Long-term Chronic Exposure Research Articles

3-Nitrobenzanthrone promotes malignant transformation in human lung epithelial cells through the epiregulin-signaling pathway

Exposure to environmental and occupational contaminants leads to lung cancer. 3-Nitrobenzanthrone (3-nitro-7H-benz[de]anthracen-7-one, 3-NBA) is a potential carcinogen in ambient air or diesel particulate matter. Studies have revealed that short-term exposure to 3-NBA induces cell death, reactive oxygen species activation, and DNA adduct formation and damage. However, details of the mechanism by which chronic exposure to 3-NBA influences lung carcinogenesis remain largely unknown. In this study, human lung epithelial BEAS-2B cells were continuously exposed to 0-10-μM 3-NBA for 6 months. NanoString analysis was conducted to evaluate gene expression in the cells, revealing that 3-NBA-mediated transformation results in a distinct gene expression signature including carbon cancer metabolism, metastasis, and angiogenesis. Alterations in tumor-promoting genes such as EREG (epiregulin), SOX9, E-cadherin, TWIST, and IL-6 were involved in epithelial cell aggressiveness. Kaplan-Meier plotter analyses indicated that increased EREG and IL-6 expressions in early-stage lung cancer cells are correlated with poor survival. In vivo xenografts on 3-NBA-transformed cells exhibited prominent tumor formation and metastasis. EREG knockout cells exposed to 3-NBA for a short period exhibited high apoptosis and low colony formation. By contrast, overexpression of EREG in 3-NBA-transformed cells markedly activated the PI3K/AKT and MEK/ERK signaling pathways, resulting in tumorigenicity. Furthermore, elevated IL-6 and EREG expressions synergistically led to STAT3 signaling activation, resulting in clonogenic cell survival and migration. Taken together, chronic exposure of human lung epithelial cells to 3-NBA leads to malignant transformation, in which the EREG signaling pathway plays a pivotal mediating role. • Short-term exposure of lung epithelial cells to 3-NBA can lead to ROS production and cell apoptosis. • Long-term chronic exposure to 3-NBA upregulates the levels of tumor-promoting genes such as EREG and IL-6. • Increased EREG expression in 3-NBA-transformed cells markedly contributes to tumorigenesis through PI3K/AKT and MEK/ERK activation and synergistically enhances the IL-6/STAT3 signaling pathway, which promotes tumorigenicity.

Interpreting Risk from Sunscreens in the Marine Environment.

Recent and pending bans in specific jurisdictions of some organic ultraviolet (UV) filters have resulted in significant concern and controversy over the potential impacts of these contaminants in the marine environment. Organic UV filters have been quantified in the aquatic environment as contaminants in water, sediments, and the tissues of aquatic organisms. The limited available laboratory studies on the toxicity of UV filters to keystone marine species such as reef-building corals describe a wide variety of impacts, from significant acute effects to no observed effects. However, interpretation of results is complicated by differences in methodology, and exposures to single agents in vitro may not reflect the effects of longer exposure to finished sunscreens containing UV filters in combination with numerous other chemicals. Relatively short-term observations of laboratory effects thus may not translate to real-life field conditions, where organisms may be subject to the effects of long-term chronic exposure to UV filters as well as other environmental contaminants and stressors. The lack of current understanding of the full impacts of UV filters, both in the laboratory and in the environment, represents a significant challenge in interpreting the environmental risk associated with the widespread use of sunscreens.

Rare cases of severe life-threatening lead poisoning due to accident or chronic occupational exposure to lead and manganese: Diagnosis, treatment, and prognosis

