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To explore the impact of different functional groups on Hg(II) adsorption, a range of poly(pyrrole methane)s functionalized by -Cl, -CN, -NH 2 , -OH and -COOH were synthesized and applied to reveal the interaction between different functional groups and mercury ions in water, and the adsorption mechanism was revealed through combined FT-IR, XPS, and DFT calculations. The adsorption performance can be improved to varying degrees by the incorporation of functional groups. Among them, the oxygen-containing functional groups (-OH and -COOH) exhibit stronger affinity for Hg(II) and can increase the adsorption capacity from 180 mg g -1 to more than 1400 mg g -1 at 318 K, with distribution coefficient (K d ) exceeding 10 5  mL g -1 . The variations in the capture and immobilization capabilities of functionalized poly(pyrrole methane)s predominantly stem from the unique interactions between their functional groups and mercury ions. In particular, oxygen-containing -OH and -COOH effectively capture Hg(OH) 2 through hydrogen bonding, and further deprotonate to form the -O-Hg-OH and -COO-Hg-OH complexes which are more stable than those obtained from other functionalized groups. Finally, the ecological safety has been fully demonstrated through bactericidal and bacteriostatic experiments to prove the functionalized poly(pyrrole methane)s can be as an environmentally friendly adsorbent for purifying contaminated water., 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 © 2024 Elsevier Ltd. All rights reserved.)

Nitroalkanes are organic aliphatic hydrocarbon compounds with a nitro moiety that are commonly used as solvents or intermediates to synthesize a variety of organic compounds due to their inherent reactivity. In June 2020, a harmonized classification and labeling (CLH) proposal was submitted to the European Chemicals Agency (ECHA) for the following harmonized carcinogenicity, mutagenicity, and reproductive toxicity ("CMR") classifications for nitromethane (NM), nitroethane (NE), and 1-nitropropane (1-NP): NM Carc. 1B and Repr. 1B; NE Repr. 1B; and 1-NP Repr. 2. In this assessment, a weight of evidence (WoE) evaluation of studies on animal carcinogenicity and reproductive and developmental toxicity, genotoxicity, and mode of action for these three nitroalkanes was performed to critically assess the relevance of the proposed CMR classifications. Overall, the WoE indicates that NM, NE, and 1-NP are not carcinogenic, genotoxic, nor selective reproductive or developmental toxicants. Based on our analysis, classifying NM, NE, and 1-NP as Category 2 reproductive toxicants is most appropriate. Furthermore, not classifying NE and 1-NP with respect to their carcinogenicity is appropriate based on the available studies for this endpoint coupled with negative results in genotoxicity studies, metabolism data, and in silico predictions. We determined that the classification for NM of Carc. 1B is not appropriate, based on the fact that rat mammary and harderian tumors are likely not relevant to humans and lung and liver tumors reported in mice were equivocal in their dose-response and statistical significance., (© 2021 John Wiley & Sons, Ltd.)

Gaseous emission, particulate emission, biological molecules and other harmful substances discharge into the atmosphere from dumpsite environment. The carbon dioxide (CO2) and methane (CH4) content of the gaseous emission from different platforms and offsets of the Awotan dump site were measured. P Sense CO2 Meter AZ 7755 was used to measure the level of carbon dioxide gas, temperature and relative humidity. K60 Gas detector was used to measure the level of methane (CH4) gas and Pm 2.5. Thermo-scientific MIE pDR 1500 PM monitor was used to measure the particulate matter on the dumpsite. The CO2 levels (697±28.84 - 502±2.19) were above the minimum permissible levels of ASHRAE of 400ppm for all platforms at the dumpsite. CH4 levels range (73.33±3.32 - 18.33±4.27) was above the methane explosive limits (MEL) of 15% for all Platforms, however the level at 25m and 50m offsets (14.83±4.11 - 13.83±2.48) was below the MEL for 75m and 100m offsets. PM2.5 levels were lower in the morning and peaked in the afternoon at Platform 5, 6 and 9 locations with values of 62.76±6.03, 63.9±11.37 and 32.06±3.89 respectively which is not within the WHO minimum permissible limit of 25µg/m3. There was a significant positive correlation between CO2 and CH4 (r=0.7558, p=0.028) but no significant correlation between CO2 and other meteorological parameters (temperature and humidity) (r=-0.1309, p=0.67and r=0.09644, p=0.754). The carbon dioxide and methane content of the gaseous emission from the Awotan dump site are potential health hazard, hence the need for an engineering design that will reduce the quantum of the emission thereby reducing the hazard.

With the progress of biotechnological research and improvements made in bioprocessing with pure cultures, microbial consortia have gained recognition for accomplishing biological processes with improved effectiveness. Microbes are indispensable tool in developing bioprocesses for the production of bioenergy and biochemicals while utilizing renewable resources due to technical, economic and environmental advantages. They communicate with specific cohorts in close proximity to promote metabolic cooperation. Use of positive microbial associations has been recognized widely, especially in food industries and bioremediation of toxic compounds and waste materials. Role of microbial associations in developing sustainable energy sources and substitutes for conventional fuels is highly promising with many commercial prospects. Detoxification of chemical contaminants sourced from domestic, agricultural and industrial wastes has also been achieved through microbial catalysis in pure and co-culture systems. Methanotrophs, the sole biological sink of greenhouse gas methane, catalyze the methane monooxygenasemediated oxidation of methane to methanol, a high energy density liquid and key platform chemical to produce commodity chemical compounds and their derivatives. Constructed microbial consortia have positive effects, such as improved biomass, biocatalytic potential, stability etc. In a methanotroph-heterotroph consortium, non-methanotrophs provide key nutrient factors and alleviate the toxicity from the culture. Non-methanotrophic organisms biologically stimulate the growth and activity of methanotrophs via production of growth stimulators. However, methanotrophs in association with co-cultured microorganisms are in need of further exploration and thorough investigation to study their interaction mode and application with improved effectiveness.

Hydrogen plays a key role in many microbial metabolic pathways in the human gastrointestinal tract (GIT) that have an impact on human nutrition, health and wellbeing. Hydrogen is produced by many members of the GIT microbiota, and may be subsequently utilized by cross-feeding microbes for growth and in the production of larger molecules. Hydrogenotrophic microbes fall into three functional groups: sulfate-reducing bacteria, methanogenic archaea and acetogenic bacteria, which can convert hydrogen into hydrogen sulfide, methane and acetate, respectively. Despite different energy yields per molecule of hydrogen used between the functional groups, all three can coexist in the human GIT. The factors affecting the numerical balance of hydrogenotrophs in the GIT remain unconfirmed. There is increasing evidence linking both hydrogen sulfide and methane to GIT diseases such as irritable bowel syndrome, and strategies for the mitigation of such health problems through targeting of hydrogenotrophs constitute an important field for further investigation.

Background: Schisandrin B (Sch B) a main active component of Schisandra chinensis , has been shown to act as a liver protectant via activation of the Nrf2 pathway. Nevertheless, it remains unclear whether its reactive metabolite is responsible for Nrf2 activation; also, the effects of its reactive metabolite on liver function are still unknown., Methods: The present study determined and identifed the carbene reactive metabolite of Sch B in human and mice liver microsomes. Its roles in activating Nrf2 pathway and modifying macromolecules were further explored in human liver microsomes. Moreover the potential cytotoxicity and hepatoxicity of carbene on HepG-2 and mice were also investigated., Results: In the present study, cytochromes P450 (CYP450s) metabolized Sch B to carbene reactive metabolite, which, with the potential to modify peptides, were identifed and observed in human and mice liver microsomes. Moreover, the relevance of carbene in Nrf2 activation was verifed by co-incubation in the presence of CYP450 inhibitors in HepG-2 cells, as well as by molecular docking study of carbene and Keap1. Additionally, the cytotoxicity of Sch B on HepG-2 cells was signifcantly aggravated by CYP450 inducer (with LD 50 decreasing from 63 to 21 µM) and signifcantly alleviated by CYP450 inhibitor and glutathione (with LD 50 increasing from 63 µM to 200 µM). Besides, after oral administration of mice with Sch B (25-100 mg/kg) for 21 days, only the highest dose induced mild hepatotoxicity, which was accompanied by increasing the aminotransferase activity and centrilobular hepatocellular infltration of lymphocytes. In addition, upregulation of CYP450 activity; Nrf2, NQO-1, and GST expression; and glutathione level was observed in Sch B treatment groups., Conclusion: The present study revealed that CYP450s mediate the conversion of Sch B to carbene, which subsequently binds to Keap1 and elicits Nrf2 pathway, which could further increase the elimination of carbene and thus exhibit a less harmful effect on mice liver., Competing Interests: Disclosure The authors report no conflicts of interest in this work.

