard. Round robin tests revealed that the method resulted in excellent reproducibility. The coefficient of variation for mutagenic activities of airborne particulate samples collected in various districts of Japan were in the range of 14.7 +/- 6.6% to 19.6 +/- 4.0% for strains TA98 and TA100 with and without metabolic activation. We also found that the plate incorporation method was equivalent to the preincubation method for airborne particulate extracts.     

Analysis and tracing of polycyclic aromatic hydrocarbons and mutagenicity of airborne particulates from the Taipei area

In a 3-year study, we determined the mutagenicity and polycyclic aromatic hydrocarbons (PAHs) of airborne particulates collected during December 1987-September 1988 (216 samples), October 1988-January 1989 (81 samples), and October 1989-April 1990 (52 samples) from 9 locations in the Taipei area. We found that dichloromethane extracts of all the samples were mutagenic to Salmonella typhimurium in the Ames test. Moreover, the mutagenicity was much higher in the presence of rat liver microsomal fraction (S9 mixture) than that observed in its absence, which indicates that airborne particulates contained both direct and indirect mutagens. The average mutagenicity of the samples collected in the 3-year period was 137, 127, and 118 histidine revertants/10 m3 air, respectively. On the other hand, we found that dichloromethane extracts of each airborne particulate sample contained 14 PAHs with wide variations in concentration and relative distribution. The levels of Pha, Flu, Pyr, and Ben were much higher than the PAHs with higher ring numbers such as BaP, BeP, Pr, IP, and DbA. The average PAH content was 8.0, 5.0, and 7.8 ng/m3 air for airborne particulates collected during December 1986-September 1987, October 1988-January 1989, and October 1989-April 1990, respectively. Among the 9 stations, Fu Hsing Elementary School and Chung Hsing University (Taipei campus), which are, respectively, located in the downtown area and a heavy traffic zone, had significantly higher levels of mutagenicity and PAHs than did the other stations. Moreover, comparative analysis of PAH levels of airborne particulates over the 3-year period revealed an interesting season-dependent change of PAH content in airborne particulates from the Taipei area. The concentrations of individual and total PAHs were consistently lower in the summer than those in the winter. A similar pattern of seasonal change was also observed in the mutagenicity of airborne particulate samples examined. It is worth mentioning that neither PAH level nor mutagenicity of airborne particulates showed significant yearly change over the 3-year period of study. As part of an effort to identify pollution sources, we examined the mutagenicity and PAH compounds of air particulates collected from the burning of garbage (14 samples) and motor-vehicle exhaust in the Hsin Hai Tunnel (17 samples), Taipei. The results showed that garbage burning gave rise to air particulates containing several hundred times higher levels of PAHs and about 20 times stronger mutagenicity, while the motor-vehicle exhaust contained about ten times higher PAH content and mutagenicity as compared with those of airborne particulates of the Taipei city.(ABSTRACT TRUNCATED AT 400 WORDS)     

Polarized distribution of inducible nitric oxide synthase regulates activity in intestinal epithelial cells

Inducible nitric oxide synthase (iNOS) functions as a homodimer. In cell extracts, iNOS molecules partition both in cytosolic and particulate fractions, indicating that iNOS exists as soluble and membrane associated forms. In this study, iNOS features were investigated in human intestinal epithelial cells stimulated with cytokines and in duodenum from mice exposed to flagellin. Our experiments indicate that iNOS is mainly associated with the particulate fraction of cell extracts. Confocal microscopy showed a preferential localization of iNOS at the apical pole of intestinal epithelial cells. In particulate fractions, iNOS dimers were more abundant than in the cytosolic fraction. Similar observations were seen in mouse duodenum samples. These results suggest that, in epithelial cells, iNOS activity is regulated by localization-dependent processes.     

Inhibition of desmosome formation in chick cell aggregates.

