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Over 40% of veterans from the Persian Gulf War (GW) (1990-1991) suffer from Gulf War Illness (GWI). Thirty years since the GW, the exposure and mechanism contributing to GWI remain unclear. One possible exposure that has been attributed to GWI are chemical warfare agents (CWAs). While there are treatments for isolated symptoms of GWI, the number of respiratory and cognitive/neurological issues continues to rise with minimum treatment options. This issue does not only affect veterans of the GW, importantly these chronic multisymptom illnesses (CMIs) are also growing amongst veterans who have served in the Afghanistan-Iraq war. What both wars have in common are their regions and inhaled exposures. In this review, we will describe the CWA exposures, such as sarin, cyclosarin, and mustard gas in both wars and discuss the various respiratory and neurocognitive issues experienced by veterans. We will bridge the respiratory and neurological symptoms experienced to the various potential mechanisms described for each CWA provided with the most up-to-date models and hypotheses.

Purpose Assess family-level factors associated with childhood immunization schedule adherence. Design Prospective cohort; Setting; The Healthy Start study enrolled 1,410 pregnant women in Denver, Colorado 2009-2014 Subjects Children with available vaccination data in medical records (0-6 years old) Measures Vaccine schedule completion and compliance Analysis Logistic regression comparing family-level factors that differ based on vaccine schedule adherence Results Most immunizations required in Colorado for school entry were below national completion goals with 61.8% of participants (n = 532/861) completing the full vaccination series. Most participants received the first dose of individual vaccines on time (73.5% - 90.7%), but fewer received all doses on time (21.0% - 39.5%). Factors associated with not completing the vaccination series (OR [95% CI]) included: in-utero exposure to cigarette smoke (1.97 [1.41, 2.75]), single parent household (1.70 [1.21, 2.38]), children identified as non-White (Hispanic 1.40 [1.01, 1.94]; Black 1.88 [1.24, 2.85]; Other 2.17 [1.34, 3.49]), mothers not working outside the home (1.98 [1.46, 2.67]), and household income Conclusions These findings may help identify groups at risk of immunization schedule non-adherence and may be used to target education/advocacy campaigns to reduce hesitancy and increase access in these populations.

Silica nanoparticles (SiNPs) released during the burning of sugarcane have been postulated to have a role in chronic kidney disease of unknown etiology. We tested the hypothesis that pristine SiNPs of the size present in sugarcane might cause chronic kidney injury when administered through the lung in rats. We administered 200- or 300-nm amorphous SiNPs twice weekly (4 mg/dose), or vehicle by oropharyngeal aspiration for 13 wk to rats followed by euthanasia after an additional 13 wk (26 wk total). Tissues were evaluated for the presence of SiNPs and evidence of histological injury. Both sizes of SiNPs caused kidney damage, with early tubular injury and inflammation (at week 13) that continued to inflammation and chronic fibrosis at week 26 despite discontinuation of the SiNP administration. Both sizes of SiNPs caused local inflammation in the lung and kidney and were detected in the serum and urine at week 13, and the 200-nm particles were also localized to the kidney with no evidence of retention of the 300-nm particles. At week 26, there was some clearance of the 200-nm silica from the kidneys, and urinary levels of SiNPs were reduced but still significant in both 200- and 300 nm-exposed rats. In conclusion, inhaled SiNPs cause chronic kidney injury that progresses despite stopping the SiNP administration. These findings support the hypothesis that human exposure to amorphous silica nanoparticles found in burned sugarcane fields could have a participatory role in chronic kidney disease of unknown etiology. NEW & NOTEWORTHY Inhalation of silica nanoparticles (SiNPs) released during the burning of sugarcane has been postulated to have a role in chronic kidney disease of unknown etiology (CKDu). We administered 200- and 300-nm amorphous SiNPs to rats by aspiration and observed kidney damage with tubular injury and inflammation that persisted even after stopping the SiNP exposure. These findings support the hypothesis that human exposure to SiNPs found in sugarcane ash could have a participatory role CKDu.

The devastating effects of the coronavirus disease 2019 (COVID-19) pandemic have made clear a global necessity for antiviral strategies. Most fatalities associated with infection from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) result at least partially from uncontrolled host immune response. Here, we use an antisense compound targeting a previously identified microRNA (miRNA) linked to severe cases of COVID-19. The compound binds specifically to the miRNA in question, miR-2392, which is produced by human cells in several disease states. The safety and biodistribution of this compound were tested in a mouse model via intranasal, intraperitoneal, and intravenous administration. The compound did not cause any toxic responses in mice based on measured parameters, including body weight, serum biomarkers for inflammation, and organ histopathology. No immunogenicity from the compound was observed with any administration route. Intranasal administration resulted in excellent and rapid biodistribution to the lungs, the main site of infection for SARS-CoV-2. Pharmacokinetic and biodistribution studies reveal delivery to different organs, including lungs, liver, kidneys, and spleen. The compound was largely cleared through the kidneys and excreted via the urine, with no accumulation observed in first-pass organs. The compound is concluded to be a safe potential antiviral treatment for COVID-19.

Engineered nanomaterials are becoming increasingly ubiquitous in our society, with numerous applications in medicine, consumer products, bioremediation, and advanced materials. As these nanomaterials increase in variety, analyzing their characteristics is of great importance. Single particle inductively coupled plasma-mass spectrometry (SP-ICP-MS) is a high-throughput, sensitive, and robust instrumental analysis method used to simultaneously characterize and quantify nanoparticles in a variety of matrices. One such type of nanoparticle of interest is amorphous silica nanoparticles (SiNPs). SiNPs have widespread use in consumer products such as food and cosmetics and are prime candidates for novel medical applications and uses in environmental bioremediation. Despite their increased use, SiNPs have been shown to have toxicological properties in vitro and in vivo, particularly with regard to the immune system. Because of the potential for increased SiNP exposure in the general public and in occupational settings, examining the relationship that SiNPs have with immune cells such as macrophages to elucidate mechanisms of toxicity is vital. To effectively determine the toxicity of nanoparticles, it is critical to examine dosimetry and the amount of nanoparticles taken up by the cell of interest. Different cell types have different uptake profiles, and varying physicochemical properties govern nanoparticle dosimetry and uptake in cells. Here, we describe a protocol using SP-ICP-MS to quantify and characterize the size, size distribution, and amount of SiNPs present in a cell and medium sample. We use a single-step digestion, which allows for the digestion of biological matrices while simultaneously keeping the SiNPs intact for SP-ICP-MS analysis. Clinically, this approach has the potential to be used as a method for analyzing SiNPs in other biological matrices, potentially as a way of defining SiNP uptake as a biomarker in immune-mediated diseases. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Treatment of cells with silica nanoparticles (SiNPs) and digestion of biological matrices Support Protocol 1: Culturing RAW 264.7 cells for SiNP uptake assay Support Protocol 2: Determination of SiNP size via dynamic light scattering Support Protocol 3: Optimization of sample and ICP-MS parameters for SP-ICP-MS analysis of cells and medium Basic Protocol 2: Analysis and quantification of SiNP uptake in macrophages with SP-ICP-MS.