CO 2 boilers/direct heating systems used in greenhouses often lead to incomplete combustion, which results in the formation of hazardous gases, such as carbon monoxide (CO), nitroxide (NO X ) and other hydrocarbons. In this study, strawberry plants that were grown on rockwool cubes were transferred to airtight bottles and treated with CO, NO X , CH 4 and C 3 H 8 gases for 1-48 hours. Oxidative damage due to hazardous gases was observed, as indicated by H 2 O 2 and [Formula: see text] determination. Photosynthetic pigments were reduced, and stomatal guard cells were damaged and remained closed compared to the control. The activity of other photosynthetic parameters was negatively related to hazardous gases. Reduction in the expression of multiprotein complexes was highly observed under hazardous gas treatments. This study highlighted that hazardous gases (CO, NO X , CH 4 and C 3 H 8 ) emitted due to incomplete combustion of CO 2 fertilization units/or direct heating systems resulted in the formation of ROS in shoots and limited photosynthetic metabolism. We predicted that major steps must be incorporated to improve the efficiency of CO 2 boiler/heating systems to decrease the emission of these hazardous gases and other hydrocarbons and to reduce the observed risks that are key to the reduction of crops.

Halonitromethanes (HNMs) as one typical class of nitrogenous disinfection byproducts have been widely found in drinking water. In vitro test found HNMs could induce higher cytotoxicity and genotoxicity than trihalomethanes and haloacetic acids. However, data on toxic effect from in vivo experiment is limited. In this study, bromonitromethane (BNM), bromochloronitromethane (BCNM) and trichloronitromethane (TCNM) were chosen as target HNMs, and exposed to mice for 30 d. Hepatic toxicity and serum metabolic profiles were determined to reveal toxic effects and mechanisms of the three HNMs. Results showed the three HNMs significantly decreased relative liver weight, indicating liver is one of the target organs. Further, the three HNMs exposure damaged hepatic antioxidant defense system, and increased oxidative DNA damage. Nuclear magnetic resonance based metabolomics analysis found amino acid metabolism and carbohydrate metabolism were disturbed by HNMs exposure. Some metabolites in these metabolisms are related to oxidative stress and damage. Combined with above results, BNM had the highest toxicity, followed by BCNM and TCNM, indicating bromo-HNMs had higher toxicity than chloro-HNMs. Induction of oxidative stress is one of the toxicity mechanisms of HNMs. This study firstly provides the insight into in vivo toxicity of HNMs and their underlying mechanisms based on metabolomics methods, which is very useful for their health risk assessment in drinking water., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A series of organometallic Au I N-heterocyclic carbene (NHC) complexes was synthesized and characterized for anticancer activity in four human cancer cell lines. The compounds' toxicity in healthy tissue was determined using precision-cut kidney slices (PCKS) as a tool to determine the potential selectivity of the gold complexes ex vivo. All evaluated compounds presented cytotoxic activity toward the cancer cells in the nano- or low micromolar range. The mixed Au I NHC complex, (tert-butylethynyl)-1,3-bis-(2,6-diisopropylphenyl)imidazol-2-ylidene gold(I), bearing an alkynyl moiety as ancillary ligand, showed high cytotoxicity in cancer cells in vitro, while being barely toxic in healthy rat kidney tissues. The obtained results open new perspectives toward the design of mixed NHC-alkynyl gold complexes for cancer therapy., (© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Methane has been associated with constipation-predominant irritable bowel syndrome, slowing intestinal transit time by augmenting contractile activity. However, the precise mechanism underlying this effect remains unclear. Therefore, we investigated the mechanisms underlying the effect of methane on contractile activity, and whether such effects are mediated by nerve impulses or muscular contraction., Methods: We connected guinea pig ileal muscle strips to a force/tension transducer and measured amplitudes of contraction in response to electrical field stimulation (EFS; 1, 2, 8, 16 Hz) following methane infusion in the presence of tetradotoxin (TTX), atropine, guanethidine, or GR 113808. We then performed calcium imaging using Oregon Green 488 BAPTA-1 AM in order to visualize changes in calcium fluorescence in response to EFS following methane infusion in the presence of TTX, atropine, or a high K + solution., Key Results: Methane significantly increased amplitudes of contraction (P<.05), while treatment with TTX abolished such contraction. Methane-induced increases in amplitude were inhibited when lower-frequency (1, 2 Hz) EFS was applied following atropine infusion (P<.05). Neither guanethidine nor GR 113808 significantly altered contraction amplitudes. Methane significantly increased calcium fluorescence, while this increase was attenuated following atropine infusion (P<.05). Although calcium fluorescence was increased by the high K + solution under pretreatment with TTX, the intensity of fluorescence remained unchanged after methane infusion., Conclusions and Inferences: The actions of methane on the intestine are influenced by the cholinergic pathway of the enteric nervous system. Our findings support the classification of methane as a gasotransmitter., (© 2017 John Wiley & Sons Ltd.)

Two gold(I) N-heterocyclic carbene complexes 1a and 1b were tested for their anti-trypanosomal activity against Trypanosoma brucei parasites. Both gold compounds exhibited excellent anti-trypanosomal activity (IC 50 =0.9-3.0nM). The effects of the gold complexes 1a and 1b on the T. b. brucei cytoskeleton were evaluated. Rapid detachment of the flagellum from the cell body occurred after treatment with the gold complexes. In addition, a quick and complete degeneration of the parasitic cytoskeleton was induced by the gold complexes, only the microtubules of the detached flagellum remained intact. Both gold compounds 1a and 1b feature selective anti-trypanosomal agents and were distinctly more active against T. b. brucei cells than against human HeLa cells. Thus, the gold complexes 1a and 1b feature promising drug candidates for the treatment of trypanosome infections such as sleeping sickness (human African Trypanosomiasis caused by Trypanosoma brucei parasites)., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Emissions from solid fuels used for cooking cause ~4 million premature deaths per year. Advanced solid-fuel cookstoves are a potential solution, but they should be assessed by appropriate performance indicators, including biological effects., Objective: We evaluated two categories of solid-fuel cookstoves for eight pollutant and four mutagenicity emission factors, correlated the mutagenicity emission factors, and compared them to those of other combustion emissions., Methods: We burned red oak in a 3-stone fire (TSF), a natural-draft stove (NDS), and a forced-draft stove (FDS), and we combusted propane as a liquified petroleum gas control fuel. We determined emission factors based on useful energy (megajoules delivered, MJd) for carbon monoxide, nitrogen oxides (NOx), black carbon, methane, total hydrocarbons, 32 polycyclic aromatic hydrocarbons, PM2.5, levoglucosan (a wood-smoke marker), and mutagenicity in Salmonella., Results: With the exception of NOx, the emission factors per MJd were highly correlated (r ≥ 0.97); the correlation for NOx with the other emission factors was 0.58-0.76. Excluding NOx, the NDS and FDS reduced the emission factors an average of 68 and 92%, respectively, relative to the TSF. Nevertheless, the mutagenicity emission factor based on fuel energy used (MJthermal) for the most efficient stove (FDS) was between those of a large diesel bus engine and a small diesel generator., Conclusions: Both mutagenicity and pollutant emission factors may be informative for characterizing cookstove performance. However, mutagenicity emission factors may be especially useful for characterizing potential health effects and should be evaluated in relation to health outcomes in future research. An FDS operated as intended by the manufacturer is safer than a TSF, but without adequate ventilation, it will still result in poor indoor air quality., Citation: Mutlu E, Warren SH, Ebersviller SM, Kooter IM, Schmid JE, Dye JA, Linak WP, Gilmour MI, Jetter JJ, Higuchi M, DeMarini DM. 2016. Mutagenicity and pollutant emission factors of solid-fuel cookstoves: comparison with other combustion sources. Environ Health Perspect 124:974-982; http://dx.doi.org/10.1289/ehp.1509852.