Chronic long-term, low-dose environmental and occupational exposure to lead (Pb) has been extensively studied in large cohorts worldwide among general populations, miners, smelters, or battery workers. However, studies on severe life-threatening Pb poisoning due to accidental or chronic occupational exposure to Pb and manganese (Mn) were rarely reported. We present one case of acute severe Pb poisoning and compare it with another severe chronic occupational exposure case involving Pb and Mn. A 27-year-old woman mistakenly took a large quantity of pure Pb powder as an herbal remedy; she developed abdominal colic, severe nausea, vomiting, fatigue, and cutaneous and sclera icterus. Laboratory tests showed her blood lead level (BLL) of 173.5 µg dL-1 and urinary lead level (ULL) of 1240 µg dL-1. The patient was diagnosed with acute Pb poisoning and acute liver failure. In another chronic exposure case, a 56-year-old man worked in a Pb and Mn smelting factory for 15 years. He was brought to the emergency room with severe nausea, vomiting, and paroxysmal abdominal colic, which was intolerable during the onset of pain. His BLL was 64.8 µg dL-1 and ULL was 38 µg dL-1, but his blood and urinary Mn levels were normal. The patient was diagnosed with chronic Pb poisoning. Both patients received chelation therapy with calcium disodium ethylene-diamine-tetraacetate (CaNa2EDTA). The woman with acute severe Pb intoxication recovered well and was discharged from the hospital after treatment, and the man who survived severe Pb poisoning was diagnosed with lung cancer. Clinical manifestations of acute and chronic severe Pb poisoning are different. Chelation therapy with CaNa2EDTA is proven to be an effective life-saving therapy in both cases by reducing BLL. Occupational exposure to both Pb and Mn does not appear to increase Mn neurotoxicity; however, the probability that co-exposure to Mn may increase Pb toxicity in the same patient cannot be excluded.

Agrochemicals in freshwater systems and their potential as endocrine disrupting chemicals: A South African context.

South Africa is the largest agrochemical user in sub-Saharan Africa, with over 3000 registered pesticide products. Although they reduce crop losses, these chemicals reach non-target aquatic environments via leaching, spray drift or run-off. In this review, attention is paid to legacy and current-use pesticides reported in literature for the freshwater environment of South Africa and to the extent these are linked to endocrine disruption. Although banned, residues of many legacy organochlorine pesticides (endosulfan and dichlorodiphenyltrichloroethane (DDT)) are still detected in South African watercourses and wildlife. Several current-use pesticides (triazine herbicides, glyphosate-based herbicides, 2,4-dichlorophenoxyacetic acid (2,4-D) and chlorpyrifos) have also been reported. Agrochemicals can interfere with normal hormone function of non-target organism leading to various endocrine disrupting (ED) effects: intersex, reduced spermatogenesis, asymmetric urogenital papillae, testicular lesions and infertile eggs. Although studies investigating the occurrence of agrochemicals and/or ED effects in freshwater aquatic environments in South Africa have increased, few studies determined both the levels of agricultural pesticides present and associated ED effects. The majority of studies conducted are either laboratory-based employing in vitro or in vivo bioassays to determine ED effects of agrochemicals or studies that investigate environmental concentrations of pesticides. However, a combined approach of bioassays and chemical screening will provide a more comprehensive overview of agrochemical pollution of water systems in South Africa and the risks associated with long-term chronic exposure.

Exposure to organophosphorus insecticides and increased risks of health and cancer in US women

Results of this paper provide evidence that chronic long-term exposure to organophosphorus insecticides poses a significantly higher health risk for US women than for men, based on dialkylphosphate biomarker data from NHANES cycles 2003-2012. The risk of cardiovascular disease for female non-smokers aged 60-85 years in the highest dimethylthiophosphate (DMTP) urinary concentration quartile is 3.0 (odds ratio, OD = 3.0, 95%CI 1.4-6.4) times higher than that in the lowest quartile. Women with higher urinary DMTP concentrations also have significantly higher risk of asthma at the ages 6-39 years and an apparently higher risk of chronic bronchitis at the ages 60-85. Overall cancer risk is significantly higher for female non-smokers aged 60-85 years in the higher urinary DMTP quartiles (OD = 2.7, 95% CI 1.3-5.9). Increasing risks of breast cancer for female smokers and prostate cancer for male smokers aged 60-85 years with higher exposure to organophosphorus insecticides in the US are also significant.

Involvement of overweight and lipid metabolism in the development of pulmonary hypertension under conditions of chronic intermittent hypoxia.