Climate change should be of special concern for the nephrologist, as the kidney has a critical role in protecting the host from dehydration, but it is also a favorite target of heat stress and dehydration. Here we discuss how rising temperatures and extreme heat events may affect the kidney. The most severe presentation of heat stress is heat stroke, which can result in severe electrolyte disturbance and both acute and chronic kidney disease (CKD). However, lesser levels of heat stress also have multiple effects, including exacerbating kidney disease and precipitating cardiovascular events in subjects with established kidney disease. Heat stress can also increase the risk for kidney stones, cause multiple electrolyte abnormalities and induce both acute and chronic kidney disease. Recently there have been multiple epidemics of CKD of uncertain etiology in various regions of the world, including Mesoamerica, Sri Lanka, India and Thailand. There is increasing evidence that climate change and heat stress may play a contributory role in these conditions, although other causes, including toxins, could also be involved. As climate change worsens, the nephrologist should prepare for an increase in diseases associated with heat stress and dehydration.

Studies have suggested imatinib mesylate (ImM) as a potential treatment for systemic lupus erythematosus nephritis (SLEN). However, ImM has limited renal excretion. The goal of the current research was to develop an ImM containing nanoformulation, conduct studies to evaluate pharmacokinetics, and determine whether kidney deposition can be enhanced in a mouse model of SLEN. A fish oil–based ImM oil-in-water nanoemulsion was developed and characterized for particle size, zeta potential, pH, and stability. MRL/MpJ-Faslrp (model of SLEN) and MRL/MpJ (control) mice (12–13 weeks) received one dose of ImM as either a nanoemulsion or naked drug. Pharmacokinetics and kidney deposition studies were performed. Statistics were conducted with a student’s T-test. The nanoemulsion characteristics included particle size range of 60–80 nm, zeta potential of −6.6 to −7.8 mV, polydispersity index

As the world braces to enter its third year in the coronavirus disease 2019 (COVID-19) pandemic, the need for accessible and effective antiviral therapeutics continues to be felt globally. The recent surge of Omicron variant cases has demonstrated that vaccination and prevention alone cannot quell the spread of highly transmissible variants. A safe and nontoxic therapeutic with an adaptable design to respond to the emergence of new variants is critical for transitioning to treatment of COVID-19 as an endemic disease. Here, we present a novel compound, called SBCoV202, that specifically and tightly binds the translation initiation site of RNA-dependent RNA polymerase within the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome, inhibiting viral replication. SBCoV202 is a Nanoligomer,™ a molecule that includes peptide nucleic acid sequences capable of binding viral RNA with single-base-pair specificity to accurately target the viral genome. The compound has been shown to be safe and nontoxic in mice, with favorable biodistribution, and has shown efficacy against SARS-CoV-2 in vitro. Safety and biodistribution were assessed after three separate administration methods, namely intranasal, intravenous, and intraperitoneal. Safety studies showed the Nanoligomer caused no outward distress, immunogenicity, or organ tissue damage, measured through observation of behavior and body weight, serum levels of cytokines, and histopathology of fixed tissue, respectively. SBCoV202 was evenly biodistributed throughout the body, with most tissues measuring Nanoligomer concentrations well above the compound KD of 3.37 nM. In addition to favorable availability to organs such as the lungs, lymph nodes, liver, and spleen, the compound circulated through the blood and was rapidly cleared through the renal and urinary systems. The favorable biodistribution and lack of immunogenicity and toxicity set Nanoligomers apart from other antisense therapies, while the adaptability of the nucleic acid sequence of Nanoligomers provides a defense against future emergence of drug resistance, making these molecules an attractive potential treatment for COVID-19.

The increased use of amorphous silica nanoparticles (SiNPs) in food products, materials science, cosmetics, and pharmaceuticals has raised questions about potential hazards in the environment and in human health. Although SiNPs are generally thought to be benign, recent studies have demonstrated toxicity in different cell and animal models. Despite their ubiquitous use, SiNPs are rarely analyzed quantitatively. Often, the methods used to analyze silicon and SiNPs are difficult, costly, require the use of dangerous reagents, and are prone to interferences. Additionally, characterization of SiNPs in complex matrices requires extensive sample preparation. To address this, we propose a single-step digestion method for the determination of trace SiNP content in biological matrices. For conventional inductively coupled plasma-mass spectrometry (ICP-MS) analysis, biological samples are often digested with concentrated HNO

Single circulating factors are often investigated to explain air pollution-induced cardiovascular dysfunction, yet broader examinations of the identity and bioactivity of the entire circulating milieu remain understudied. The purpose of this study was to determine if exposure-induced cardiovascular dysfunction can be coupled with alterations in both serum bioactivity and the circulating proteome. Two cohorts of Spontaneously Hypertensive Rats (SHRs) were exposed to 150 or 500 μg/m3 diesel exhaust (DE) or filtered air (FA). In Cohort 1, we collected serum 1 hour after exposure for proteomics analysis and bioactivity measurements in rat aortic endothelial cells (RAECs). In Cohort 2, we assessed left ventricular pressure (LVP) during stimulation and recovery from the sympathomimetic dobutamine HCl, one day after exposure. Serum from DE-exposed rats had significant changes in 66 serum proteins and caused decreased NOS activity and increased VCAM-1 expression in RAECs. While rats exposed to DE demonstrated increased heart rate at the start of LVP assessments, heart rate, systolic pressure, and double product fell below baseline in DE-exposed rats compared to FA during recovery from dobutamine, indicating dysregulation of post-exertional cardiovascular function. Taken together, a complex and bioactive circulating milieu may underlie air pollution-induced cardiovascular dysfunction.