A room-temperature continuous-wave (CW) quantum cascade laser (QCL)-based methane (CH4) sensor operating in the mid-infrared near 8 μm was developed for continuous measurement of CH4 concentrations in ambient air. The well-isolated absorption line (7F2,4 ← 8F1,2) of the ν4 fundamental band of CH4 located at 1255.0004 cm(-1) was used for optical measurement of CH4 concentration by direct absorption in a White-type multipass cell with an effective path-length of 175 m. A 1σ (SNR = 1) detection limit of 33.3 ppb in 218 s was achieved with a measurement precision of 1.13%. The developed sensor was deployed in a campaign of measurements of time series CH4 concentration on a site near a suburban traffic road in Dunkirk (France) from 9th to 22nd January 2013. An episode of high CH4 concentration of up to ~3 ppm has been observed and analyzed with the help of meteorological parameters combined with back trajectory calculation using the Hybrid Single Particle Lagrangian Integrated Trajectory (HYSPLIT) model of NOAA.

Chloropicrin (trichloronitromethane) is a widely used soil fumigant and an old chemical warfare agent. The metabolism of chloropicrin is not well known in mammals but nitromethane has been shown to be one of its main metabolites. Here, a fast and simple headspace gas chromatography with mass spectrometry method was applied for the measurement of nitromethane from aqueous samples. The analytical method was validated using stable isotope labeled internal standard and a small sample volume of 260 μL. No conventional sample preparation steps were needed. The method was accurate (relative standard deviations ≤1.5%) and linear (R(2) = 0.9996) within the concentration range of 0.1-6.0 μg/mL. This method was used to measure nitromethane in in vitro incubations with human and pig liver cell fractions containing enzymes for xenobiotic metabolism, exposed to chloropicrin. The results indicate that the presence of glutathione is necessary for the formation of nitromethane from chloropicrin. Also, nitromethane was formed mostly in liver cytosol fractions, but not in microsomal fractions after the incubation with chloropicrin. Our results suggest that although nitromethane is not the unequivocal biomarker of chloropicrin exposure, this method could be applied for screening the elevated levels in humans after chloropicrin exposure., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

In this study, a series of new 4,4'-diaminotriphenylmethanes was efficiently synthesized from aromatic aldehydes and 2,5-dimethoxybenzenamine under microwave irradiation in the presence of Sc(OTf)3 as a catalyst. Antiproliferative activity was assessed by using the MCF-7 estrogen receptor (ER)-positive breast cancer cell line, and antagonist/agonist transcriptional activities were determined. Docking studies and competition studies of triphenylmethanes and radiolabeled estradiol determined that these compounds do not bind the ER, indicating that triphenylmethane-induced changes in proliferative and transcriptional activities differ from conventional mechanisms of action triggered by other selective ER modulators., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Triarylmethanes (TRAMs) and thiophene containing trisubstituted methanes (TRSMs) have been reported by us, having potential against Mycobacterium tuberculosis and Mycobacterium fortuitum strains, respectively. Further, extension through synthesis and biological evaluation of novel TRSMs resulted into an identified lead 36 (S006-830) [(diisopropyl-(2-{4-[(4-methoxy-phenyl)- thiophen-2-yl-methyl]-phenoxy}-ethyl)-amine)] with MIC: 1.33 mg/L, non-toxic against Vero C-1008 cell line with selectivity index >10, ex vivo efficacy equivalent to first line TB drugs-isoniazid (INH), rifampicin (RFM) and pyrazinamide (PZA) in the mouse and human macrophages, and lung CFU count of 2.2 × 10(7) (approximately 15 fold lesser than untreated mice, 31 × 10(7)) with efficacies comparable to ethambutol (EMB) (1.27 × 10(7)) and PZA (1.9 × 10(7)). Further, S006-830 also showed potent bactericidal activity against multi-drug resistant and single-drug resistant clinical isolates of M. tuberculosis., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

Recent research on the agricultural impacts of climate change has primarily focused on the roles of temperature and precipitation. These studies show that India has already been negatively affected by recent climate trends. However, anthropogenic climate changes are a result of both global emissions of long-lived greenhouse gases (LLGHGs) and other short-lived climate pollutants (SLCPs). Two potent SLCPs, tropospheric ozone and black carbon, have direct effects on crop yields beyond their indirect effects through climate; emissions of black carbon and ozone precursors have risen dramatically in India over the past three decades. Here, to our knowledge for the first time, we present results of the combined effects of climate change and the direct effects of SLCPs on wheat and rice yields in India from 1980 to 2010. Our statistical model suggests that, averaged over India, yields in 2010 were up to 36% lower for wheat than they otherwise would have been, absent climate and pollutant emissions trends, with some densely populated states experiencing 50% relative yield losses. [Our point estimates for rice (-20%) are similarly large, but not statistically significant.] Upper-bound estimates suggest that an overwhelming fraction (90%) of these losses is due to the direct effects of SLCPs. Gains from addressing regional air pollution could thus counter expected future yield losses resulting from direct climate change effects of LLGHGs.

In mammals, it has been shown that halomethanes (HM) are bioactivated by enzymes such as CYP 2E1 and the theta isoform of GST to produce reactive metabolites. However, in fish, little information is available, although HM can form autochthonously in aquatic environments. This study assessed the effect of HM in dusky splitfin (Goodea gracilis) from three lakes of the Valley of Mexico by analysing specific HM biomarkers as well as a broad range of biomarkers. The concentration of HM was a function of its half-life (higher in deep waters), while its precursors and solar radiation are secondary factors that determine its concentration. The kidney showed higher basal metabolism than the liver, probably because of its function as a haematopoietic and filtration organ. Using integrated biological response version 2 (IBRv2), it was found that the hepatic and renal O₂· content is a pro-oxidant force capable of inducing oxidative stress (ROOH, TBARS and RC=O). Early damage was found to be dependent on low concentrations of HM in Major Lake, whereas late damage was observed in fish exposed to higher concentrations of HM in Zumpango Lake and Ancient Lake. The activities of enzymes involved in antioxidant defence seemed to be inefficient. The quantitative assessment of biomarkers (ANOVA) and the estimate of parameter A obtained from IBRv2 provided different information. However, the data support the greater predictive power of IBRv2, but it requires a series of interrelated biomarkers to infer these possibilities. G. gracilis presents marked patterns of adaptation, which are dependant on the HM concentrations in environmental mixtures, although the response is complex and many toxicants could induce similar responses.

Halomethanes (HMs) can be formed during the chlorination process to obtain drinking water. In liver cells, HMs had been shown to be mutagenic and carcinogenic; however, their bioactivation by CYP 2E1 and GSTT1 is required. Although inhalation is the most common pathway of exposure, reports on the toxic effects induced by HMs in human lung are contradictory. The aim of this study was therefore to evaluate in vitro cytotoxicity and cell proliferation induced by CH(2)Cl(2), CHCl(3) and BrCHCl(2) in human lung NL20-TA epithelial cells and MRC-5 fibroblasts, and their relationship with CYP 2E1 and GSTT1 activity. High concentrations of these HMs induced cytotoxicity, particularly in cells treated with BrCHCl(2). Low concentrations of BrCHCl(2) stimulated hyperproliferation of fibroblasts, the most probable consequence of which is regenerative proliferation related to collagen induction. Fibroblasts exposed to BrCHCl(2) exhibited low levels of CYP 2E1 activity suggesting that released bromine is able to alter this activity by affecting the active site or auto regulating the activity itself. GSTT1 was up to ten times more active than CYP 2E1 in both cell lines, indicating that potential lung damage is due to formation of pro-carcinogens such as formaldehyde.

