Methacholine induced airway contraction in porcine precision cut lung slices from indoor and outdoor reared pigs.

Repetitive exposure to bioaerosols in swine production facilities (SPF) promotes respiratory dysfunction in workers and animals. An adequate understanding of the impact of the SPF environment on pulmonary physiology is needed. However, there is currently no sufficient ex vivo model to investigate the cause for agriculture-related lung disease. The precision cut lung slices (PCLS) technique represents a practical and useful procedure for ex vivo studies. Our goal was to use the PCLS technique to develop a model of agriculture-related lung diseases using a physiologically relevant animal model, the domesticated pig. Freshly prepared pig lung tissue cores were sectioned into 300 µm slices and viability was measured by lactate dehydrogenase activity and live/dead staining. Airway contractility in response to a methacholine (MCh) dose gradient (10-7-10-4 M) was measured. After the last MCh dose, PCLS were incubated with 1 mM chloroquine to allow airways to relax. Time-lapse images were taken every minute for 35 minutes and used to determine airway lumen area changes. Porcine PCLS remained viable and demonstrated metabolic activity for three days. PCLS from indoor and outdoor pigs contracted in response to MCh exposure and relaxed when incubated with chloroquine. Notably, PCLS of indoor pigs showed greater airway constriction in response to 10-5 M MCh exposure compared to outdoor pig PCLS (P<0.05). These data suggest that exposure to the indoor pig production environment may be associated with hyperresponsiveness in swine airways, and support future studies to investigate lung response to inflammatory substances using the porcine PCLS model.

Profiling of cell stress proteins reveals decreased expression of enzymatic antioxidants in tracheal epithelial tissue of pigs raised indoors.

Exposure to indoor swine production facilities (SPF) environments causes airway inflammation and diseases including asthma, chronic bronchitis and chronic obstructive pulmonary disease (COPD) in facility workers. However, less is known about the impact of SPF exposures on the respiratory health of pigs. Respiratory symptoms are associated with repeat exposure to SPF, specifically inhalation of organic dust and other air pollutants therein. A thorough understanding of the molecular pathways regulated by SPF exposure is needed to understand airway inflammation and chronic inflammatory lung disease. The present study measured the expression of proteins associated with oxidative stress and antioxidant defenses in the tracheal epithelial tissues of pigs reared in SPF or on pasture. Proteome profiler cell stress arrays, western blotting and enzyme activity assays were utilized to measure protein expression and activity levels in tracheal epithelial tissue extracts of pigs. It was determined that pigs raised in SPF express significantly less enzymatic antioxidants, including superoxide dismutase (SOD), within their tracheal epithelial tissues compared to pasture raised pigs. Concomitantly, tracheal epithelial tissues of SPF raised pigs had lower SOD and catalase antioxidant activity levels compared to pasture raised pigs. The observations summarized herein provide evidence that exposure to swine production environments influence endogenous enzymatic antioxidant defenses within the tracheal epithelial tissues of pigs. This study offers insight for understanding the effect of continuous exposure to SPF pollutants on endogenous antioxidant defenses in the airway epithelial and may be helpful in understanding human airway responses to swine barn exposures.

Sorrel Extract Reduces Oxidant Production in Airway Epithelial Cells Exposed to Swine Barn Dust Extract In Vitro.

Exposure to hog barn organic dust contributes to occupational lung diseases, which are mediated by inflammatory and oxidative stress pathways. Isoprostanes-a family of eicosanoids produced by oxidation of phospholipids by oxygen radicals-are biomarkers of pulmonary oxidative stress. Importantly, 8-isoprostane has been implicated as a key biomarker and mediator of oxidative stress because it is a potent pulmonary vasoconstrictor. Antioxidants found in fruits and vegetables hold promise for preventing or reducing effects of oxidative stress-related diseases including chronic bronchitis and chronic obstructive pulmonary disease (COPD). Here, we investigated 8-isoP and oxidant production by organic dust-exposed airway epithelial cells and the inhibitory effects of an extract from calyces of the sorrel plant, Hibiscus sabdariffa, on oxidant-producing pathways. Confluent cultures of normal human tracheobronchial epithelial cells were pretreated or not with 1% sorrel extract prior to 5% dust extract (DE) exposure. Following DE treatments, live cells, cell-free supernatants, or cell extracts were evaluated for the presence of 8-isoprostane, superoxide, hydrogen peroxide, nitric oxide, hydroxyl radical, peroxynitrite, and catalase activity to evaluate sorrel's inhibitory effect on oxidative stress. The well-known radical scavenging antioxidant, N-acetyl cysteine (NAC), was used for comparisons with sorrel. DE exposure augmented the production of all radicals measured including 8-isoprostane (p value < 0.001), which could be inhibited by NAC or sorrel. Among reactive oxygen and nitrogen species generated in response to DE exposure, sorrel had no effect on H2O2 production and NAC had no significant effect on NO· production. The observations reported here suggest a possible role for sorrel in preventing 8-isoprostane and oxidant-mediated stress responses in bronchial epithelial cells exposed to hog barn dust. These findings suggest a potential role for oxidative stress pathways in mediating occupational lung diseases and antioxidants within sorrel and NAC in reducing dust-mediated oxidative stress within the airways of exposed workers.