Sulfur mustard (SM) has been widely used as a chemical warfare agent including most recently in Syria. Mice exposed to SM exhibit an increase in pro-inflammatory cytokines followed by immune cell infiltration in the lung, however, the mechanisms leading to these inflammatory responses has not been completely elucidated. Mast cells are one of the first responding innate immune cells found at the mucosal surfaces of the lung and have been reported to be activated by SM in the skin. Therefore, we hypothesized that nitrogen mustard (NM: a surrogate for SM) exposure promotes activation of mast cells causing chronic respiratory inflammation. To assess the role of mast cells in NM-mediated pulmonary toxicity, we compared the effects of NM exposure between C57BL/6 and B6.Cg-KitW-sh/HNihrJaeBsmJ (KitW-sh; mast cell deficient) mice. Lung injury was observed in C57BL/6J mice following NM exposure (0.125 mg/kg) at 72 h, which was significantly abrogated in KitW-sh mice. Although both strains exhibited damage from NM, C57BL/6J mice had higher inflammatory cell infiltration and more elevated prostaglandin D2 (PGD2) present in bronchoalveolar lavage fluid compared with KitW-sh mice. Additionally, we utilized murine bone marrow-derived mast cells to assess NM-induced early and late activation. Although NM exposure did not result in mast cell degranulation, we observed an upregulation in PGD2 and IL-6 levels following exposure to NM. Results suggest that mast cells play a prominent role in lung injury induced by NM and may contribute to the acute and potentially long-term lung injury observed caused by SM.

Studies have suggested imatinib mesylate (ImM) as a potential treatment for systemic lupus erythematosus nephritis (SLEN). However, ImM has limited renal excretion. The goal of the current research was to develop an ImM containing nanoformulation, conduct studies to evaluate pharmacokinetics, and determine whether kidney deposition can be enhanced in a mouse model of SLEN. A fish oil-based ImM oil-in-water nanoemulsion was developed and characterized for particle size, zeta potential, pH, and stability. MRL/MpJ-Faslrp (model of SLEN) and MRL/MpJ (control) mice (12-13 weeks) received one dose of ImM as either a nanoemulsion or naked drug. Pharmacokinetics and kidney deposition studies were performed. Statistics were conducted with a student's T-test. The nanoemulsion characteristics included particle size range of 60-80 nm, zeta potential of -6.6 to -7.8 mV, polydispersity index 0.3, 3-day stability at 4 °C, and limited ImM leakage from the nanoemulsion in serum. Pharmacokinetics of the nanoformulation showed changes to pharmacokinetic parameters suggesting reduced systemic exposures (with reduced potential for toxicities) to ImM. Kidney deposition of ImM was threefold higher after 4 h in the MRL/MpJ-Faslrp mice that received the nanoformulation vs. naked drug. The current study showed encouraging results for development of a stable and well-characterized nanoemulsion for optimizing kidney deposition of ImM. Future strategies will define dose-efficacy and dose-toxicity relationships and evaluate approaches to further enhance kidney delivery and optimize deposition to the mesangial location of the kidney.

Eosinophils are a myeloid cell subpopulation that mediates type 2 T helper cell immune responses. Unexpectedly, we identified a rapid accumulation of eosinophils in 22 human liver grafts after hepatic transplantation. In contrast, no eosinophils were detectable in healthy liver tissues before transplantation. Studies with two genetic mouse models of eosinophil deficiency and a mouse model of antibody-mediated eosinophil depletion revealed exacerbated liver injury after hepatic ischemia and reperfusion. Adoptive transfer of bone marrow-derived eosinophils normalized liver injury of eosinophil-deficient mice and reduced hepatic ischemia and reperfusion injury in wild-type mice. Mechanistic studies combining genetic and adoptive transfer approaches identified a critical role of suppression of tumorigenicity (ST2)-dependent production of interleukin-13 by eosinophils in the hepatoprotection against ischemia-reperfusion-induced injury. Together, these data provide insight into a mechanism of eosinophil-mediated liver protection that could serve as a therapeutic target to improve outcomes of patients undergoing liver transplantation.

Mast cells are important effector cells in the immune system and undergo activation (i.e., degranulation) by two major mechanisms: IgE-mediated and non-IgE–mediated mechanisms. Although IgE-mediated degranulation is well researched, the cellular mechanisms of non-IgE–mediated mast cell activation are poorly understood despite the potential to induce similar pathophysiological effects. To better understand non-IgE mast cell degranulation, we characterized and compared cellular metabolic shifts across several mechanisms of degranulation (allergen-induced [IgE-mediated], 20 nm of silver nanoparticle-mediated [non-IgE], and compound 48/80-mediated [non-IgE]) in murine bone marrow–derived mast cells. All treatments differentially impacted mitochondrial activity and glucose uptake, suggesting diverging metabolic pathways between IgE- and non-IgE–mediated degranulation. Non-IgE treatments depleted mast cells’ glycolytic reserve, and compound 48/80 further inhibited the ability to maximize mitochondrial respiration. This cellular reprogramming may be indicative of a stress response with non-IgE treatments. Neither of these outcomes occurred with IgE-mediated degranulation, hinting at a separate programmed response. Fuel flexibility between the three primary mitochondrial nutrient sources was also eliminated in activated cells and this was most significant in non-IgE–mediated degranulation. Lastly, metabolomics analysis of bone marrow–derived mast cells following degranulation was used to compare general metabolite profiles related to energetic pathways. IgE-mediated degranulation upregulated metabolite concentrations for the TCA cycle and glycolysis compared with other treatments. In conclusion, mast cell metabolism varies significantly between IgE- and non-IgE–mediated degranulation suggesting novel cell regulatory mechanisms are potentially driving unexplored pathways of mast cell degranulation.

Engineered nanomaterials (ENMs) are being incorporated at an unprecedented rate into consumer and biomedical products. This increased usage will ultimately lead to increased human exposure; therefore, understanding ENM safety is an important concern to the public. Although ENMs may exert toxicity through multiple mechanisms, one common mechanism of toxicity recognized across a range of ENMs with varying physicochemical properties is oxidative stress. Further, it is recognized that several key physicochemical properties of ENMs including size, material composition, surface chemistry, band gap, and level of ionic dissolution for example contribute to ENM driven oxidative stress. While it has been shown that exposure of cells to ENMs at high acute doses produce reactive oxygen species at a toxic level often leading to cytotoxicity, there is little research looking at oxidative stress caused by ENM exposure at more relevant low or non-toxic doses. Although the former can lead to apoptosis, genotoxicity, and inflammation, the latter can potentially be damaging as chronic changes to the intracellular redox state leads to cellular reprogramming, resulting in disease initiation and progression among other systemic damage. This current opinions article will review the physicochemical properties and mechanisms associated with ENM-driven oxidative stress and will discuss the need for research investigating effects on the redox proteome that may lead to cellular dysfunction at low or chronic doses of ENMs.