There is growing evidence that exposure to hypoxia, regardless of the source, elicits several metabolic responses in individuals. These responses are constitutive and are usually observed under hypoxia but vary according to the type of exposure. The aim of this review was to describe the involvement of obesity and lipid metabolism in the development of high-altitude pulmonary hypertension and in the development of acute mountain sickness under chronic intermittent hypoxia. Overweight or obesity, which are common in individuals with long-term chronic intermittent hypoxia exposure (high-altitude miners, shift workers, and soldiers), are thought to play a major role in the development of acute mountain sickness and high-altitude pulmonary hypertension. This association may be rooted in the interactions between obesity-related metabolic and physical alterations, such as increased waist circumference and neck circumference, among others, which lead to critical ventilation impairments; these impairments aggravate hypoxemia at high altitude, thereby triggering high-altitude diseases. Overweight and obesity are strongly associated with higher mean pulmonary artery pressure in the context of long-term chronic intermittent hypoxia. Remarkably, de novo synthesis of triglycerides by the sterol regulatory element-binding protein-1c pathway has been demonstrated, mainly due to the upregulation of stearoyl-CoA desaturase-1, which is also associated with the same outcomes. Therefore, overweight, obesity, and other metabolic conditions may hinder proper acclimatization. The involved mechanisms include respiratory impairment, alteration of the nitric oxide pathways, inflammatory status, reactive oxygen species imbalance, and other metabolic changes; however, further studies are required.

Haemophilus influenzae persists in biofilm communities in a smoke-exposed ferret model of COPD.

RationaleNon-typeable Haemophilus influenzae (NTHi) is a common inhabitant of the human nasopharynx and upper airways that can cause opportunistic infections of the airway mucosa including bronchopulmonary infections in patients with chronic obstructive pulmonary disease (COPD). It is clear that opportunistic infections contribute significantly to inflammatory exacerbations of COPD; however, there remains much to be learned regarding specific host and microbial determinants of persistence and/or clearance in this context.MethodsIn this study, we used a recently described ferret model for COPD, in which animals undergo chronic long-term exposure to cigarette smoke, to define host–pathogen interactions during COPD-related NTHi infections.ResultsNTHi bacteria colonised the lungs of smoke-exposed animals to a greater extent than controls, and elicited acute host inflammation and neutrophilic influx and activation, along with a significant increase in airway resistance and a decrease in inspiratory capacity consistent with inflammatory exacerbation; notably, these findings were not observed in air-exposed control animals. NTHi bacteria persisted within multicellular biofilm communities within the airway lumen, as evidenced by immunofluorescent detection of bacterial aggregates encased within a sialylated matrix as is typical of NTHi biofilms and differential bacterial gene expression consistent with the biofilm mode of growth.ConclusionsBased on these results, we conclude that acute infection with NTHi initiates inflammatory exacerbation of COPD disease. The data also support the widely held hypothesis that NTHi bacteria persist within multicellular biofilm communities in the lungs of patients with COPD.

Behavioral plasticity and gene regulation in the brain during an intermittent ethanol exposure in adult zebrafish population

Ethanol consumption is correlated with different neurobiological and behavioral impairments. Acute and chronic exposure to this drug is associated with alterations in the regulation of the mesolimbic dopaminergic system as well as with transcriptional modulation of other receptors in the central nervous system and can unleash seeking behavior or behavioral adaptations and phenotypes such as loss of control, dependence and tolerance. In the present work, we characterized the chronological effects of acute and chronic intermittent exposure to ethanol (1% v/v) in an adult zebrafish population (Danio rerio). During sixteen days of ethanol exposure, we associated the neuromodulation of target genes (drd1, drd2, gabra2a, gabbr1a, gabbr1b) in the central nervous system with behavioral parameters, assessed by social preference, antipredatory capacity and anxiety-like analysis. Transcriptional and behavioral data were collected in days 0, 1, 4, 8, 12 and 16, after ethanol exposure. In days 1 and 4, ethanol exposure increased exploratory behavior regardless of the risk involved (less time spent close to conspecifics and lower avoidance reaction to predator). Along with the reduction of drd2, grin1a and gabra2a transcription seen in the same days, these results suggest an anxiolytic effect of acute ethanol exposure. Interestingly, in days 8, 12 and 16, an attenuation of the behavioral effects was observed. The social preference, antipredatory behavior, perception and exploration parameters were reconstituted. This behavioral re-establishment, accompanied by the increase in drd1, drd2 and gabbr1a transcription in the 8th day could be an indicative of an adaptation to chronic exposure to ethanol. The modulation of drd2 gene combined with the behavioral characterization observed in the study suggests this signalling pathway as a key participant in the phenotypic outcomes of a long-term chronic exposure to ethanol. Lastly, our results reaffirm the ethanol deleterious impacts in perception, ability to respond to adverse stimuli and in anxiety-like behavior.