Objectives: Mycobacterium fortuitum causes opportunist non-tubercular infection in humans. Chronic infection of M. fortuitum has been clinically documented and requires prolonged chemotherapy. The objectives of this study were to characterize acute and persistent infection of M. fortuitum in a murine infection model and to screen thiophene-containing trisubstituted methanes active against both acute and persistent infection., Methods: A murine infection model of M. fortuitum was used. Bacillary count, bioluminescence, disease symptoms, host immune response, drug susceptibility and mortality were measured. Reactivation of persistent bacilli was induced by dexamethasone. Trisubstituted methanes containing thiophene rings were synthesized and screened in vitro by agar dilution and BACTEC assay and in mice. Cytotoxicity was tested with Vero monkey kidney cells using a resazurin assay., Results: The acute infection in mice was marked by a 3 log rise in viable counts, the appearance of disease symptoms and a rise in the Th1 immune response. Bacilli were susceptible to fluoroquinolones. This was followed by persistent infection, in which disappearance of disease symptoms, a decline in Th1 response and non-susceptibility to fluoroquinolones was observed. When the mice were immunocompromised on day 40 post-infection (persistent state) by dexamethasone, a rise in viable counts, symptoms and susceptibility to fluoroquinolones and a prominent Th1 response reappeared. Two lead compounds were found that cleared the mice of bacilli in acute infection and caused a 2.29-2.99 log reduction in cfu of persistent bacilli., Conclusions: The study established acute and persistent infection in mice and identified two promising anti-M. fortuitum compounds with a selectivity index >10.

Halomethanes (HMs) are produced autochthonously in water bodies through the action of ultraviolet light in the presence of HM precursors, such as dissolved organic carbon and halogens. In mammals, toxic effects induced by HMs are diverse and include oxidative stress, which is also induced by divalent and polyvalent metals; however, in fish little information is available on HM metabolism and its possible consequences at the population level. In the present study, high CYP 2E1 and GST theta-like activities were found in viscera of the Toluca silverside Chirostoma riojai from Lake Zumpango (LZ; central Mexico). Formaldehyde, one of the HM metabolites, was correlated with CYP 2E1 activity and also induced lipid peroxidation in viscera. Hepatic CYP 2E1 activity was correlated with GST theta-like activity, suggesting the coupling of both pathways of HM bioactivation and its consequent oxidative damage. Sediment metals, among others, were also responsible for oxidative stress, particularly iron, lead, arsenic and manganese. However, under normal environmental conditions, the antioxidant enzymes of this species sustain catalysis adapted to oxidative stress. Findings suggest that this fish species apparently has mechanisms of adaptation and recovery that enable it to confront toxic agents of natural origin, such as metals and other substances formed through natural processes, e.g., HMs. This has allowed C. riojai to colonize LZ despite the high sensitivity of this species to xenobiotics of anthropogenic origin.

In the present study, we aimed to determine the oxidative damage in rat heart tissue induced by ruthenium(II)-NHC (Ru) and gold(I)-NHC (Au) complexes which have anticarcinogenic effects and not used clinically yet. For this purpose, 35 Sprague-Dawley rats were randomly divided into 5 equal groups. In the control group, rats treated with saline, Ru and Au complexes were intraperitoneally given high (10 mg/kg) and low (5 mg/kg) doses as only one administration. The animals were killed, and heart tissues were taken on day 10 of the drug administration for the determination of the biochemical parameters (malondialdehyde, superoxide dismutase, reduced glutathione and catalase levels). It was determined that both Ru and Au complexes treatment significantly caused oxidative damage compared to the control group. Additionally, it was shown that Au treatment caused more adverse effects than Ru treatment. Also, it was clearly found that the occurred effects were generally determined in a dose-dependent manner. In conclusion, when these compounds synthesized for the treatment of cancer were used, they caused oxidative damage in heart tissue. However, Ru complex could be preferred for cancer treatment in terms of user safety.

Cis-platin and other platinum complexes are important chemotherapeutic agents useful in the treatment of several cancers. However, therapeutic usage of cis-platin and other platinum complex are limited by their undesirable side effects including cardiotoxicity. In this context, we aimed to compare the damage caused in heart by cis-platin and novel platinum-N-heterocyclic carbene (Pt-NHC) complex. For this purpose, 35 Sprague-Dawley rats were divided randomly into five equal groups (n = 7 for each group). Cis-platin and novel Pt-NHC complex were intraperitoneally administered at a single dose of 5 mg/kg and 10 mg/kg and then sacrificed 10 days after this treatment. The heart tissues were taken from all rats for determination of oxidative and myocardial damage. Cis-platin and novel Pt-NHC complex caused oxidative and histological damage in the heart tissue in a dose-dependent manner (p < 0.05). On the other hand, at the same dose levels, cis-platin caused lower oxidative and histological damage in heart tissue compared to novel Pt-NHC complex. These results suggest that novel Pt-NHC complex is more cardiotoxic than cis-platin.

Of the many sources of urban greenhouse gas (GHG) emissions, solid waste is the only one for which management decisions are undertaken primarily by municipal governments themselves and is hence often the largest component of cities' corporate inventories. It is essential that decision-makers select an appropriate quantification methodology and have an appreciation of methodological strengths and shortcomings. This work compares four different waste emissions quantification methods, including Intergovernmental Panel on Climate Change (IPCC) 1996 guidelines, IPCC 2006 guidelines, U.S. Environmental Protection Agency (EPA) Waste Reduction Model (WARM), and the Federation of Canadian Municipalities-Partners for Climate Protection (FCM-PCP) quantification tool. Waste disposal data for the greater Toronto area (GTA) in 2005 are used for all methodologies; treatment options (including landfill, incineration, compost, and anaerobic digestion) are examined where available in methodologies. Landfill was shown to be the greatest source of GHG emissions, contributing more than three-quarters of total emissions associated with waste management. Results from the different landfill gas (LFG) quantification approaches ranged from an emissions source of 557 kt carbon dioxide equivalents (CO2e) (FCM-PCP) to a carbon sink of -53 kt CO2e (EPA WARM). Similar values were obtained between IPCC approaches. The IPCC 2006 method was found to be more appropriate for inventorying applications because it uses a waste-in-place (WIP) approach, rather than a methane commitment (MC) approach, despite perceived onerous data requirements for WIP. MC approaches were found to be useful from a planning standpoint; however, uncertainty associated with their projections of future parameter values limits their applicability for GHG inventorying. MC and WIP methods provided similar results in this case study; however, this is case specific because of similarity in assumptions of present and future landfill parameters and quantities of annual waste deposited in recent years being relatively consistent.

The Borgwaldt RM20S(®) smoking machine enables the generation, dilution, and transfer of fresh cigarette smoke to cell exposure chambers, for in vitro analyses. We present a study confirming the precision (repeatability r, reproducibility R) and accuracy of smoke dose generated by the Borgwaldt RM20S(®) system and delivery to exposure chambers. Due to the aerosol nature of cigarette smoke, the repeatability of the dilution of the vapor phase in air was assessed by quantifying two reference standard gases: methane (CH(4), r between 29.0 and 37.0 and RSD between 2.2% and 4.5%) and carbon monoxide (CO, r between 166.8 and 235.8 and RSD between 0.7% and 3.7%). The accuracy of dilution (percent error) for CH(4) and CO was between 6.4% and 19.5% and between 5.8% and 6.4%, respectively, over a 10-1000-fold dilution range. To corroborate our findings, a small inter-laboratory study was carried out for CH(4) measurements. The combined dilution repeatability had an r between 21.3 and 46.4, R between 52.9 and 88.4, RSD between 6.3% and 17.3%, and error between 4.3% and 13.1%. Based on the particulate component of cigarette smoke (3R4F), the repeatability (RSD = 12%) of the undiluted smoke generated by the Borgwaldt RM20S(®) was assessed by quantifying solanesol using high-performance liquid chromatography with ultraviolet detection (HPLC/UV). Finally, the repeatability (r between 0.98 and 4.53 and RSD between 8.8% and 12%) of the dilution of generated smoke particulate phase was assessed by quantifying solanesol following various dilutions of cigarette smoke. The findings in this study suggest the Borgwaldt RM20S(®) smoking machine is a reliable tool to generate and deliver repeatable and reproducible doses of whole smoke to in vitro cultures.

N-Heterocyclic carbene (NHC) ligands have attracted great interest over the last decade for their use in the design of homogenous catalysts. NHC-based metal complexes have interesting potential biomedical applications, such as in antimicrobial and cancer therapy, which are beginning to be explored more fully. We have studied here the oxidant activities of a series of Ru(II) complexes in vitro and zebrafish (Danio rerio) have been used as a model in vivo to investigate and characterize the toxicity of some of these compounds. Dual behavior was observed for the NHC-based complexes as they behaved as antioxidants at low concentrations but showed pro-oxidant capacity at higher concentrations. Zebrafish embryos were exposed to Ru(II) complexes under several different conditions (0 or 24 h postfertilization, with or without the chorion) and various parameters, such as viability, edema, heart rate, blood coagulation, pigmentation, scoliosis, malformation, and hatching, were tested. In general, zebrafish embryos were not harmed by exposure to Ru(II) complexes whatever the experimental conditions. Several toxicity profiles were observed depending upon the chemical structure of the compound in question. Their characteristics as pro-oxidant and/or antioxidant agents together with their biosafety may point to their having biomedical applications as antitumoral or neuroprotective drugs.