The broad list of commercial applications for multi-walled carbon nanotubes (MWCNT) can be further expanded with the addition of various surface chemistry modifications. For example, standard commercial grade MWCNT (C-grade) can be carboxylated (COOH) or nitrogen-doped (N-doped) to suite specific utilities. We previously reported dose-dependent expansions of cardiac ischemia/reperfusion (I/R) injury, 24 hours after intratracheal instillation of C-grade, COOH, or N-doped MWCNT in mice. Here we have tested the hypothesis that airway exposure to MWCNT perturbs cardiovascular adenosinergic signaling, which could contribute to exacerbation of cardiac I/R injury. One hundred microliters of Vehicle or identical suspension volumes containing 100 μg of C-grade, COOH, or N-doped MWCNT were instilled into the trachea of CD-1 ICR mice. One day later we measured cyclic adenosine monophosphate (cAMP) concentrations in cardiac tissue and evaluated arterial adenosinergic smooth muscle signaling mechanisms related to nitric oxide synthase (NOS) and cyclooxygenase (COX) in isolated aortic tissue. We also verified cardiac I/R injury expansion and examined both lung histology and bronchoalveolar lavage fluid cellularity in MWCNT exposed mice. Myocardial cAMP concentrations were reduced (p < 0.05) in the C-grade group by 17.4% and N-doped group by 13.7% compared to the Vehicle group. Curve fits to aortic ring 2-Cl-Adenosine concentration responses were significantly greater in the MWCNT groups vs. the Vehicle group. Aortic constrictor responses were more pronounced with NOS inhibition and were abolished with COX inhibition. These findings indicate that addition of functional chemical moieties on the surface of MWCNT may alter the biological responses to exposure by influencing cardiovascular adenosinergic signaling and promoting cardiac injury.

Achyut J Raghavendra,1 Wren E Gregory,1 Tyler J Slonecki,2 Yongchang Dong,1 Indushekhar Persaud,3 Jared M Brown,3 Terri F Bruce,2 Ramakrishna Podila1,4 1Laboratory of Nano-Biophysics, Department of Physics and Astronomy, Clemson Nanomaterials Institute, Clemson University, Clemson, SC, USA; 2Clemson Light Imaging Facility, Clemson University, Clemson, SC, USA; 3Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado-Anschutz Medical Campus, Aurora, CO, USA; 4Clemson University School of Health Research and COMSET, Clemson University, Clemson, SC, USA Background: Although optical spectroscopy promises improved lateral resolution for cancer imaging, its clinical use is seriously impeded by background fluorescence and photon attenuation even in the so-called two-photon absorption (2PA) imaging modality. An efficient strategy to meet the clinical cancer imaging needs, beyond what two-photon absorption (2PA) offers, is to use longer excitation wavelengths through three-photon absorption (3PA). A variety of fluorescent dyes and nanoparticles (NPs) have been used in 3PA imaging. However, their non-linear 3PA coefficient is often low necessitating high excitation powers, which cause overheating, photodamage, and photo-induced toxicity. Doped wide band gap semiconductors such as Mn:ZnS NPs have previously been used for 3PA but suffer from poor 3PA coefficients. Methods: Here, we prepared ZnO NPs with intrinsic defects with high 3PA coefficients using a polyol method. We functionalized them with peptides for selective uptake by glioblastoma U87MG cells and used breast cancer MCF-7 cells as control for 3PA studies. Uptake was measured using inductively coupled plasma-mass spectrometry. Biocompatibility studies were performed using reactive oxygen species and cell viability assays. Results: We demonstrate that ZnO NPs, which have a band gap of 3.37 eV with an order of magnitude higher 3PA coefficients, can facilitate the use of longer excitation wavelengths 950–1,100 nm for bioimaging. We used the presence intrinsic defects (such as O interstitials and Zn vacancies) in ZnO NPs to induce electronic states within the band gap that can support strong visible luminescence 550–620 nm without the need for extrinsic doping. The peptide functionalization of ZnO NPs showed selective uptake by U87MG cells unlike MCF-7 cells without the integrin receptors. Furthermore, all ZnO NPs were found to be biocompatible for 3PA imaging. Conclusion: We show that defect-induced luminescence 550–620 nm in ZnO NPs (20 nm) due to 3PA at longer excitation (975 nm) can be used for 3PA imaging of U87MG glioblastoma cells with lower background noise. Keywords: three-photon imaging, ZnO nanoparticles, defects, photoluminescence

Background & Aim Alcohol consumption increases intestinal permeability and causes damage to hepatocytes, leading to the release of pathogen- and damage-associated molecular pattern molecules (PAMPs and DAMPs), stimulating hepatic macrophages and activating NF-κB. The resultant inflammation exacerbates alcoholic liver disease (ALD). However, much less is known about the mechanisms attenuating inflammation and preventing disease progression in most heavy drinkers. Interleukin (IL)-33 is a DAMP (alarmin) released from dead cells that acts through its receptor, IL-1 receptor like 1 (ST2). ST2 signaling has been reported to either stimulate or inhibit NF-κB activation. The role of IL-33/ST2 in ALD has not been studied. Methods Serum levels of IL-33 and its decoy receptor, soluble ST2 (sST2) were measured in ALD patients. Alcohol-induced liver injury, inflammation and hepatic macrophage activation were compared between wild-type, IL-33 −/− and ST2 −/− mice in several models. Results Elevation of serum IL-33 and sST2 were only observed in patients with severe decompensated ALD. Consistently, in mice with mild ALD without significant cell death and IL-33 release, IL-33 deletion did not affect alcohol-induced liver damage. However, ST2-deletion exacerbated ALD, through enhancing NF-κB activation in liver macrophages. In contrast, when extracellular IL-33 was markedly elevated, liver injury and inflammation were attenuated in both IL-33 −/− and ST2 −/− mice compared to wild-type mice. Conclusion Our data revealed a dichotomous role of IL-33/ST2 signaling during ALD development. At early and mild stages, ST2 restrains the inflammatory activation of hepatic macrophages, through inhibiting NF-κB, and plays a protective function in an IL-33-independent fashion. During severe liver injury, significant cell death and marked IL-33 release occur, which triggers IL-33/ST2 signaling and exacerbates tissue damage. Lay summary In mild ALD, ST2 negatively regulates the inflammatory activation of hepatic macrophages, thereby protecting against alcohol-induced liver damage, whereas in the case of severe liver injury, the release of extracellular IL-33 may exacerbate tissue inflammation by triggering the canonical IL-33/ST2L signaling in hepatic macrophages.