BACKGROUND RADIATION LEVELS NEAR A MINERAL SAND MINING FACTORY IN SRI LANKA: CORRELATION OF RADIATION MEASUREMENTS WITH MICRONUCLEI FREQUENCY.

Lanka Mineral Sands Ltd (LMS) is a company operating in Pulmoddai, Sri Lanka, mining for rare earth minerals along with monazite which contains thorium that emits ionizing gamma and alpha radiation on decay. The objective of the study was to determine the background radiation levels and selected radionuclides and then to correlate these levels with the frequency of micronuclei (MN) among persons residing in the vicinity of LMS. A cross-sectional study was conducted among persons of both sexes between 35 and 45years of age residing in the vicinity of LMS. Background radiation measurements were obtained by a survey metre, and gamma spectrometry was done on soil samples. Five millilitres of venous blood was drawn for cytokinesis-blocked MN assay. Background radiation levels measured by the survey metre; 232Th, 226Ra and 210Pb mass activities in soil were highest in the samples collected from the LMS. The background radiation measurements positively correlated with MN frequency although the magnitude of the correlation was small (r=0.176, p=0.04). This implies that chronic long-term exposure to low-dose radiation may result in genotoxicity. Prospective large-scale studies are recommended to evaluate the long-term effect of exposure to low-dose radiation at Pulmoddai.

Air pollution and health: Evidence from epidemiological studies and population impact

Outdoor air pollution —in particular particulate matter, nitrogen dioxide and ozone— can exert its effects on health after acute (short-term) and chronic (long-term) exposures. Short-term exposures increase the probability of the onset of acute diseases within a few days, such as myocardial infarction or stroke, or even death in the case of susceptible individuals. Long-term exposures are associated with decreased survival and incidence of several non-communicable diseases, including cardiorespiratory conditions and lung cancer. In Europe, the large ESCAPE project (European Study of Cohorts for Air Pollution Effects — www.escapeproject.eu) evaluated the chronic effects of air pollution in the cohorts of adult subjects. The results of ESCAPE show an association between chronic exposure to air pollutants and natural mortality, cardiovascular events, lung, brain, breast and digestive tract cancer. The recent joint statement of the European Respiratory Society and the American Respiratory Society clarifies the wide spectrum of adverse effects of pollution, including “new” diseases such as neurological and metabolic syndrome previously not studied. The estimates by the Global Burden of Disease provide nowadays indications that air pollution causes illness and mortality, just after diet, smoking, hypertension and diabetes: 4.2 million premature deaths a year worldwide. Ischemic heart disease, stroke, chronic obstructive pulmonary disease, acute lower respiratory infections are the main conditions associated with air-pollution–related mortality.

Dietary exposure to neonicotinoid insecticides and health risks in the Chinese general population through two consecutive total diet studies

