Adam19 Deficiency Impacts Pulmonary Function: Human GWAS Follow-up in Mouse.

Purpose: Over 550 loci have been associated with human pulmonary function in genome-wide association studies (GWAS); however, the causal role of most remains uncertain. Single nucleotide polymorphisms in a disintegrin and metalloprotease domain 19 (ADAM19) are consistently related to pulmonary function in GWAS. Thus, we used a mouse model to investigate the causal link between Adam19 and pulmonary function. Methods: We created an Adam19 knockout (KO) mouse model and validated the gene targeting using RNA-Seq and RT-qPCR. Contrary to prior publications, the KO was not neonatal lethal. Thus, we phenotyped the Adam19 KO. Results: KO mice had lower body weight and shorter tibial length than wild type (WT). Dual-energy X-ray Absorptiometry indicated lower soft weight, fat weight, and bone mineral content in KO mice. In lung function analyses using flexiVent, compared to WT, Adam19 KO had decreased baseline respiratory system elastance, minute work of breathing, tissue damping, tissue elastance, and forced expiratory flow at 50% forced vital capacity but higher FEV0.1 and FVC. Adam19 KO had attenuated tissue damping and tissue elastance in response to methacholine following LPS exposure. Adam19 KO also exhibited attenuated neutrophil extravasation into the airway after LPS administration compared to WT. RNA-Seq analysis of KO and WT lungs identified several differentially expressed genes (Cd300lg, Kpna2, and Pttg1) implicated in lung biology and pathogenesis. Gene set enrichment analysis identified negative enrichment for TNF pathways. Conclusion: Our murine findings support a causal role of ADAM19, implicated in human GWAS, in regulating pulmonary function.

Occurrence and spatial distribution of individual polychlorinated biphenyl congeners in residential soils from East Chicago, southwest Lake Michigan.

We report individual polychlorinated biphenyl congeners and the sum of all congeners (ΣPCB) in residential soils of East Chicago, Indiana. ΣPCB in soils ranged from 20 to 1700 ng/g dry weight (DW), with a geometric mean of 120 ng/g DW. These values are significantly higher than other locations, but similar or lower to locations nearby well-known PCB contamination sites. No PCB spatial distribution pattern was observed. PCB concentrations increase with total organic carbon in the soils and proximity to Indiana Harbor and Ship Canal (IHSC), where sediments are contaminated with PCBs. Most samples are similar in their PCB distribution and Aroclor 1254 yielded the highest similarity to all the samples. A fifth of the samples highly resemble other PCB profiles such as EPA background and Cedar Rapids Iowa soils, and volatilization from Lake Michigan, whereas volatilization from IHSC could not explain the PCBs found in soils. IHSC was expected to be the main source of PCBs in the nearby soils. It is possible that soils are impacted by variety of known and unknown sources, including volatilization from Lake Michigan, resulting in a regional PCB signal. Although PCB concentrations are higher than other locations, samples were below the current US EPA non-cancer residential soil level remediation goal for dioxin TEQ.

Time course of pulmonary inflammation and trace element biodistribution during and after sub-acute inhalation exposure to copper oxide nanoparticles in a murine model.

BACKGROUND: It has been shown that copper oxide nanoparticles (CuO NPs) induce pulmonary toxicity after acute or sub-acute inhalation exposures. However, little is known about the biodistribution and elimination kinetics of inhaled CuO NPs from the respiratory tract. The purposes of this study were to observe the kinetics of pulmonary inflammation during and after CuO NP sub-acute inhalation exposure and to investigate copper (Cu) biodistribution and clearance rate from the exposure site and homeostasis of selected trace elements in secondary organs of BALB/c mice. RESULTS: Sub-acute inhalation exposure to CuO NPs led to pulmonary inflammation represented by increases in lactate dehydrogenase, total cell counts, neutrophils, macrophages, inflammatory cytokines, iron levels in bronchoalveolar lavage (BAL) fluid, and lung weight changes. Dosimetry analysis in lung tissues and BAL fluid showed Cu concentration increased steadily during exposure and gradually declined after exposure. Cu elimination from the lung showed first-order kinetics with a half-life of 6.5 days. Total Cu levels were significantly increased in whole blood and heart indicating that inhaled Cu could be translocated into the bloodstream and heart tissue, and potentially have adverse effects on the kidneys and spleen as there were significant changes in the weights of these organs; increase in the kidneys and decrease in the spleen. Furthermore, concentrations of selenium in kidneys and iron in spleen were decreased, pointing to disruption of trace element homeostasis. CONCLUSIONS: Sub-acute inhalation exposure of CuO NPs induced pulmonary inflammation, which was correlated to Cu concentrations in the lungs and started to resolve once exposure ended. Dosimetry analysis showed that Cu in the lungs was translocated into the bloodstream and heart tissue. Secondary organs affected by CuO NPs exposure were kidneys and spleen as they showed the disruption of trace element homeostasis and organ weight changes.