Ball-milling utilizes mechanical stress to modify properties of carbon nanotubes (CNTs) including size, capping, and functionalization. Ball-milling, however, may introduce structural defects resulting in altered CNT-biomolecule interactions. Nanomaterial-biomolecule interactions result in the formation of the biocorona (BC), which alters nanomaterial properties, function, and biological responses. The formation of the BC is governed by the nanomaterial physicochemical properties and the physiological environment. Underlying disease states such as cardiovascular disease can alter the biological milieu possibly leading to unique BC identities. In this ex vivo study, we evaluated variations in the formation of the BC on single-walled CNTs (SWCNTs) due to physicochemical alterations in structure resulting from ball-milling and variations in the environment due to the high-cholesterol disease state. Increased ball-milling time of SWCNTs resulted in enhanced structural defects. Following incubation in normal mouse serum, label-free quantitative proteomics identified differences in the biomolecular content of the BC due to the ball-milling process. Further, incubation in cholesterol-rich mouse serum resulted in the formation of unique BCs compared to SWCNTs incubated in normal serum. Our study demonstrates that the BC is modified due to physicochemical modifications such as defects induced by ball-milling and physiological disease conditions, which may result in variable biological responses.

Mast cells are an essential cell type within the immune system, playing a prominent role in a variety of cell functions—both adaptive and innate immune system as well as others. This central role throughout many systems means that any inappropriate modulation of mast cell activation can, and does, contribute to many pathologies, including lung disease, autoimmune disorders, and cancer as examples. There are many exogenous compounds that can lead to activation of mast cells and potentially cause irregular function, one of which are engineered nanomaterials (ENMs). The human population is ubiquitously exposed to a range of ENM at a high rate through exposure mechanisms associated with high mast cell number, making these nanoparticles suspect when considering mast cell dysfunction. Research looking into ENM-induced mast cell pathologies is underwhelming; however; current research on the topic shows a clear ability for ENM to change mast cell activity and must be further explored.

The mucosal membranes (i.e. oral, and nasal cavities, gut and female reproductive tract) throughout the body are ideal locations for drug delivery as they result in rapid uptake of the pharmaceutical into the blood system; however, these membranes are protected by the mucosal immune system (MIS). This intricate network of epithelial cells, microbiota, immune cells and mucus present a unique challenge for drug delivery due to their barrier functions and responses to foreign pathogens. There is a constant need to improve drug delivery methods, particularly in regard to mucosal drug delivery. The application of engineered nanomaterials (ENM), due to their unique physicochemical properties, have significant potential to improve mucosal drug delivery. This chapter will discuss key physicochemical properties (i.e. size, shape, surface functionalization, and solubility) and how modification of these properties can alter the biological impact of ENMs. Finally, a brief overview of the normal function of key MIS cells (i.e. epithelial cells, goblet cells, Paneth cells, dendritic cells, macrophages, T and B lymphocytes, mast cells, and endothelial cells) and how ENM exposure alters their function will be presented.

Coal is one of the most abundant and economic sources for global energy production. However, the burning of coal is widely recognized as a significant contributor to atmospheric particulate matter linked to deleterious respiratory impacts. Recently, we have discovered that burning coal generates large quantities of otherwise rare Magnéli phase titanium suboxides from TiO2 minerals naturally present in coal. These nanoscale Magnéli phases are biologically active without photostimulation and toxic to airway epithelial cells in vitro and to zebrafish in vivo. Here, we sought to determine the clinical and physiological impact of pulmonary exposure to Magnéli phases using mice as mammalian model organisms. Mice were exposed to the most frequently found Magnéli phases, Ti6O11, at 100 parts per million (ppm) via intratracheal administration. Local and systemic titanium concentrations, lung pathology, and changes in airway mechanics were assessed. Additional mechanistic studies were conducted with primary bone marrow derived macrophages. Our results indicate that macrophages are the cell type most impacted by exposure to these nanoscale particles. Following phagocytosis, macrophages fail to properly eliminate Magnéli phases, resulting in increased oxidative stress, mitochondrial dysfunction, and ultimately apoptosis. In the lungs, these nanoparticles become concentrated in macrophages, resulting in a feedback loop of reactive oxygen species production, cell death, and the initiation of gene expression profiles consistent with lung injury within 6 weeks of exposure. Chronic exposure and accumulation of Magnéli phases ultimately results in significantly reduced lung function impacting airway resistance, compliance, and elastance. Together, these studies demonstrate that Magnéli phases are toxic in the mammalian airway and are likely a significant nanoscale environmental pollutant, especially in geographic regions where coal combustion is a major contributor to atmospheric particulate matter. This work was financially supported by the Virginia-Maryland College of Veterinary Medicine, the National Natural Science Foundation of China (41522111 and 4176114462), The Center for the Environmental Implications of Nanotechnology (NSF Cooperative Agreement EF-0830093), the Virginia Tech Institute for Critical Technology and Applied Science (ICTAS), the Open Foundation of East China Normal University, the Virginia Tech National Center for Earth and Environmental Nanotechnology Infrastructure (NSF Cooperative Agreement ECCS-1542100), the STAR Fellowship Assistance Agreement no. FP-91780101-1 awarded by the U.S. Environmental Protection Agency (EPA), The National Institutes of Health (R01 ES019311). Student work on this publication was supported by the National Institute of Allergy and Infectious Diseases Animal Model Research for Veterinarians (AMRV) training grant (T32-OD010430) and the American Association of Immunologist Careers in Immunology Fellowship Program (VR-S).