Despite the all precautions, underground coal mining is one of the dangerous industries owing to fatal occupational accidents. Accidents are complicated events to which many factors effect on their formation and preventing them is only possible by the analyses of the accident occurred in past and by straight evaluation of the obtained results. In this study, hierarchical loglinear analysis method was implemented to occupational fatalities occurred in the period of 1980-2004 in the five underground coal mines of Turkish Hardcoal Enterprises which has the most important coal production areas in Turkey. The accident records were evaluated and the main factors affecting the accidents were defined as mine, miners' age, occupation, and accident type. By taking into account the sub factors of the main factors, multi way contingency tables were prepared and thus, the probabilities might effect fatality accidents were investigated. At the end of this study, it was found that the mostly affected job group by the fatality accidents was the production workers and additionally, these workers were mostly exposed to roof collapses and methane explosions. Moreover, important accident risk factors and the occupational job groups which have high probability to be exposed to these risk factors were determined and important information about decreasing the accidents in the underground coal mines were presented.

During carcinogenesis, methylation of the C-5 position of cytosines in the promoter region of tumor suppressor genes is often observed. Enzymatic DNA methylation is a widely accepted mechanism for this phenomenon. It is interesting to propose a free radical mechanism for 5-methyldeoxycytidine (m(5)dC) production, because the C-5 position of cytosine is an active site for free radical reactions. When deoxycytidine (dC) and cumene hydroperoxide (CuOOH), a tumor promoter and a methyl radical producer, were reacted in the presence of ferrous ion at pH 7.4, the formation of m(5)dC was observed. The same reaction also proceeded with t-butyl hydroperoxide (BuOOH). The formation of m(5)dC was also observed in DNA by the CuOOH treatment. This is the first report of chemical DNA methylation at cytosine C-5 by environmental tumor promoters. We propose here that this reaction is one of the important mechanisms of de novo DNA methylation during carcinogenesis, because methyl radicals are produced by the biotransformation of various endogenous and exogenous compounds.

The cytotoxicity and genotoxicity of nitromethane and its halogenated analogues in mammals raise concerns about potential toxicity to humans. This study shows that halonitromethanes are not stable in human blood and undergo dehalogenation to form nitromethane. We quantified nitromethane in human blood using solid-phase microextraction (SPME) headspace sampling coupled with gas chromatography (GC) and high resolution mass spectrometry (HRMS). The limit of detection was 0.01 microg/L with a linear calibration curve spanning 3 orders of magnitude. This method employs isotope dilution to precisely quantify trace amounts of nitromethane (coefficient of variation <6%). At three spiked concentrations of nitromethane, method accuracy ranged from 88 to 99%. We applied this method to blood samples collected from 632 people with no known occupational exposure to nitromethane or halonitromethanes. Nitromethane was detected in all blood samples tested (range: 0.28-3.79 microg/L, median: 0.66 microg/L). Time-course experiments with trichloronitromethane- and tribromonitromethane-spiked blood showed that nitromethane was the major product formed (1 nmole tribromonitromethane formed 0.59 nmole of nitromethane, whereas 1 nmole trichloronitromethane formed 0.77 nmole nitromethane). Nitromethane may form endogenously from peroxynitrite: nitromethane concentrations increased proportionately in blood samples spiked with peroxynitrite. Blood nitromethane can be a biomarker of exposure to both nitromethane and halonitromethanes. This sensitive, accurate, and precise analytical method can be used to determine baseline blood nitromethane level in the general population. It can also be used to study the health impact from exposure to nitromethane and halonitromethanes in occupational environments and to assess trichloronitromethane (chloropicrin) exposure in chemical terrorism investigations.

Centralised treatment of domestic sewage coming from A Coruña city (250000 inhabitants) and several medium and small size villages (up to 15000 inhabitants) together with industrial estate effluents was planned. The area population is about 380000 inhabitants and the overall effluent will reach about 600000 equivalent inhabitants. The characteristics of different streams will determine the treatment alternatives and pre-treatment requirements for individual streams. Domestic sewage from A Coruña showed medium to high organic content (741 mg l(-1) of COD), while domestic sewage from small villages (Lorbé, Oleiros) and industrial estates (Sabón, A Grela) showed a high variability in organic content, with COD concentration ranging from 100 to 4000 mg L(-1) or more. About 50-65% COD corresponded to suspended solids, although some industrial effluents showed lower values. Characterized industrial effluents had lower concentration of nutrients (nitrogen and phosphorus) and higher concentration of sulphate and alkalinity than domestic sewage. Anaerobic biodegradability was about 80% for domestic sewage, and ranged from 20 to 100% for industrial estate effluents. Furthermore, some industrial streams showed high microbial toxicity. The results indicate that anaerobic treatment of these effluents is of interest. However, specific pre-treatment for some individual streams or the adoption of minimisation and prevention measures in order to reach total or partial detoxification is suggested.

Emissions of methane (CH4) and carbon dioxide (CO2) from spent mycelia of the mold Penicilium notatum and sludge from the effluent treatment facility (ETPS) of a pharmaceutical industry were estimated twice during a two-week composting before vermicomposting. These wastes are dumped in landfills or sometimes used in agricultural fields and no reports are available on their greenhouse gas producing potentials. The solid wastes contained appreciable organic carbon and nitrogen while very high Fe, Mn and Zn were found in ETPS only. Pure wastes did not support germination of Vigna radiata L. while mixing soil with ETPS and spent mycelia at the ratios of 12:1 and 14:1 led to 80% and 50% germination, respectively. The wastes were mixed with cowdung at the ratios of 1:1, 1:3 and 3:1 for composting. Carbon dioxide emissions were always significantly higher than CH4 emissions from all the treatments due to prevalence of aerobic condition during composting. From some treatments, CH4 emissions increased with time, indicating increasing activity of anaerobic bacteria in the waste mixtures. Methane emissions ranged from 21.6 to 231.7 microg m(-2) day(-1) while CO2 emissions were greater than thousand times at 39.8-894.8 mg m(-2) day(-1). The amount of C emitted as CH4-C and CO2-C from ranged from 0.007% to 0.081% of total C composted. Cowdung emitted highest CH4 followed by spent mycelia and ETPS while ETPS emitted more CO2 than spent mycelia but lesser than cowdung. Global warming potential of emitted CH4 was found to be in the range of 10.6-27.7 mg-CO2-equivalent on a 20-year time horizon. The results suggest that pharmaceutical wastes can be an important source of CH4 and CO2 during composting or any other stockpiling under suitable moisture conditions. The waste mixtures were found not suitable for vermicomposting after two weeks composting and earthworms did not survive long in the mixtures.