In vitro and in silico bioaccessibility of urban dusts contaminated by multiple legacy sources of lead (Pb).

Lead contamination from gasoline, paint, pesticides, and smelting have unique chemical structures. Recent investigations into Pb speciation in urban soils and dusts from multiple sources have revealed emerging forms which differ from the initial sources. This results from reactions with soil constituents leading to transformation to new forms for which the bioaccessibilities remain uninvestigated. We investigated the in vitro and in silico bioaccessibility of these emerging forms in three physiologically relevant milieux: artificial lysosomal fluid (ALF), simulated epithelial lung fluid (SELF), and simulated gastric fluid (SGF). Species were validated using extended X-ray absorption fine structure spectroscopy. Results highlight diverse bioaccessibilities which are form and compartmentally-dependent. In ALF the bioaccessibility trend was humate-bound Pb (86%) > hydrocerussite (79%) > Fe oxide-bound Pb (47%) > galena (10%) > pyromorphite (4%) > Mn oxide-bound Pb (2%). Humate-bound Pb, hydrocerussite, Fe and Mn oxide-bound Pb were 100% bioaccessible in SGF while pyromorphite and galena were 26%, and 8%, respectively. Bioaccessibility in SELF was very low (< 1%) and significantly lower than ALF and SGF (p < 0.001). In silico bioaccessibilities modeled using equilibrium solubilities in extraction solutions were in good agreement with empirical measurements. These emerging forms of Pb have a wide range of bioaccessibilities that can influence their toxicity and impact on human health.

Validation of blood arsenic and manganese assessment from archived clotted erythrocyte fraction in an urban cohort of mother-child dyads.

Exposure to arsenic (As) and manganese (Mn) from contaminated food, drinking water and dust are linked to a host of adverse health effects. The recent discovery of unmonitored community exposures to hazardous levels of metals, as seen in the Flint Water Crisis and East Chicago, have demonstrated a need for novel biomonitoring methods utilizing samples other than whole blood. Here, we present a method utilizing clotted erythrocyte fraction samples, a blood component commonly archived in biorepositories, to predict whole blood levels of As and Mn. This method would allow for innovative retrospective assessments of environmental exposures in previously unused samples. Whole blood and clotted erythrocyte fraction samples were simultaneously collected from 84 participants in the Airborne Exposure to Semivolatile Organic Pollutants (AESOP) cohort study of mother-child dyads in East Chicago. Clotted erythrocyte fraction samples were prepared by alkaline dilution and subsequently analyzed using inductively coupled plasma-mass spectrometry. A strong linear relationship was observed between whole blood and clotted erythrocyte fraction with Pearson correlation coefficients (r, p < 0.001) of 0.74, and 0.82 for As and Mn, respectively. Modeled whole blood Mn levels predicted from clotted erythrocyte fractions evaluated at a test threshold representing the NHANES median of 9.7 µg/L, were found to have diagnostic sensitivity of 88% and specificity of 71%. Clotted erythrocyte partitioning of As was tested on a wide range of oral gavage doses using a rat model. Results from this investigation demonstrate clotted erythrocyte fraction samples are a viable alternative biological sample for retrospective public health surveillance of environmental exposure to As and Mn.

Lead (Pb) concentrations and speciation in residential soils from an urban community impacted by multiple legacy sources.