Neonicotinoid insecticides are ubiquitous in food and the environment due to their wide use. Growing evidence suggests the adverse effects of neonicotinoids in many species, including mammals. Some studies have reported the urinary concentrations of neonicotinoids in human biological monitoring, but the potential risks of neonicotinoids on human health based on long-term chronic exposure studies in any general population have been rarely tackled. In this study, the dietary exposure to neonicotinoids of the Chinese adult population was studied on the basis of composite dietary samples collected from the 5th (2009–2012) and 6th (2015–2018) Chinese total diet studies (TDS). Residue levels of ten neonicotinoids were determined in 528 composite dietary samples from 24 provinces in China. Most of the samples (53.3% and 70.5% in the 5th and 6th TDS, respectively) that we analyzed contained the multi-residue of neonicotinoids. Imidacloprid and acetamiprid were the most frequently detected neonicotinoids, and thiamethoxam and clothianidin were increasingly used and found in the 6th TDS. The estimated daily intake (EDI) for total neonicotinoids was calculated to evaluate health risk of the Chinese adult population based on a relative potency factor assessment method. The mean EDIs of total neonicotinoids in the 5th and 6th TDS respectively reached 598.95 and 710.38 ng/kg bw per day. Although the mean EDIs of total neonics in 6th TDS was relatively higher than that in 5th TDS, no statistical difference was observed (p > 0.05). Vegetables were the main source of dietary exposure, but exposure via cereals and beverages and water must also be addressed in China. Although the average exposure for total neonicotinoids was much lower than the current chronic reference dose (57 μg/kg bw per day), the dietary exposure risks of a general population for total neonicotinoids should not be overlooked due to the ubiquity of neonicotinoids in food and the environment.

Chronic long-term exposure to airborne dioxin at a national level, and breast cancer risk in a nested case-control study in the French E3N cohort

Chronic long-term exposure to airborne dioxin at a national level, and breast cancer risk in a nested case-control study in the French E3N cohort

Chronic long-term exposure to cadmium air pollution and breast cancer risk in the French E3N cohort

Chronic long-term exposure to cadmium air pollution and breast cancer risk in the French E3N cohort

Effects of microplastic particles and leaching additive on the life history and morphology of Daphnia magna

Plastic waste is continuously introduced not only into marine, but also freshwater environments, where it fragments into microplastics. Organisms may be affected by the particles themselves due to ingestion and indirectly via incorporated additives such as plasticizers, since these substances have the ability to leach out of the polymer matrix. Although it has been indicated that the likelihood of additives leaching out into the gut lumen of organisms exposed to microplastics is low, studies distinguishing between the effects of the synthetic polymer itself and incorporated additives of the same polymer are scarce. Since this is obligatory for risk assessment, we analyzed the chronic effects of flexible polyvinylchloride (PVC), a widely used polymer, containing the plasticizer diisononylphthalate (DiNP) on morphology and life history of the freshwater crustacean Daphnia magna and compared these effects with the effects of rigid PVC, lacking DiNP, as well as a glass bead control. After up to 31 days of exposure, rigid PVC and glass beads did not affect body length and relative tail spine length of D. magna, whereas flexible PVC led to an increased body length and a reduced number of offspring. None of the treatments increased the mortality significantly. We were able to show that 2.67μg/L DiNP leached out of the flexible PVC into the surrounding medium using GC-MS. Yet, we were not able to measure leachate inside the gut lumen of D. magna. The effects emerged towards the end of the experiment, due to the time dependent process of leaching. Therefore, the results highlight the relevance of long-term chronic exposure experiments, especially as leaching of additives takes time. Further, our study shows the importance to distinguish between microplastics containing leachable additives and the raw polymer in ecotoxicological testing.

441 Overlapping functions of PARP1 and p63 in cutaneous squamous cell carcinoma

Cutaneous SCC (cSCC) is the second most frequent type of human malignancy worldwide primarily caused by chronic long-term sun exposure. Somatic mutations in the tumor suppressor gene TP53 have been shown to occur frequently in aged sun-exposed skin and to favor clonal expansion. In contrast, p63, a p53 family member, is rarely mutated in cancer and rather is overexpressed and/or amplified in squamous cell carcinomas. Several studies have shown that p63 overexpression leads to higher proliferation rates and an increased ability to contrast differentiation, cellular senescence and apoptosis. Using a proteomic screening, we identified PARP1 as a p63 protein partner. PARP1 is an enzyme that add PAR moieties onto target proteins and regulates different cellular processes including DNA repair and transcriptional regulation. Furthermore, it is a known therapeutic target for several tumor types including breast and ovarian cancer. Similar to p63, PARP1 is abundantly expressed in a large cohort of cSCC samples as compared to normal skin and is upregulated in cSCC cell lines. To investigate whether PARP1 and p63 coordinately regulate global patterns of gene expression, either PARP1 or p63 were knocked down in SCC cells followed by RNA-seq. Correlation analyses of the transcriptomic profile under basal condition, showed that a large proportion of genes affected by the depletion of one factor was similarly affected by the depletion of the other, causing a highly significant coordinated perturbation in transcription. Furthermore, most of the downregulated genes in PARP1 and p63 depleted cells are involved specifically in cell cycle progression and mitosis. Taken together these data reveal a previously unrecognized functional interaction between p63 and PARP1 in regulating transcription of a large set of cell cycle genes during early stages of skin tumorigenesis, potentially opening a window for novel therapeutic intervention.