Zinc oxide nanoparticles (ZnO NPs) are used in numerous applications, including sunscreens, cosmetics, textiles, and electrical devices. Increased consumer and occupational exposure to ZnO NPs potentially poses a risk for toxicity. While many studies have examined the toxicity of ZnO NPs, little is known regarding the toxicological impact of inherent defects arising from batch-to-batch variations. It was hypothesized that the presence of varying chemical defects in ZnO NPs will contribute to cellular toxicity in rat aortic endothelial cells (RAECs). Pristine and defected ZnO NPs (oxidized, reduced, and annealed) were prepared and assessed three major cellular outcomes; cytotoxicity/apoptosis, reactive oxygen species production and oxidative stress, and endoplasmic reticulum (ER) stress. ZnO NPs chemical defects were confirmed by X-ray photoelectron spectroscopy and photoluminescence. Increased toxicity was observed in defected ZnO NPs compared to the pristine NPs as measured by cell viability, ER stress, and glutathione redox potential. It was determined that ZnO NPs induced ER stress through the PERK pathway. Taken together, these results demonstrate a previously unrecognized contribution of chemical defects to the toxicity of ZnO NPs, which should be considered in the risk assessment of engineered nanomaterials., by Indushekhar Persaud, Achyut J. Raghavendra, Archini Paruthi, Nasser B. Alsaleh, Valerie C. Minarchick, James R. Roede, Ramakrishna Podila and Jared M. Brown

Mast cells are a key effector cell in type I allergic reactions. It has been shown that environmental exposures such as diesel exhaust and heavy metals exacerbate mast cell degranulation and activation. Today, the use of engineered nanomaterials (ENMs) is rapidly expanding and silver nanoparticles (AgNP) are one of the mostly widely utilized ENMs, primarily for their antimicrobial properties, and are being incorporated into many consumer and biomedical products. We assessed whether pre-exposure of bone marrow-derived mast cells (BMMCs) to 20 nm AgNPs enhanced degranulation and activation to an allergen (dinitrophenol-conjugated human serum albumin) by measuring β-hexosaminidase release, LTB4 and IL-6 production. In addition, we assessed reactive oxygen species (ROS) generation, cell oxidative stress and toxicity as well as total and individual protein tyrosine phosphorylation (p-Tyr). We found that pre-exposure of BMMCs to AgNPs results in exacerbated allergen-mediated mast cell degranulation, LTB4 production and IL-6 release. Exposure of BMMCs to AgNPs exacerbated allergen-induced ROS generation, however, this was not associated with oxidative stress nor cell death. Finally, pre-exposure to AgNPs enhanced allergen-mediated global p-Tyr as well as individual proteins including Syk, PLCγ and LAT. Our findings indicate that pre-exposure to AgNPs exacerbates mast cell allergen-mediated phosphorylation of FceR1-linked tyrosine kinases and ROS generation resulting in amplified early and late-phase responses. These findings suggest that exposure to AgNPs has the potential to prime mast cells to allergic immune responses, which could be of particular concern to atopic populations as the use of AgNPs in consumer and biomedical products rapidly increases.

Nanoparticles under a reactive microenvironment, have the propensity to undergo morphological and compositional changes, which can translate into band edge widening. Although cell membrane depolarization has been linked with the electronic band structure of nanomaterials in their native state, the change in band structure as a consequence of a soluble nanoparticle system is less studied. Therefore we studied the consequence of dissolution of CuO nanoparticles on the band structure and flat band potentials and correlated it with its ability to induce a intracellular oxidative stress. The temporal variation in bandgap, fermi energy level and valence band maxima were evaluated on the remnant CuO nanoparticles post dissolution. CuO nanoparticles showed a very high dissolution in simulated body fluid (51%) and cell culture media (75%). This dissolution resulted in an in situ physico-chemical transformation of CuO nanoparticles. A temporal increase in the bandgap energy as a result of media interaction was up to 107%. Temporal variation in the flat band potentials with the generation of intracellular ROS, cell viability, late and early apoptosis in addition to necrosis on RAW 264.7 cells was established due to biological redox potential overlap. The mRNA expression for TNF-α, IL-6, IL-1β and IL-10 in response to the particle treatment was also evalulated for 6 hours. Through this study, we establish that the toxicological potential of CuO nanoparticles is a temporal function of band energies (its overlap with the intracellular redox potential) followed by release of ionic species in the cytotoxic regime.

Carbon nanotubes (CNTs) are of great interest for the development of drugs and vaccines due to their unique physicochemical properties. The high surface area to volume ratio and delocalized pi-electron cloud of CNTs promote binding of proteins to the surface forming a protein corona. This unique feature of CNTs has been recognized for potential delivery of antigens for strong and long-lasting antigen-specific immune responses. Based on an earlier study that demonstrated increased protein binding, we propose that carboxylated multiwalled CNTs (MWCNTs) can function as an improved carrier to deliver antigens such as ovalbumin (OVA). To test this hypothesis, we coated carboxylated MWCNTs with OVA and measured uptake and activation of antigen-presenting cells (macrophages) and their ability to stimulate CD4(+) T-cell proliferation. We employed two types of carboxylated MWCNTs with different surface areas and defects (MWCNT-2 and MWCNT-30). MWCNT-2 and MWCNT-30 have surface areas of ~215 m(2)/g and 94 m(2)/g, respectively. The ratios of D- to G-band areas (I D/I G) were 0.97 and 1.37 for MWCNT-2 and MWCNT-30, respectively, samples showing that MWCNT-30 contained more defects. The increase in defects in MWCNT-30 led to increased binding of OVA as compared to MWCNT-2 (1,066±182 μg/mL vs 582±41 μg/mL, respectively). Both types of MWCNTs, along with MWCNT-OVA complexes, showed no observable toxicity to bone-marrow-derived macrophages up to 5 days. Surprisingly, we found that MWCNT-OVA complex significantly increased the expression of major histocompatibility complex class II on macrophages and production of pro-inflammatory cytokines (tumor necrosis factor-α and interleukin 6), while MWCNTs without OVA protein corona did not. The coculture of MWCNT-OVA-complex-treated macrophages and OVA-specific CD4(+) T-cells isolated from OT-II mice demonstrated robust proliferation of CD4(+) T-cells. This study provides strong evidence for a role for defects in carboxylated MWCNTs and their use in the efficient delivery of antigens for the development of next-generation vaccines.