In many urban areas, elevated soil lead (Pb) concentrations are indicators of community-level Pb exposure. Here, we examine the spatial distribution and speciation of legacy soil Pb contamination in East Chicago, Ind., an industrial center with a wide range of Pb sources including a former lead smelter. In situ X-ray fluorescence spectroscopy (n = 358) revealed widespread soil Pb contamination above the Environmental Protection Agency regulatory limit for soils. This soil contamination was heterogenous across all neighborhoods, and mostly uncorrelated with distance from the former smelting site. Soil Pb levels increased with decreasing median household income in East Chicago's nine neighborhoods (r = -0.73, p = 0.03). Extended X-ray absorption fine structure spectroscopy (n = 44) indicated that the soil Pb was primarily adsorbed to iron and manganese oxides or humic acids, and as Pb hydroxycarbonate regardless of contamination levels. Crystalline insoluble forms of Pb, like pyromorphite, were not detected in significant concentrations. Thus, the unique chemical forms of potential Pb sources to soil, such as paint, ore and slag are not persistent and instead are extensively repartitioned into acid-soluble forms of Pb with greater bioavailability. These findings have implications for remediation efforts and human health as blood Pb levels in this community are significantly elevated.

Retrospective blood lead assessment from archived clotted erythrocyte fraction in a cohort of lead-exposed mother-child dyads.

Exposure to lead (Pb) is linked to a host of adverse health effects. Recent revelations of unmonitored exposures to hazardous levels of lead as seen in the Flint Water Crisis have manifested a need for evaluating biological samples, other than regularly prepared whole blood, for biomonitoring of lead exposure. Here, we present a method utilizing clotted erythrocyte fraction samples, which are commonly archived along with serum (or plasma) in biorepositories, to predict whole blood lead levels to allow for retrospective assessments of environmental exposure to lead. Whole blood and clotted erythrocyte fraction samples were collected from 91 participants in the Airborne Exposure to Semivolatile Organic Pollutants (AESOP) cohort study of mother-child dyads. Clotted erythrocyte fraction samples were prepared either using microwave assisted acid digestion or alkaline dilution and subsequently analyzed for lead using Inductively-couple plasma-mass spectrometry (ICP-MS). Simultaneously withdrawn whole blood samples were also analyzed. A strong linear relationship was observed between lead in whole blood and clotted erythrocyte fraction with Pearson correlation coefficients (r) of 0.90, and 0.89 (p < 0.001) for acid digestion and alkaline dilution, respectively. Slopes and intercepts from simple linear regression models of the two clotted erythrocyte fraction methods were not found to be significantly different (p > 0.05) when compared to the standard blood lead assay method. Modeled blood lead predicted from clotted erythrocyte fraction was evaluated at a test threshold of 3 µg/dL was found to have diagnostic sensitivity of 88% and specificity of 100%. Results from this study demonstrate clotted erythrocyte fraction samples are a viable alternative biological sample for retrospective public health surveillance of environmental exposure to lead.

Acute in vivo pulmonary toxicity assessment of occupationally relevant particulate matter from a cellulose nanofiber board.

Cellulose nanofibers (CNFs) are an emerging engineered nanomaterial that are utilized in a variety of applications, including as a replacement for urea-formaldehyde, and other adhesives, as the binding agent in manufactured fiber and particle boards. To ensure the health and well-being of those producing, installing, or otherwise using cellulose nanofiber boards (CNFBs) it is imperative that the particulate matter (PM) produced during CNFB manipulation be evaluated for toxicity. We developed and internally verified a generation system to examine the PM produced by sanding CNFB using aluminum oxide sandpaper. With 80-grit sandpaper our system produced a low dispersity aerosol, as determined by a scanning mobility particle sizer and an optical particle counter, with a geometric mean of 28 nm (GSD = 1.60). ICP-MS evaluation showed little difference in metal concentrations between CNFB PM and nonsanded CNFB stock. We then used the system to simultaneously generate and expose both male and female C57BL/6J mice acutely for 4 hours at a concentration of 7.9 mg/m3. Sham-exposed controls were treated similarly but without sanding the CNFB. Analysis of bronchoalveolar lavage (BAL) fluid biomarkers showed no signs of inflammatory response at either 4- or 24-hours post exposure. Further, BAL cell viability, number of total cells, and pulmonary cellular recruitment were not significantly changed between the sham-exposed controls and CNFB-exposed mice. Histology further confirmed no pulmonary toxicity as a result of CNFB PM inhalation. We conclude that inhalation of a high concentration of the PM from manipulation of a CNFB did not produce acute toxic responses within 24 hours of exposure.