Farm animal models of organic dust exposure and toxicity: insights and implications for respiratory health.

PURPOSE OF REVIEW: Modern food animal production is a major contributor to the global economy, owing to advanced intensive indoor production facilities aimed at increasing market readiness and profit. Consequences of these advances are accumulation of dusts, gases, and microbial products that diminish air quality within production facilities. Chronic inhalation exposure contributes to onset and exacerbation of respiratory symptoms and diseases in animals and workers. This article reviews literature regarding constituents of farm animal production facility dusts, animal responses to production building and organic dust exposure, and the effect of chronic inhalation exposure on pulmonary oxidative stress and inflammation. RECENT FINDINGS: Porcine models of production facility and organic dust exposures reveal striking similarities to observations of human cells, tissues, and clinical data. Oxidative stress plays a key role in mediating respiratory diseases in animals and humans, and enhancement of antioxidant levels through nutritional supplements can improve respiratory health. SUMMARY: Pigs are well adapted to the exposures common to swine production buildings and thus serve as excellent models for facility workers. Insight for understanding mechanisms governing organic dust associated respiratory diseases may come from parallel comparisons between farmers and the animals they raise.

Effect of Mucin and Bicarbonate Ion on Corrosion Behavior of AZ31 Magnesium Alloy for Airway Stents.

The biodegradable ability of magnesium alloys is an attractive feature for tracheal stents since they can be absorbed by the body through gradual degradation after healing of the airway structure, which can reduce the risk of inflammation caused by long-term implantation and prevent the repetitive surgery for removal of existing stent. In this study, the effects of bicarbonate ion (HCO3-) and mucin in Gamble's solution on the corrosion behavior of AZ31 magnesium alloy were investigated, using immersion and electrochemical tests to systematically identify the biodegradation kinetics of magnesium alloy under in vitro environment, mimicking the epithelial mucus surfaces in a trachea for development of biodegradable airway stents. Analysis of corrosion products after immersion test was performed using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX) and X-ray diffraction (XRD). Electrochemical impedance spectroscopy (EIS) was used to identify the effects of bicarbonate ions and mucin on the corrosion behavior of AZ31 magnesium alloys with the temporal change of corrosion resistance. The results show that the increase of the bicarbonate ions in Gamble's solution accelerates the dissolution of AZ31 magnesium alloy, while the addition of mucin retards the corrosion. The experimental data in this work is intended to be used as foundational knowledge to predict the corrosion behavior of AZ31 magnesium alloy in the airway environment while providing degradation information for future in vivo studies.

Synthesis, structure, and biocompatibility of pulsed laser-deposited TiN nanowires for implant applications.

A bottom-up based pulsed laser deposition technique has been used to grow titanium nitride (TiN) nanowires on single crystalline substrates. The first step of this method is the dissolution of laser ablated gaseous TiN material in the nanodimensional catalytic gold (Au) liquid islands located on the substrate surfaces. The continuous dissolution of TiN results in the supersaturation of liquid Au with TiN followed by extrusion of solid TiN material in the nanowire form at the liquid/solid interface. The growth of TiN nanowires continues as long as its dissolution rate into the catalyst Au matches the extrusion rate of solid TiN. This bottom-up approach gives rise to a one-dimensional TiN nanowire structures (length: 200-300 nm and diameter: 20-30 nm) capped with Au. The ascent of Au nanodots to the top of TiN nanowires can be explained based on breaking of weaker bonds and building of stronger bonds. The TiN nanowires are provided vertical alignment by selecting a plane of the substrate that provides the least lattice mismatching to the (111) plane of TiN which has lower surface energy than its other planes: (100) or (110). After the successful formation and structural characterization, a lactate dehydrogenase (LDH) release assay has been used to confirm the biocompatibility and cytotoxicity of these nanowires.

A Method for Isolation of Pasteuria penetrans Endospores for Bioassay and Genomic Studies.

A rapid method for collection of Pasteuria penetrans endospores was developed. Roots containing P. penetrans-infected root-knot nematode females were softened by pectinase digestion, mechanically processed, and filtered to collect large numbers of viable endospores. This method obviates laborious handpicking of Pasteuria-infected females and yields endospores competent to attach to and infect nematodes. Endospores are suitable for morphology studies and DNA preparations.