Desmosomes (macula adherens) have been associated with the function of adhesion. Their possible role in aggregation and sorting of chick and mouse epithelial cells has been investigated. Treatment of aggregates with 2-5 microgram/ml of actinomycin D which inhibited RNA synthesis also inhibited both desmosome formation and aggregation if administered at the beginning of the aggregation process. In contrast, if the drug was administered at six hours, when the cells had recovered from the process of dissociation, then aggregation over the following six hours appeared normal from observation of living samples. Such aggregates incorporated leucine-3H at roughly 85% of the control level. A quantitative comparison was made of desmosome formation in aggregates treated with actinomycin D for hours 6-12 and those cultured in normal medium. Desmosome formation was inhibited by the drug, although aggregation could proceed. Combinations of chick corneal and mouse skin cells sorted out in the presence of actinomycin D to the same extent as controls. Thus desmosome formation, which normally occurs during aggregation of the epithelial cells studied here, is not coupled with the aggregation or cell sorting process in these cells of stratified epithelia. When cells were treated with cycloheximide (100 muM) both desmosome formation and the progressive rounding up of aggregates was inhibited.     

Inhibition of quantitative PCR analysis of fungal conidia associated with indoor air particulate matter

Environmental samples analyzed by quantitative PCR (qPCR) are subject to interference by inhibitors present in the environment being sampled. A controlled determination of the effect of inhibitors associated with sampling indoor air and the ability of internal standard controls to detect inhibition was carried out by filter collection of air samples followed by spiking of the filters with green fluorescent protein-expressing Aspergillus fumigatus conidia. Microscopic conidial counts were compared with qPCR results and correlated with levels of particulate matter and viable airborne microorganisms. Our data showed that PCR can be inhibited by masses of particulate matter as low as 50 μg and that the amount of inhibition was positively correlated with the mass of particulate (r = 0.75) and the number of non-filamentous organisms (r = 0.73). The use of internal standard DNA identified the presence of inhibitors and indicated the need for additional sample processing or qualification of sample results.     

Analysis of mutagenic activity of airborne particulate matter, standard reference materials and reference compounds using base pair-specific Salmonella typhimurium tester strains

The mutagenicity profiles of organic extracts of airborne dust samples from Mannheim, Germany, and two standard reference materials (SRM) as well as eight compounds with different chemical properties were investigated using tester strains Salmonella typhimurium TA700x (Ames II Assay). Each strain of this series carries a unique missense mutation in the histidine operon and is reverted by only one specific base substitution out of six possible changes. Mutation patterns of eight compounds with different modes of genotoxic action reveal significant differences. Samples of airborne particulate matter (APM) from an industrialized town in Germany (Mannheim) were collected for five consecutive days once a month for 1 year using an automatic high-volume air sampler. Samples taken from Monday to Friday were Soxhlet-extracted and prepared according to standard methods. Although the threshold limit for the least active strains is not triggered by all samples, it can be concluded that mutation patterns of the samples do not vary between different seasons. Standard reference materials (SRMs) were prepared and tested using the same methods. SRMs and APM samples from Mannheim reveal similar mutagenicity profiles in TA700x strains. The comparison of the mutagenicity profiles of air dust extracts from Mannheim and the SRMs, respectively, with reference compounds investigated so far shows some similarities although the patterns do not fit perfectly. Mutagenicity profiles of TA700x-activity of nitro-aromatic compounds published so far are similar to those of APM collected in Mannheim, Germany, as well as to standard reference materials 1648 and 1649.     

Mutagenic activity of airborne particulate matter as an indicative measure of atmospheric pollution

Mutagenic activity of organic extracts of airborne particulate matter at four different sites within the urban area of the city of Porto Alegre, Brazil, was investigated using the Salmonella/microsome assay, with the Kado microsuspension method. The extracts were obtained by sonication, sequentially extracted according to polarity, with cyclohexane (CX) and dichloromethane (DCM) solvents. The different fractions were tested for mutagenicity with the Salmonella typhimurium strains TA98, TA98NR and TA98/1,8-DNP6, without S9 mix metabolic activation. A positive frameshift mutagenic response was observed for non-polar (CX) and/or moderately polar (DCM) compounds at the different sites. The responses varied at different seasons of the year, and the highest revertants per m3 (rev/m3) values were observed at the site subject to the strongest influence of automotive vehicles (site 3) in spring (17.13 rev/m3) in DCM fractions, and in summer (13.01 rev/m3) in CX fractions. The responses observed for the TA98NR and TA98/1,8-DNP6 strains suggest the contribution of nitrocompounds to the mutagenic activity observed. Although there appears to be an indicative association between the increased mass per unit volume of air (TSP) and the mutagenicity of organic extracts of airborne particulate matter in the present study, the Salmonella/microsome assay was a sensitive method to define areas contaminated by genotoxic compounds, even in samples that present TPS values acceptable by the environmental quality standards established by law.     