Toxicity assessment of metal oxide nanomaterials using in vitro screening and murine acute inhalation studies.

Characterizations and in vitro toxicity screening were performed on metal oxide engineered nanomaterials (ENMs) independently comprising ZnO, CuO, CeO2, Fe2O3, WO3, V2O5, TiO2, Al2O3 and MgO. Nanomaterials that exhibited the highest toxicity responses in the in vitro screening assays (ZnO, CuO, and V2O5) and the lesser explored material WO3 were tested for acute pulmonary toxicity in vivo. Female and male mice (C57Bl/6J) were exposed to aerosolized metal oxide ENMs in a nose-only exposure system and toxicity outcomes (biomarkers of cytotoxicity, immunotoxicity, inflammation, and lung histopathology) at 4 and 24 h after the start of exposure were assessed. The studies were performed as part of the NIEHS Nanomaterials Health Implications Research consortium with the purpose of investigating the effects of ENMs on various biological systems. ENMs were supplied by the Engineered Nanomaterials Resource and Coordination Core. Among the ENMs studied, the highest toxicity was observed for CuO and ZnO NPs in both in vitro and in vivo acute models. Compared to sham-exposed controls, there was a significant increase in bronchoalveolar lavage neutrophils and proinflammatory cytokines and a loss of macrophage viability at both 4 h and 24 h for ZnO and CuO but not seen for V2O5 or WO3. These effects were observed in both female and male mice. The cell viability performed after in vitro exposure to ENMs and assessment of lung inflammation after acute inhalation exposure in vivo were shown to be sensitive endpoints to predict ENM acute toxicity.

Aquifer-Scale Observations of Iron Redox Transformations in Arsenic-Impacted Environments to Predict Future Contamination.

Iron oxides control the mobility of a host of contaminants in aquifer systems, and the microbial reduction of iron oxides in the subsurface is linked to high levels of arsenic in groundwater that affects greater than 150 million people globally. Paired observations of groundwater and solid-phase aquifer composition are critical to understand spatial and temporal trends in contamination and effectively manage changing water resources, yet field-representative mineralogical data are sparse across redox gradients relevant to arsenic contamination. We characterize iron mineralogy using X-ray absorption spectroscopy across a natural gradient of groundwater arsenic contamination in Vietnam. Hierarchical cluster analysis classifies sediments into meaningful groups delineating weathering and redox changes, diagnostic of depositional history, in this first direct characterization of redox transformations in the field. Notably, these groupings reveal a signature of iron minerals undergoing active reduction before the onset of arsenic contamination in groundwater. Pleistocene sediments undergoing postdepositional reduction may be more extensive than previously recognized due to previous misclassification. By upscaling to similar environments in South and Southeast Asia via multinomial logistic regression modeling, we show that active iron reduction, and therefore susceptibility to future arsenic contamination, is more widely distributed in presumably pristine aquifers than anticipated.

Skeletal toxicity resulting from exposure of growing male rats to coplanar PCB 126 is associated with disruption of calcium homeostasis and the GH-IGF-1 axis and direct effects on bone formation.

Skeletal toxicity has been reported following exposure to polychlorinated biphenyl (PCB) mixtures. However, molecular mechanisms remain poorly understood. We exposed groups of male 4-5-week-old Sprague-Dawley rats to 3,3', 4, 4', 5-pentachlorobiphenyl (PCB 126), a dioxin-like coplanar PCB congener by a single i.p. injection of 5 µmol/kg in soy oil vehicle or vehicle alone. After 4 weeks, rats were euthanized. PCB exposure resulted in hypocalcemia (P < 0.05) and significant increases in serum PTH without changes in serum phosphorous. Hyperparathyroidism was accompanied by increased expression of mRNAs of vitamin D3 metabolizing cytochrome P450 enzymes CYP27B1 and CYP24 in the kidney (P < 0.05). PCB exposure also reduced body weight, serum IGF-1, and hepatic expression of mRNAs encoding the male-specific GH-pattern-regulated CYP2C11 and CYP3A2 relative to controls (P < 0.05). PCB exposure reduced long bone length, diameter, and surface area, but increased trabecular thickness and volume (P < 0.05). Serum osteocalcin (P < 0.05), a marker and a regulator of bone formation, was reduced, but PCB exposure had no effect on the bone resorption marker RatLaps. Exposure of human intestinal Caco-2 cells to 10-100 nM PCB 126 in the presence of vitamin D3 resulted in inhibition of mRNAs for the calcium transporters TRPV6 and PMCA1b (P < 0.05). In addition, PCB 126 suppressed osteoblastogenesis in primary bone marrow mesenchymal stem cell cultures which was blunted by the AhR antagonist CH-223191. These data provide novel evidence that skeletal toxicity after exposure to PCB 126 is a result of disruption of calcium homeostasis and the GH-IGF-1 axis, and involves direct AhR-mediated effects on bone formation.