The NTP chose to initiate studies in fish as an exploration of alternate or additional models for examining chemical toxicity and carcinogenicity. The use of small fish species in carcinogenicity testing offered potential advantages as a bioassay test system, including significant savings in cost and time over rodent studies. Large numbers of small fish could be easily maintained in a limited area. The two species chosen for study were guppy (Poecilia reticulata) and medaka (Oryzias latipes), both of which are hardy, easily maintained, and have a low occurrence of background lesions. The three chemicals chosen for study in fish had already been studied by the NTP in rodents, permitting a comparison of results between the two models. Two of the chemicals used (2,2-bis(bromomethyl)-1,3-propanediol and 1,2,3-trichloropropane) were mutagenic and multisite carcinogens in rats and mice. The third chemical, nitromethane, was nonmutagenic with a more modest carcinogenic response in rodents. Male and female guppies and medaka were exposed to 2,2-bis(bromomethyl)- 1,3-propanediol (greater than 99% pure), nitromethane, (greater than 99% pure), or 1,2,3-trichloropropane (99% pure) in aquaria water for up to 16 months. OVERALL STUDY DESIGN: Groups of approximately 220 guppies (two replicates of 110) were maintained in aquaria water containing nominal concentrations of 0, 24, 60, or 150 mg/L 2,2-bis(bromomethyl)-1,3-propanediol; 0, 10, 30, or 70 mg/L nitromethane; or 0, 4.5, 9.0, or 18.0 mg/L 1,2,3-trichloropropane. Groups of approximately 340 medaka (two replicates of 170) were maintained in aquaria water containing 0, 24, 60, or 150 mg/L 2,2-bis(bromomethyl)-1,3-propanediol; 0, 10, 20, or 40 mg/L nitromethane; or 0, 4.5, 9.0, or 18.0 mg/L 1,2,3-trichloropropane. The overall study durations were 16 months for all guppy studies, 14 months for 2,2-bis(bromomethyl)-1,3-propanediol-exposed medaka, and 13 months for nitromethane- and 1,2,3-trichloropropane-exposed medaka. Ten guppies and 10 medaka from each group replicate were sacrificed at 9 months for histopathologic analysis. Approximately one third of the remaining fish from each group were placed in chemical-free water at 9 months and constituted a stop-exposure study component. The remainder of the fish were exposed for the duration of the study and constituted the core study component. A stop-exposure component was added to determine if stopping the exposure at 9 months and transferring to chemical-free aquaria might allow for better survival and tumor development. The sex of guppies and medaka was determined at histopathologic analysis. 2,2-BIS(BROMOMETHYL)-1,3-PROPANEDIOL - 16-MONTH STUDY IN GUPPIES: 2,2-Bis(bromomethyl)-1,3-propanediol was chronically toxic to guppies in the 60 and 150 mg/L core and stop-exposure groups. Due to mortality, exposure of core study animals in the 150 mg/L group was terminated on day 443, after approximately 64 weeks on study, and fish were maintained in 2,2-bis(bromomethyl)- 1,3-propanediol-free water in the exposure system until the end of the study at 69 weeks. Nominal exposure concentrations of 24, 60, and 150 mg/L provided actual aquaria water exposure concentrations of 20.0, 53.5, and 139.0 mg/L 2,2-bis(bromomethyl)- 1,3-propanediol, respectively. There were no treatment-related differences between the control and exposed groups in body weights or lengths. At 9 months, hepatocellular adenomas occurred in one 24 mg/L male and in one 150 mg/L male. In the core study, the incidence of hepatocellular adenoma or carcinoma (combined) in 150 mg/L males was greater than that in the controls; multiple adenomas occurred in two 150 mg/L males and in one 150 mg/L female. Cholangioma occurred in a small number of exposed males and females. In the stop-exposure study, incidences of hepatocellular adenoma (including multiple) and of hepatocellular carcinoma were greater in 150 mg/L males than in controls. One cholangioma and one cholangiocarcinoma occurred in the 150 mg/L female group. 14-MONTH STUDY IN MEDAKA: Exposure to 2,2-bis(bromomethyl)-1,3-propanediol did not result in any significant reduction in survival, although the mortality of fish was somewhat greater in the 60 and 150 mg/L core study groups than in the control and 24 mg/L groups. After reallocation, mortality of medaka in the 60 and 150 mg/L core groups was slightly increased over the corresponding stop-exposure groups. Nominal exposure concentrations of 24, 60, and 150 mg/L provided actual exposure concentrations of 19.4, 56.9, and 137.8 mg/L 2,2-bis(bromomethyl)- 1,3-propanediol, respectively. Core study animals in the 60 and 150 mg/L groups were significantly larger, in both body length and weight, than control group fish. In the core study, the incidence of hepatocellular adenoma or carcinoma (combined) was increased in 150 mg/L males. Cholangiocarcinomas occurred in a few exposed males and females, with all but one occurring in 150 mg/L fish. One cholangioma occurred in a 150 mg/L female, and one occurred in a control female. In the stop-exposure study, incidences of hepatocellular adenoma or carcinoma (combined) were marginally increased in the 150 mg/L group of males and in the 60 and 150 mg/L groups of females as compared with controls. Cholangiocarcinoma occurred in one male and one female in the 150 mg/L groups and in one control female. NITROMETHANE - 16-MONTH STUDY IN GUPPIES: Although the cause of death could not be confirmed in many cases, mortality in the 70 mg/L groups appeared to indicate that this level of nitromethane exposure was chronically toxic. This is confirmed by the similar survival rate of guppies from all treatments following removal from treatment aquaria and placement in stop-exposure. Due to the high mortality of fish in the 70 mg/L core study groups, these fish were removed from treatment (day 396) and fixed for histological analyses after approximately 57 weeks on study. The controls and other exposed groups were sacrificed at 70 weeks. Nominal exposure concentrations of 10, 30, and 70 mg/L provided actual exposure concentrations of 9.9, 28.7, and 66.4 mg/L nitromethane, respectively. There were no treatment-related differences between the control and exposed groups in body lengths or weights. 13-MONTH STUDY IN MEDAKA: Nitromethane in the aquaria supported a substantial microfaunal growth which, without frequent cleaning, affected water quality and treatment concentrations. To maintain acceptable water quality and treatment concentrations potentially affected by the rapid microfaunal growth, the study aquaria were brushed once and siphoned three times each day. Due to this frequent activity, a number of fish probably died due to mechanical injury. Unfortunately, the cause of death could not be confirmed in many cases; the mortality from this activity is believed to have been approximately uniform among treatments and should not have affected the comparison of survival between treatments. Based on mortality in this study and the previous life-span evaluation, the life phase of this study was terminated approximately 13.5 months after hatching. Nominal exposure concentrations of 10, 20, and 40 mg/L resulted in actual exposure concentrations of 9.3, 20.8, and 41.7 mg/L nitromethane, respectively. No differences between control and exposed groups were found in body lengths or weights at the 9-month interim evaluation. Due to mortality, unequal numbers of fish were distributed among the core study and stop-exposure aquaria at 9 months. Differences in lengths and weights were found at 13 months. The biological significance of this finding is unknown. At 9 months, a single cholangiocarcinoma occurred in a 40 mg/L male. Hepatocellular adenomas occurred in two 20 mg/L males and in one 40 mg/L female. In the core study, one cholangioma occurred in a 20 mg/L male, and cholangiocarcinomas were seen in a few exposed males, but none occurred in control males. 1,2,3-TRICHLOROPROPANE - 16-MONTH STUDY IN GUPPIES: The survival of exposed guppies was less than that of the control group at 9 months. Reduced survival was evident at 6 months in the 18.0 mg/L groups and at 7 months in the 4.5 and 9.0 mg/L groups. Survival was significantly reduced in the 18.0 mg/L core study group within 1 month of the 9-month interim evaluation, and mortality in this group was 42.6% between 9 months and study termination. Nominal exposure concentrations of 4.5, 9.0, and 18.0 mg/L resulted in actual exposure concentrations of 4.4, 8.8, and 18.2 mg/L 1,2,3-trichloropropane, respectively. Guppies in the 18.0 mg/L core study group were significantly longer and weighed more than the controls. Fish in the 18.0 mg/L stop-exposure group also weighed more than the controls. Mortality of fish during the study resulted in unequal numbers of individuals distributed to core study and stop-exposure aquaria at 9 months. This appears to have influenced the length and weight of fish measured at study termination (i.e., the smaller tank population allowed the fish to grow more). Observed differences in weight and length between controls and 18.0 mg/L fish was most likely an artifact of the reduced numbers of fish in the 18.0 mg/L aquaria. At 9 months, multiple hepatocellular adenomas occurred in one 4.5 mg/L male, and one hepatocellular adenoma occurred in a control male. In the core study, increased incidences of cholangiocellular (bile duct) and hepatocellular neoplasms occurred in exposed groups of males and females. Cholangioma and cholangiocarcinoma were seen in several exposed males and females. In the stop-exposure study, increased incidences of hepatocellular neoplasms occurred in 18.0 mg/L males and increased incidences of cholangiocellular (bile duct) neoplasms occurred in 18.0 mg/L females. (ABSTRACT TRUNCATED)