Wind-Mediated Spread of Low-Pathogenic Avian Influenza Virus into the Environment during Outbreaks at Commercial Poultry Farms

Avian influenza virus-infected poultry can release a large amount of virus-contaminated droppings that serve as sources of infection for susceptible birds. Much research so far has focused on virus spread within flocks. However, as fecal material or manure is a major constituent of airborne poultry dust, virus-contaminated particulate matter from infected flocks may be dispersed into the environment. We collected samples of suspended particulate matter, or the inhalable dust fraction, inside, upwind and downwind of buildings holding poultry infected with low-pathogenic avian influenza virus, and tested them for the presence of endotoxins and influenza virus to characterize the potential impact of airborne influenza virus transmission during outbreaks at commercial poultry farms. Influenza viruses were detected by RT-PCR in filter-rinse fluids collected up to 60 meters downwind from the barns, but virus isolation did not yield any isolates. Viral loads in the air samples were low and beyond the limit of RT-PCR quantification except for one in-barn measurement showing a virus concentration of 8.48x10⁴ genome copies/m³. Air samples taken outside poultry barns had endotoxin concentrations of ~50 EU/m³ that declined with increasing distance from the barn. Atmospheric dispersion modeling of particulate matter, using location-specific meteorological data for the sampling days, demonstrated a positive correlation between endotoxin measurements and modeled particulate matter concentrations, with an R² varying from 0.59 to 0.88. Our data suggest that areas at high risk for human or animal exposure to airborne influenza viruses can be modeled during an outbreak to allow directed interventions following targeted surveillance.     

Activity and interleukin 1 responsiveness of SV40 enhancer motifs in a rodent immature T cell line.

We have analysed the enhancer activity and the interleukin 1 (IL1) responsiveness of individual motifs of the SV40 enhancer in an immature rodent T cell line, PC60. Transient transfection assays showed that tetramers of GT-I plus GT-IIC motifs, the TC-II or the P motif have significant enhancer activity in PC60, while neither Octamer nor SphI+II motifs have a detectable effect on promoter strength. Two motifs, TC-II and P, strongly respond to stimulation by IL1. DNase I and methylation protection experiments with nuclear extracts show specific footprints in the TC-II region of the SV40 enhancer. Exposure of PC60 cells to IL1 increases their intensity. The TC-II sequence forms several complexes detected in band shift assays. The molecules involved all have similar sequence specificity as NF-kappa B. Surprisingly, band shifts with extracts from control or IL1 treated cells differ only slightly. However, if GTP is added to the binding reactions the intensity of bands formed by extracts from control cells is strongly reduced, whereas extracts from IL1 treated cells form a single retarded complex that co-migrates with NF-kappa B from a pre-B cell line. The results suggest that in PC60 IL1 induces NF-kappa B activity by activating molecules that are already in the nucleus.     

Induction of morphological transformation by coal-dust extract in BALB/3T3 A31-1-13 cell line

The transforming activity of coal dust extracts was studied using BALB/3T3 clone A31-1-13 cells. Coal-dust extracts, both nitrosated and nonnitrosated, induced cell transformation in a dose-response manner. However, the transformation frequency was higher with the nitrosated than with the nonnitrosated extract. All transformed cell lines derived from coal-dust extract-induced foci showed biological characteristics of neoplastic transformation such as loss of contact inhibition and anchorage-independent growth. These results appear to support a hypothesis of coal mine dust causation of gastric cancer in coal miners.