Simvastatin affect the expression of detoxification-related genes and enzymes in Daphnia magna and alter its life history parameters

Simvastatin (SV), as an hypocholesterolaemic drug, has been detected in various aquatic environment. However, limited information is available on the effects of SV on freshwater invertebrates. In the present study, we investigated the toxic effects of SV on Daphnia. magna (D. magna) through measuring the physiological changes (e.g., survival, growth rate, and reproduction) in a 21-d chronic toxicity test We also determined the expression of seven detoxification and reproduction-related genes (i.e. HR96, P-gp, CYP360A8, GST, CYP314, EcR and Vtg) and several enzymes (i.e. APND, ERND, GST and CAT) in a acute test (24 h). Results showed that high concentration (e.g. 50 μg L−1) of SV for short time exposure (e.g. 24 h) significantly induced the expression of HR96 and P-gp (e.g. up to 2.5 folds)and enzymes (e.g. increasing 4.0 folds for ERND and GST activity) in D. magna.. The long-term chronic exposure (21 days) may cause the changes of life history parameters such as decreasing total egg production number per individual and intrinsic growth rates etc. SV may act as a potential endocrine disruptor to D. magna and the reproduction parameters were more sensitive endpoints than the survival and growth for evaluating SV exposure.

Conserved Microbial Toxicity Responses for Acute and Chronic Silver Nanoparticle Treatments in Wetland Mesocosms.

Most studies of bacterial exposure to environmental contaminants focus on acute treatments; however, the impacts of single, high-dose exposures on microbial communities may not readily be extended to the more likely scenario of chronic, low-dose contaminant exposures. Here, in a year-long, wetland mesocosm experiment, we compared microbial community responses to pulse (single 450 mg dose of silver) and chronic (weekly 8.7 mg doses of silver for 1 year) silver nanoparticle (Ag0 NP) treatments, as well as a chronic treatment of "aged" sulfidized silver nanoparticles (Ag2S NPs). While mesocosms exposed to Ag2S NPs never differed significantly from the controls, both Ag0 NP treatments exhibited reduced microbial diversity and altered community composition; however, the effects differed in timing, duration, and magnitude. Microbial community-level impacts in the acute Ag0 NP treatment were apparent only within the first weeks and then converged on the control mesocosm composition, while chronic exposure effects were observed several months after exposures began, likely due to interactive effects of nanoparticle toxicity and winter environmental conditions. Notably, there was a high level of overlap in the taxa which exhibited significant declines (>10×) in both treatments, suggesting a conserved toxicity response for both pulse and chronic exposures. Thus, this research suggests that complex, but short-term, acute toxicological studies may provide critical, cost-effective insights into identifying microbial taxa sensitive to long-term chronic exposures to Ag NPs.

Contributing Roles of CYP2E1 and Other Cytochrome P450 Isoforms in Alcohol-Related Tissue Injury and Carcinogenesis.