Halonitromethanes (HNMs) are a recently identified class of disinfection by-products (DPBs) in drinking water that are mutagenic in Salmonella and potent inducers of DNA strand breaks in mammalian cells. Here we compared the mutagenic potencies of the HNMs to those of their halomethane (HM) homologues by testing all nine HNMs and seven of the nine HMs (minus bromomethane and chloromethane) under the same conditions (the pre-incubation assay) in Salmonella TA100 +/- S9. We also determined the mutation spectra for several DBPs. In the presence of S9, all nine HNMs, but only three HMs, dibromomethane (DBM), dichloromethane (DCM), and bromochloromethane (BCM), were mutagenic. Only two DBPs of each class were mutagenic in the absence of S9. The HNMs were generally more potent mutagens than their HM homologues, and the brominated forms of both classes of DBPs were more mutagenic and cytotoxic than their chlorinated homologues. The HNMs were at least 10 times more cytotoxic than the HMs, and the cytotoxicity rankings in the presence of S9 were similar for the HNMs and the HMs. The addition of a nitro-group to BCM did not change the mutation spectra significantly, with both homologues inducing primarily (55-58%) GC --> AT transitions. The greater cytotoxic and mutagenic activities of the HNMs relative to the HMs are likely due to the greater intrinsic reactivity conferred by the nitro-group. Energy calculations predicted increased reactivity with increasing bromination and greater reactivity of the HNMs versus the HMs (Elumo values were approximately 20 kcal/mol lower for the HNMs compared to their HM homologues). Given that the HNMs also are potent genotoxins in mammalian cells [Environ. Sci. Technol. 38 (2004) 62] and are more mutagenic and 10x more cytotoxic in Salmonella than the HMs, whose levels are regulated in drinking water, further study of their occurrence and potential health effects is warranted.

Background: Our past studies have shown that cigarette smoke inhibits oviductal functioning in vivo and in vitro. The goals in this study were to identify pyrazine derivatives in cigarette smoke solutions that inhibit ciliary beat frequency, oocyte pickup rate, and infundibular smooth muscle contraction in the hamster oviduct and to determine their lowest observable adverse effect levels (LOAELs) using in vitro bioassays., Methods: MS smoke solutions were fractionated using solid phase extraction cartridges and the fractions were both tested on the hamster oviduct in vitro and analyzed by gas chromatography-mass spectrometry to identify individual pyrazine derivatives. Commercial pyrazine standards were purchased, assayed for purity, and tested in dose-response studies on hamster oviducts. The LOAEL and efficacy were determined for each compound in the in vitro bioassays. Statistical significance was determined using the Student's t-Test where p < 0.05., Results: The LOAELs for the most inhibitory pyrazine derivatives in the ciliary beat frequency, oocyte pickup rate, and infundibular smooth muscle contraction assays were as follows: for pyrazine (1 picomolar, 10 picomolar, and 1 nanomolar); for 2-methylpyrazine (1 picomolar, 10 picomolar, and 10 picomolar); and for 2-ethylpyrazine (1 picomolar, 10 picomolar, and 1 picomolar). Six of the seven pyrazine derivatives tested (pyrazine, 2-methylpyrazine, 2-ethylpyrazine, 2-methoxy-3-methylpyrazine, 2,5-dimethylpyrazine, and 2,3,5-trimethylpyrazine) were inhibitory in picomolar or nanomolar doses in all three bioassays, while the seventh derivative, 2,6-dimethylpyrazine, had LOAELs in the nanomolar to micromolar range., Conclusion: This work shows that very low doses of pyrazines significantly inhibit proper oviductal functioning, raising questions regarding the safety of these compounds in cigarettes and other consumer products.

Halonitromethanes are drinking water disinfection byproducts that have recently received a high priority for health effects research from the U.S. Environmental Protection Agency (EPA). Our purpose was to identify and synthesize where necessary the mixed halonitromethanes and to determine the chronic cytotoxicity and the acute genotoxicity of these agents in mammalian cells. The halonitromethanes included bromonitromethane (BNM), dibromonitromethane (DBNM), tribromonitromethane (TBNM), bromochloronitromethane (BCNM), dibromochloronitromethane (DBCNM), bromodichloronitromethane (BDCNM), chloronitromethane (CNM), dichloronitromethane (DCNM), and trichloronitromethane (TCNM). Low- and high-resolution gas chromatography/mass spectrometry (GC/MS) was used to identify the mixed chloro-bromonitromethanes in finished drinking waters, and analytical standards that were not commercially available were synthesized (BDCNM, DBCNM, TBNM, CNM, DCNM, BCNM). The rank order of their chronic cytotoxicity (72 h exposure) to Chinese hamster ovary (CHO) cells was DBNM > DBCNM > BNM > TBNM > BDCNM > BCNM > DCNM > CNM > TCNM. The rank order to induce genomic DNA damage in CHO cells was DBNM > BDCNM > TBNM > TCNM > BNM > DBCNM > BCNM > DCNM > CNM. The brominated nitromethanes were more cytotoxic and genotoxic than their chlorinated analogues. This research demonstrated the integration of the procedures for the analytical chemistry and analytical biology when working with limited amounts of sample. The halonitromethanes are potent mammalian cell cytotoxins and genotoxins and may pose a hazard to the public health and the environment.

A need exists for whole animal toxicity, mutagenesis, and carcinogenesis models that are alternative to the traditional rodent test models and that are economical, sensitive, and scientifically acceptable. Among small fish models, the Japanese medaka (Oryzias latipes) is preeminent for investigating effects of carcinogenic and/or toxic waterborne hazards to humans. The guppy (Poecilia reticulata), although less widely used, is valuable as a comparison species. Both species are easy to maintain and handle in the laboratory and there is a large body of background information on their responsiveness to a range of classes of carcinogens. There are considerable data on the occurrence of background diseases and on spontaneous neoplastic lesions, both of which occur relatively rarely. With few modifications, the medaka and guppy are amenable to carcinogenicity testing under the rigid standards established by the National Toxicology Program (NTP) for rodent tests. The advantages of the small fish in carcinogenesis studies are best realized in long-term studies that involve environmentally realistic exposures. Studies to identify chronic effects can be conducted in about 12 months, near the life span of medaka in our laboratory. Practically, 9-month studies are optimal but shorter study cycles and a variety of exposure/growout and initiation/promotion scenarios are available. Studies on 3 compounds tested in medaka under NTP protocols are under review and preliminary analysis indicates that chronic carcinogenicity bioassays with medaka, guppy, and potentially with other small fish species are feasible and scientifically valid.

In response to transmembrane potentials which are negative on the inner side of both the plasma and mitochondrial membranes, cationic dyes displaying appropriate structural features naturally accumulate in the cytosol and inside the mitochondria. Because enhanced mitochondrial membrane potential is a prevalent tumor cell phenotype, a number of cationic dyes preferentially accrue and are retained for longer periods in the mitochondria of tumor cells as compared to normal cells. The opportunities brought about by this phenomenon in chemo- and photochemotherapy of neoplastic diseases is highlighted by the observation that the phototoxic effects associated with some of the cationic photosensitizers known to accumulate in cell mitochondria are much more pronounced in tumor cells than in normal cells. However, the structural determinants of selective phototoxicity towards tumor cells are not well understood, and the lack of a robust model to describe the relationship between molecular structure and tumor selectivity has prevented mitochondrial targeting from becoming a more dependable therapeutic strategy. In this report we describe how the lipophilic/hydrophilic character of a series of cationic triarylmethane dyes affects the selectivity with which these photosensitizers mediate the destruction of tumor cells. Our results indicated that only the more hydrophilic triarylmethanes show tumor selectivity, presumably because these are the only dyes capable of staining energized mitochondria with a high degree of specificity. The partition of the more lipophilic dyes into a variety of extra-mitochondrial subcellular compartments occurs with comparable efficiencies in tumor and in normal cells, and this less specific subcellular localization precludes tumor selectivity from taking place.

The TOPKAT computer-based system for predicting chemical carcinogens was evaluated by determining its ability to predict the carcinogenicity of chemicals tested by the National Toxicology Program. TOPKAT was not effective in identifying potential rodent carcinogens and noncarcinogens in the data set analyzed. The chemicals in the TOPKAT database of known carcinogens and noncarcinogens that the software identifies as most "similar" to unknown chemicals are illustrated using six examples. These "similar" chemicals generally bear no apparent relationship to the chemical of interest with regard to metabolism or potential mechanism of carcinogenicity. Environ. Mol. Mutagen. 37:55-69, 2001 Published 2001 Wiley-Liss, Inc.