Nanoparticle (NP) association with macromolecules in a physiological environment forms a biocorona (BC), which alters NP distribution, activity, and toxicity. While BC formation is dependent on NP physicochemical properties, little information exists on the influence of the physiological environment. Obese individuals and those with cardiovascular disease exist with altered serum chemistry, which is expected to influence BC formation and NP toxicity. We hypothesize that a BC formed on NPs following incubation in hyperlipidemic serum will result in altered NP–BC protein content, cellular association, and toxicity compared to normal serum conditions. We utilized Fe3O4 NPs, which are being developed as MRI contrast and tumor targeting agents to test our hypothesis. We used rat aortic endothelial cells (RAECs) within a dynamic flow in vitro exposure system to more accurately depict the in vivo environment. A BC was formed on 20nm PVP-suspended Fe3O4 NPs following incubation in water, 10% normal or hyperlipidemic rat serum. Addition of BCs resulted in increased hydrodynamic size and decreased surface charge. More cholesterol associated with Fe3O4 NPs after incubation in hyperlipidemic as compared with normal serum. Using quantitative proteomics, we identified unique differences in BC protein components between the 2 serum types. Under flow conditions, formation of a BC from both serum types reduced RAECs association of Fe3O4 NPs. Addition of BCs was found to exacerbate RAECs inflammatory gene responses to Fe3O4 NPs (Fe3O4-hyperlipidemic > Fe3O4-normal > Fe3O4) including increased expression of IL-6, TNF-α, Cxcl-2, VCAM-1, and ICAM-1. Overall, these findings demonstrate that disease-induced variations in physiological environments have a significant impact NP-BC formation, cellular association, and cell response.

Chemical warfare agents (CWA) were widely used during the Gulf War (1990–1991) and as a result left ~30% of veterans suffering from Gulf War Illness (GWI). The vesicating agent, sulfur mustard (SM) is one of the most noteworthy CWA used during this time. Prior research has demonstrated that SM targets the bone marrow and has the potential to influence immune cells. In the skin, SM is reported to induce mast cell degranulation. Therefore, the aim of this study was to determine if nitrogen mustard (NM) (a surrogate for SM) promotes activation of mast cells. Using murine bone marrow derived mast cells (BMMCs) we assessed activation through degranulation, eicosanoid expression, and pro-inflammatory cytokine production. While NM exposure (1μM–50μM) did not result in mast cell degranulation, we observed a 30-fold increase of cyclooxygenase-2 (COX-2) mRNA production at 10μM at 6h. In addition, IL-6 production by BMMCs post 24h exposure to NM demonstrated a dose dependent increase indicative of late phase activation. To further demonstrate a pivotal role for mast cells in NM exposure, we compared the effects of NM exposure on lung pathology between C57BL/6 and B6.Cg-KitW-sh/ HNihrJaeBsmJ (mast cell deficient) mice. Significant lung injury was observed in C57BL/6J following NM exposure at 72 hrs as indicated by H&E staining. In contrast, significantly less injury was observed in mast cell deficient mice. Similarly, increases in total protein and IL-6 was observed in C57BL/6 mice but not mast cell deficient mice. These results suggest that mast cells play a prominent role in the injury induced by NM and may contribute to symptoms observed in GWI.

Non-IgE mediated pathways of mast cell degranulation result in similar pathological outcomes as IgE mediated degranulation without prior IgE sensitization thereby making it difficult to identify causative agents and develop therapeutics for patients. We have demonstrated that 20 nm silver nanoparticles (AgNP) induce non-IgE mast cell degranulation through an unknown mechanism. However, we have established that this non-IgE response is strain dependent with C57BL/6 bone marrow derived mast cells (BMMCs) being high responders and LP/J BMMCs being low responders. Further, RNA sequencing of high-responder and low-responder BMMCs demonstrated a significant increase in thioredoxin-interacting protein (txnip) in the low-responders after AgNP mediated degranulation. We explored this protein to determine its regulatory potential in non-IgE mast cell degranulation. At 1hr following exposure to AgNP, txnip mRNA levels increased in LP/J but not C57BL/6 which confirmed RNAseq results. Txnip protein levels were similarly decreased in BMMCs from both strains at 1hr; however, there was an increase in the low-responder BMMCs at six hours post-exposure that did not occur in the high-responder BMMCs. siRNA knockdown of txnip resulted in increased BMMC degranulation by AgNP. Txnip is strongly involved in cellular metabolism. Seahorse XfP results demonstrated a lower energy and glycolytically focused phenotype in low-responders compared to high-responders. C57BL/6 have significantly lowered glycolytic capacity after AgNP-mediated degranulation not present in LP/J. Our data suggests that txnip plays a role in non-IgE mediated mast cell degranulation and may modulate cellular metabolism.

Engineered nanomaterials (ENM) are being used in a wide range of consumer products and pharmaceuticals; hence, there is an increasing risk for human exposure and potential adverse outcomes. The immune system, vital in host defense and protection against environmental agents, is typically initiated and executed by innate effector immune cells including macrophages and neutrophils. Previous literature has reported the immune system as a major target of ENM toxicity; however, there is inconsistency regarding the immunotoxicity of ENM. This could be attributed to differences in ENM physicochemical properties, cellular models examined, biocorona formation, etc. Thus, the current study examined the toxicity and immunomodulatory effects of silver nanoparticles (AgNP), one of the most utilized ENM in consumer and medical products, in two key innate immune cell models, e.g. RAW 264.7 cells (macrophages) and differentiated MPRO 2.1 cells (promyelocytes/neutrophils). The results showed that despite a generation of reactive oxygen species, exposure to 20 nm citrate-coated AgNP was not associated with major oxidative damage, inflammatory responses, nor cytotoxicity. Nevertheless, and most importantly, pre-exposure to the AgNP for 24 h enhanced RAW 264.7 cell phagocytic ability as well as the release of inflammatory cytokine interleukin-6 in response to lipopolysaccharide (LPS). In MPRO 2.1 cells, AgNP pre-exposure also resulted in enhanced phagocytic ability; however, these cells manifest reduced cell degranulation (elastase release) and oxidative burst in response to phorbol myristate acetate (PMA). Taken together, these findings indicated to us that exposure to AgNP, despite not being directly (cyto)toxic to these cells, had the potential to alter immune cell responses. The findings underscore the import of assessing immune cell function post-exposure to ENM beyond the standard endpoints such as oxidative stress and cytotoxicity. In addition, these findings further illustrate the importance of understanding the underlying molecular mechanisms of ENM-cellular interactions, particularly in the immune system.

We carried out comprehensive structural, magnetic and toxicity studies of Mg1−xZnxFe2O4 and Cu1−xZnxFe2O4 ferrites that we plan to use as contrast agents for magnetic resonance-imaging thermometry. Samples were prepared by a standard ceramic technique. Their structural properties were investigated with an x-ray diffraction system, and data was analyzed using the Rietveld method that allowed us to determine cations distribution. Magnetic properties of MgZn and CuZn ferrites were studied using a superconducting quantum interference device magnetometer, a vibrating sample magnetometer, and Mossbauer spectrometer. Magnetometry measurements were cross-correlated with XRD results and compared with magnetic properties determined by Mossbauer spectroscopy, allowing us to understand the magnetic structure of studied ferrites. The temperature dependent magnetic measurements were used as guidance to select the most suitable MgZn and CuZn ferrites compositions for MRI thermometry near human body temperature. The inflammation studies indicated that MgZn ferrites are more suitable from the toxicology point of view for MRI thermometry.