The purpose of this review is to briefly summarize the roles of alcohol (ethanol) and related compounds in promoting cancer and inflammatory injuryin many tissues. Long-term chronic heavy alcohol exposure is known to increase the chances of inflammation, oxidative DNA damage,and cancer development in many organs. The rates of alcohol-mediated organ damage and cancer risks are significantly elevated in the presence of co-morbidity factors such as poor nutrition, unhealthy diets, smoking, infection with bacteria orviruses, and exposure to pro-carcinogens. Chronic ingestion of alcohol and its metabolite acetaldehyde may initiate and/or promote the development of cancer in the liver, oral cavity, esophagus, stomach, gastrointestinal tract, pancreas, prostate, and female breast. In this chapter, we summarize the important roles of ethanol/acetaldehyde in promoting inflammatory injuryand carcinogenesis in several tissues. We also review the updated roles of the ethanol-inducible cytochrome P450-2E1 (CYP2E1) and other cytochrome P450 isozymes in the metabolism of various potentially toxic substrates, and consequent toxicities, including carcinogenesis in different tissues. We also briefly describe the potential implications of endogenous ethanol produced by gut bacteria, as frequently observed in the experimental models and patients of nonalcoholic fatty liver disease, in promoting DNA mutation and cancer development in the liver and other tissues, including the gastrointestinal tract.

δ13C, δ15N, N concentration, C/N, and Ca/Al of Pinus densiflora foliage in Korean cities of different precipitation pH and atmospheric NO2 and SO2 levels

Forest decline due to atmospheric pollution is a consequence of long-term chronic exposure, and thus chemical proxies that are sensitive to the pollution are helpful in estimating the impact of atmospheric pollution on forest health. In this study, we explored changes in the isotopic and elemental compositions of needles of red pine (Pinus densiflora) with varying precipitation pH and concentrations of NO2 and SO2 across 18 cities to identify isotopic and chemical signatures that are sensitive to acid deposition. The cities had different intensity of traffic and industrial activities, and thus were expected to have varying levels of atmospheric pollution. The pine needles were analyzed for carbon (δ13C) and nitrogen (δ15N) isotope ratios, N concentrations, C-to-N ratio (C/N), and calcium-to-aluminum ratio (Ca/Al). Analysis of variance, simple linear correlation, and redundancy analysis were used to investigate the variations in foliar chemistry with atmospheric variables. Neither NO2 nor SO2 concentration was correlated with foliar δ13C, δ15N, N concentration, and C/N; whereas precipitation pH was correlated with the foliar parameters, suggesting that foliar chemistry is more sensitive to total acidifying materials than to individual pollutant. The foliar δ13C decreased with lowered precipitation pH, reflecting the increased 13C-depleted CO2 that was co-emitted with acidifying materials from fossil fuel combustion. Foliar δ15N decreased and foliar N concentration increased with lowered precipitation pH, indicating tree uptake of 15N-depleted N from acid deposition. Accordingly, the C/N ratio also decreased with lowered precipitation pH. However, there was no relationship between foliar Ca/Al and precipitation pH; rather. Our result suggests that δ13C, δ15N, N concentration, and C/N of pine needle samples are associated with the level of precipitation pH and thus pine needles could be used as bio-indicators of the impacts of total acidifying pollutants on forest.

Characterization of the proteome and lipidome profiles of human lung cells after low dose and chronic exposure to multiwalled carbon nanotubes

The effects of long-term chronic exposure of human lung cells to multi-walled carbon nanotubes (MWCNT) and their impact upon cellular proteins and lipids were investigated. Since the lung is the major target organ, an in vitro normal bronchial epithelial cell line model was used. Additionally, to better mimic exposure to manufactured nanomaterials at occupational settings, cells were continuously exposed to two non-toxic and low doses of a MWCNT for 13-weeks. MWCNT-treatment increased ROS levels in cells without increasing oxidative DNA damage and resulted in differential expression of multiple anti- and pro-apoptotic proteins. The proteomic analysis of the MWCNT-exposed cells showed that among more than 5000 identified proteins; more than 200 were differentially expressed in the treated cells. Functional analyses revealed association of these differentially regulated proteins to cellular processes such as cell death and survival, cellular assembly, and organization. Similarly, shotgun lipidomic profiling revealed accumulation of multiple lipid classes. Our results indicate that long-term MWCNT-exposure of human normal lung cells at occupationally relevant low-doses may alter both the proteome and the lipidome profiles of the target epithelial cells in the lung.