Chemicals such as cis-platinum, formaldehyde, chromate, copper, and certain arsenic compounds have been shown to produce DNA-protein cross-links in human in vitro cell systems at high doses, such as those in the cytotoxic range. Thus far there have only been a limited number of other chemicals evaluated for their ability to produce cross-links. The purpose of the work described here was to evaluate whether select industrial chemicals can form DNA-protein cross-links in human cells in vitro. We evaluated acetaldehyde, acrolein, diepoxybutane, paraformaldehyde, 2-furaldehyde, propionaldehyde, chloroacetaldehyde, sodium arsenite, and a deodorant tablet [Mega Blue; hazardous component listed as tris(hydroxymethyl)nitromethane]. Short- and long-term cytotoxicity was evaluated and used to select appropriate doses for in vitro testing. DNA-protein cross-linking was evaluated at no fewer than three doses and two cell lysate washing temperatures (45 and 65 degrees C) in Epstein-Barr virus (EBV) human Burkitt's lymphoma cells. The two washing temperatures were used to assess the heat stability of the DNA-protein cross-link, 2-Furaldehyde, acetaldehyde, and propionaldehyde produced statistically significant increases in DNA-protein cross-links at washing temperatures of 45 degrees C, but not 65 degrees C, and at or above concentrations of 5, 17.5, and 75 mM, respectively. Acrolein, diepoxybutane, paraformaldehyde, and Mega Blue produced statistically significant increases in DNA-protein cross-links washed at 45 and 65 degrees C at or above concentrations of 0.15 mM, 12.5 mM, 0.003%, and 0.1%, respectively. Sodium arsenite and chloroacetaldehyde did not produce significantly increased DNA-protein cross-links at either temperature nor at any dose tested. Excluding paraformaldehyde and 2-furaldehyde treatments, significant increases in DNA-protein cross-links were observed only at doses that resulted in complete cell death within 4 d following dosing. This work demonstrates that DNA-protein cross-links can be formed in vitro following exposure to a variety of industrial compounds and that most cross-links are formed at cytotoxic concentrations.

Male and female Long-Evans rats were housed in inhalation chambers and exposed to vapors of nitromethane (NM) at either 100 or 200 ppm. The animals were exposed 7 hr per day, 5 days per week for 2 years. Control groups of rats were also housed in a similar inhalation chamber, but NM was not introduced into the chamber. The animals were observed daily for signs of pharmacologic or toxicologic effect and body weights were recorded periodically. At the 2-year termination of the exposure period, clinical laboratory examinations (serum chemistry and hematology) were performed on selected animals and all surviving animals were sacrificed. All animals were necropsied and subjected to a thorough histopathologic examination. During the study there were no pharmacologic effects from exposure to NM at either 100 or 200 ppm. There was no effect on mortality on either sex at either exposure level. Body weights of male rats exposed to NM were not significantly different from those of control rats, but the body weights of female rats of both exposure groups were slightly less than their controls. There was no effect of exposure of rats of either sex to either level of NM on hematology. There were no clinically significant effects on serum chemistry. There were no effects of exposure to NM on organ weights. There were no significant differences in the nonneoplastic or neoplastic pathology related to exposure to NM.

Why is a low dose of toxic chemical nontoxic? What makes a larger dose of the same chemical toxic? Extensive work done to understand the mechanism of halomethane hepatotoxicity and its potentiation by chlorinated insecticide, chlordecone has resulted in the understanding of these basic tenets of toxicology. Studies suggest that ordinarily a small dose of halomethane causes limited liver injury which is accompanied by stimulated tissue repair enabling complete recovery from injury before manifestation. A large dose of halomethane becomes toxic due to suppressed tissue repair, which permits injury to progress in an unchecked fashion. Exposure to very low levels of chlordecone results in highly exaggerated toxicity of ordinarily nontoxic doses of halomethane because of suppressed hepatocellular regeneration and restoration, permitting the progression of liver injury ultimately resulting in liver failure and animal mortality. This concept is further supported by the observation that, while exposure to even high levels of phenobarbital and subsequent low nontoxic doses of halomethane results in greater level of initial liver injury, tissue repair is not completely suppressed; it is slightly postponed by 24 hr, but then much higher rate of tissue repair ensures and consequently enables the animals to completely recover from liver injury and survive. Thus, whether initiation of tissue repair processes occurs or not is the critical determinant in the ultimate manifestation of hepatotoxicity and its end result of either animal death or recovery and survival. Currently understood 'Mechanisms of toxicity' adequately explain only how toxic injury begins. These mechanisms do not permit us to predict the ultimate outcome of toxicity.(ABSTRACT TRUNCATED AT 250 WORDS)

In a previous study, the relationships between the chemical structure and the ability of 35 chlorinated aliphatic hydrocarbons to induce aneuploidy and toxicity in Aspergillus nidulans were analyzed. Quantitative structure-activity relationships (QSAR) were defined for each of the biological activities under study: ARR (the dose able to block mitotic growth), D37 (the dose with 37% of survival) and LEC (the lowest efficient concentration in aneuploidy induction). In this study, these QSAR equations were used to predict the toxic and genetic activity of a further six chemicals, not included in the previous data base: bromotrichloromethane, bromoform, bromochloromethane, bromodichloromethane, dibromochloromethane and dibromochlorofluoromethane. Their ARR, D37 and LEC values were measured, and were in agreement with the predicted values, with correlation coefficients around 0.99. Furthermore, the QSAR model, which had previously been developed to discriminate between aneugenic and inactive halogenated hydrocarbons, correctly predicted the aneugenic activity of five out of six methanes. These correct predictions confirmed the validity of our QSAR model, according to which the induction of aneuploidy in A. nidulans depends on both the electrophilic and steric properties of the chemicals, whereas toxicity mainly depends on steric factors.

The haploid strain 15B-II4 of Saccharomyces cerevisiae was used to study in an acute experiment the toxic and mutagenic effects of arylmethane dyes Victory Blue (C.I. 44040), Methyl Violet (C.I. 42535), Brilliant Green (C.I. 42040) and cancerogenic aminoazo dye Chrysoidine (C.I. 11270). High biological activity of all the dyes tested was found, based on such toxic effects as cell killing and growth inhibition. Also, it was shown that the dyes could increase the frequency of appearance of nuclear point mutations and cytoplasmic mutations of respiratory deficiency.

A major toxicological issue today is the possibility of unusual toxicity due to interaction of toxic chemicals upon environmental or occupational exposures to two or more chemicals, at ordinarily harmless levels individually. While some laboratory models exist for such interactions for the simplest case of only two chemicals, progress in this area has suffered for want of a model where the two interactants are individually nontoxic. One such model is available, where prior exposure to nontoxic levels of the pesticide Kepone (chlordecone) results in a 67-fold amplication of CCl4 lethality in rats. Extensive hepatotoxicity observed in this interaction is characterized by histopathological alterations, perturbation of related biochemical parameters and is followed by complete hepatic failure. This propensity for chlordecone to potentiate hepatotoxicity of halomethanes such as CCl4, CHCl3, and BrCCl3 has been a subject of intense study to unravel the underlying mechanism. Mechanisms such as induction of microsomal cytochrome P-450 by chlordecone and greater lipid peroxidation are inadequate to explain the remarkably powerful potentiation of halomethane toxicity. Compelling experimental evidence supports the hypothesis that hepatocellular division during early time points after the administration of CCl4 is an important determinant of the progression (or repair of it) of the liver injury and consequent destruction (or restoration) of the hepatolobular architecture and function. This paper advances a hypothesis for the mechanism of hepatotoxic and lethal effect of CCl4 as being primarily related to the accelerated progression of liver injury due to suppressed hepatocellular regeneration and hepatolobular restoration. This is in contrast to the widely accepted putative mechanism, one which invokes only bioactivation followed by runaway lipid peroxidation as the events determining the course of the progressive phase of liver injury. The concept being advanced in this paper accepts bioactivation (and perhaps lipid peroxidation) as the primary initiating events of cell injury, but maintains that they are not the determinants of the progressive phase of liver injury. The biological issue of whether the cells are incapacitated from regenerating is the determinant of the progression of liver injury, and therefore, the ultimate outcome of hepatotoxicity and lethality.