Mast cells are key effector cells in inflammatory and allergic immune responses such as asthma, rhinitis, and atopic dermatitis. Activation of mast cells leads to immediate release of preformed mediators such as histamine and proteases, which can regulate vascular permeability and the function of a number of immune and nonimmune cells. Engineered nanomaterials (ENM) have been utilized for a wide range of applications and introduced into a number of consumer products; yet the consequent increase in human exposure and any potential adverse effects have not been fully evaluated. Modulation of the immune system function has been shown to be a major toxicological consequence of ENM exposure. The implication of mast cells in ENM-mediated toxicity, including the most widely utilized carbon and metal-based ENMs, has been previously demonstrated; and therefore, understanding direct ENM interaction with mast cells at the cellular and molecular level is of critical importance for the safe implementation of ENMs into consumer products.

Mast cells are key effector cells in inflammatory and allergic immune responses such as asthma, rhinitis, and atopic dermatitis. Activation of mast cells leads to immediate release of preformed mediators such as histamine and proteases, which can regulate vascular permeability and the function of a number of immune and nonimmune cells. Engineered nanomaterials (ENM) have been utilized for a wide range of applications and introduced into a number of consumer products; yet the consequent increase in human exposure and any potential adverse effects have not been fully evaluated. Modulation of the immune system function has been shown to be a major toxicological consequence of ENM exposure. The implication of mast cells in ENM-mediated toxicity, including the most widely utilized carbon and metal-based ENMs, has been previously demonstrated; and therefore, understanding direct ENM interaction with mast cells at the cellular and molecular level is of critical importance for the safe implementation of ENMs into consumer products.

Prior research has demonstrated cells exposed to silver nanoparticles (AgNPs) undergo endoplasmic reticulum (ER) stress leading to cellular apoptosis and toxicity, however, the fundamental mechanism underlying AgNP-induced ER stress is unknown. We hypothesize the biophysical interactions between AgNPs and adsorbed proteins lead to misfolded proteins to elicit an ER stress response. Our investigation examined rat aortic endothelial cells (RAEC) exposed to 20 or 100 nm AgNPs with or without a biocorona (BC) consisting of bovine serum albumin (BSA), high density lipoprotein (HDL) or fetal bovine serum (FBS) to form a complex BC. The presence of a BC consisting of BSA or FBS proteins significantly reduced uptake of 20 nm and 100 nm AgNPs in RAEC. Western blot analysis indicated robust activation of the IREα and PERK pathways in RAEC exposed to 20 nm despite the reduction in uptake by the presence of a BC. This was not observed for the 100 nm AgNPs. Hyperspectral darkfield microscopy qualitatively confirmed that the preformed BC was maintained following uptake by RAEC. Transmission electron microscopy demonstrated a size dependent effect on the sub-cellular localization of AgNPs. Overall, these results suggest that AgNP size, surface area and BC formation governs the induction of ER stress and alterations in intracellular trafficking.

Pulmonary instillation of multiwalled carbon nanotubes (MWCNT) has the potential to promote cardiovascular derangements, but the mechanisms responsible are currently unclear. We hypothesized that exposure to MWCNT would result in increased epithelial barrier permeability by 24 h postexposure and initiate a signaling process involving IL-6/gp130 transsignaling in peripheral vascular tissue. To test this hypothesis we assessed the impact of 1 and 10 μg/cm2MWCNT on transepithelial electrical resistance (TEER) and expression of barrier proteins and cell activation in vitro using normal human bronchial epithelial primary cells. Parallel studies using male Sprague-Dawley rats instilled with 100 μg MWCNT measured bronchoalveolar lavage (BAL) differential cell counts, BAL fluid total protein, and lung water-to-tissue weight ratios 24 h postexposure and quantified serum concentrations of IL-6, soluble IL-6r, and soluble gp130. Aortic sections were examined immunohistochemically for gp130 expression, and gp130 mRNA/protein expression was evaluated in rat lung, heart, and aortic tissue homogenates. Our in vitro findings indicate that 10 μg/cm2MWCNT decreased the development of TEER and zonula occludens-1 expression relative to the vehicle. In rats MWCNT instillation increased BAL protein, lung water, and induced pulmonary eosinophilia. Serum concentrations of soluble gp130 decreased, aortic endothelial expression of gp130 increased, and expression of gp130 in the lung was downregulated in the MWCNT-exposed group. We propose that pulmonary exposure to MWCNT can manifest as a reduced epithelial barrier and activator of vascular gp130-associated transsignaling that may promote susceptibility to cardiovascular derangements.

Mast cells represent a crucial cell type in host defense; however, maladaptive responses are contributing factors in the pathogenesis of allergic diseases. Previous work in our laboratory has shown that exposure to silver nanoparticles (AgNPs) results in mast cell degranulation via a non-immunoglobulin E (IgE) mechanism. In this study, we utilized a systems biology approach to identify novel genetic factors playing a role in AgNP-induced mast cell degranulation compared to the classical activation by antigen-mediated FceRI crosslinking. Mast cell degranulation was assessed in bone marrow-derived mast cells isolated from 23 strains of mice following exposure to AgNPs or FceRI crosslinking with dinitrophenyl (DNP). Utilizing strain-dependent mast cell degranulation, an association mapping study identified 3 chromosomal regions that were significantly associated with mast cell degranulation by AgNP and one non-overlapping region associated with DNP-mediated degranulation. Two of the AgNP-associated regions correspond to genes previously reported to be associated with allergic disorders (Trac2 on chromosome 1 and Traf6 on chromosome 2) and an uncharacterized gene identified on chromosome 1 (Fam126b). In conjunction, RNA-sequencing performed on mast cells from the high and low responder strains revealed 3754 and 34 differentially expressed genes that were unique to DNP and AgNP exposures, respectively. Select candidate genes include Ptger4, a gene encoding a G-protein coupled receptor in addition to a multifunctional adaptor protein, Txnip, that may be driving mast cell degranulation by AgNP. Taken together, we identified novel genes that have not been previously shown to play a role in nanoparticle-mediated mast cell activation. With further functional evaluation in the future, these genes may be potential therapeutic targets in the treatment of non-IgE mediated mast cell-linked disorders.