Correction: Worldwide increased prevalence of human adenovirus type 3 (HAdV-3) respiratory infections is well correlated with heterogeneous hypervariable regions (HVRs) of hexon.

[This corrects the article DOI: 10.1371/journal.pone.0194516.].

Worldwide increased prevalence of human adenovirus type 3 (HAdV-3) respiratory infections is well correlated with heterogeneous hypervariable regions (HVRs) of hexon.

Human adenovirus type 3 (HAdV-3) respiratory infections occurs worldwide in both children and adults, leading to severe morbidity and mortality, particularly in the paediatric age group and especially in neonates. During HAdV infection, neutralizing antibodies are formed against the epitopes located in the hyper variable regions (HVRs) of the hexon protein. These neutralizing antibodies provide protection against reinfection by viruses of the same type. Therefore it is reasonable to speculate that variations of HAdV-3 in the HVRs could impair the immunity acquired by previous infection with a different strain with variation in its HVRs. HAdV-3 has recently become the major agent of acute respiratory infection worldwide, being responsible for 15% to 87% of all adenoviral respiratory infections. However, despite the increased prevalence of HAdV-3 as respiratory pathogen, the diversity of hexon proteins in circulating strains remains unexplored. This study was designed to explore the variation in HVRs of hexon among globally distributed strains of HAdV-3 as well as to discover possible relationship among them, thus possibly shedding light on the cause for the increased prevalence of HAdV-3. In this study, for the first time we analysed the hexon proteins of all 248 available strains of HAdV-3 from the NCBI database and compared them with those of the HAdV-3 prototype (GB stain). We found that the HVRs of HAdV-3 strains circulating worldwide were highly heterogeneous and have been mutating continuously since -their original isolation. Based on their immense heterogeneity, the strains can be categorized into 25 hexon variants (3Hv-1 to 3Hv-25), 4 of which (3Hv-1 to 3Hv-4) comprises 80% of the strains. This heterogeneity may explain why HAdV-3 has become the most prevalent HAdVs type worldwide. The heterogeneity of hexon proteins also shows that the development of a vaccine against HAdV-3 might be challenging. The data on hexon variants provided here may be useful for the future epidemiological study of HAdV-3 infection.

Quantitative drinking water arsenic concentrations in field environments using mobile phone photometry of field kits.

Arsenic (As) groundwater contamination is common yet spatially heterogeneous within most environments. It is therefore necessary to measure As concentrations to determine whether a water source is safe to drink. Measurement of As in the field involves using a test strip that changes color in the presence of As. These tests are relatively inexpensive, but results are subjective and provide binned categorical data rather than exact determinations of As concentration. The goal of this work was to determine if photos of field kit test strips taken on mobile phone cameras could be used to extract more precise, continuous As concentrations. As concentrations for 376 wells sampled from Araihazar, Bangladesh were analyzed using ICP-MS, field kit and the new mobile phone photo method. Results from the field and lab indicate that normalized RGB color data extracted from images were able to accurately predict As concentrations as measured by ICP-MS, achieving detection limits of 9.2µg/L, and 21.9µg/L for the lab and field respectively. Data analysis is most consistent in the laboratory, but can successfully be carried out offline following image analysis, or on the mobile phone using basic image analysis software. The accuracy of the field method was limited by variability in image saturation, and variation in the illumination spectrum (lighting) and camera response. This work indicates that mobile phone cameras can be used as an analytical tool for quantitative measures of As and could change how water samples are analyzed in the field more widely, and that modest improvements in the consistency of photographic image collection and processing could yield measurements that are both accurate and precise.