Investigation of pulmonary inflammatory responses following intratracheal instillation of and inhalation exposure to polypropylene microplastics.

BACKGROUND: Microplastics have been detected in the atmosphere as well as in the ocean, and there is concern about their biological effects in the lungs. We conducted a short-term inhalation exposure and intratracheal instillation using rats to evaluate lung disorders related to microplastics. We conducted an inhalation exposure of polypropylene fine powder at a low concentration of 2 mg/m3 and a high concentration of 10 mg/m3 on 8-week-old male Fischer 344 rats for 6 h a day, 5 days a week for 4 weeks. We also conducted an intratracheal instillation of polypropylene at a low dose of 0.2 mg/rat and a high dose of 1.0 mg/rat on 12-week-old male Fischer 344 rats. Rats were dissected from 3 days to 6 months after both exposures, and bronchoalveolar lavage fluid (BALF) and lung tissue were collected to analyze lung inflammation and lung injury. RESULTS: Both exposures to polypropylene induced a persistent influx of inflammatory cells and expression of CINC-1, CINC-2, and MPO in BALF from 1 month after exposure. Genetic analysis showed a significant increase in inflammation-related factors for up to 6 months. The low concentration in the inhalation exposure of polypropylene also induced mild lung inflammation. CONCLUSION: These findings suggest that inhaled polypropylene, which is a microplastic, induces persistent lung inflammation and has the potential for lung disorder. Exposure to 2 mg/m3 induced inflammatory changes and was thought to be the Lowest Observed Adverse Effect Level (LOAEL) for acute effects of polypropylene. However, considering the concentration of microplastics in a real general environment, the risk of environmental hazards to humans may be low.

Inflammogenic effect of polyacrylic acid in rat lung following intratracheal instillation.

BACKGROUND: Some organic chemicals are known to cause allergic disorders such as bronchial asthma and hypersensitivity pneumonitis, and it has been considered that they do not cause irreversible pulmonary fibrosis. It has recently been reported, however, that cross-linked acrylic acid-based polymer, an organic chemical, might cause serious interstitial lung diseases, including pulmonary fibrosis. We investigated whether or not intratracheal instillation exposure to cross-linked polyacrylic acid (CL-PAA) can cause lung disorder in rats. METHODS: Male F344 rats were intratracheally instilled with dispersed CL-PAA at low (0.2 mg/rat) and high (1.0 mg/rat) doses, and were sacrificed at 3 days, 1 week, 1 month, 3 months and 6 months after exposure to examine inflammatory and fibrotic responses and related gene expressions in the lungs. Rat lungs exposed to crystalline silica, asbestos (chrysotile), and NiO and CeO2 nanoparticles were used as comparators. RESULTS: Persistent increases in total cell count, neutrophil count and neutrophil percentage, and in the concentration of the cytokine-induced neutrophil chemoattractant (CINC)-1, CINC-2 and C-X-C motif chemokine 5 (CXCL5), which correlated with lung tissue gene expression, were observed in bronchoalveolar lavage fluid (BALF) from 3 days until at least 1 month following CL-PAA intratracheal instillation. Persistent increases in heme oxygenase-1 (HO-1) in the lung tissue were also observed from 3 days to 6 months after exposure. Histopathological findings of the lungs demonstrated that extensive inflammation at 3 days was greater than that in exposure to silica, NiO nanoparticles and CeO2 nanoparticles, and equal to or greater than that in asbestos (chrysotile) exposure, and the inflammation continued until 1 month. Fibrotic changes also progressed after 1 month postexposure. CONCLUSION: Our results suggested that CL-PAA potentially causes strong neutrophil inflammation in the rat and human lung.

Effect of Different Molecular Weights of Polyacrylic Acid on Rat Lung Following Intratracheal Instillation.

BACKGROUND: We conducted intratracheal instillations of different molecular weights of polyacrylic acid (PAA) into rats in order to examine what kinds of physicochemical characteristics of acrylic acid-based polymer affect responses in the lung. METHODS: F344 rats were intratracheally exposed to a high molecular weight (HMW) of 598 thousand g/mol or a low molecular weight (LMW) of 30.9 thousand g/mol PAA at low and high doses. Rats were sacrificed at 3 days, 1 week, 1 month, 3 months and 6 months post exposure. RESULTS: HMW PAA caused persistent increases in neutrophil influx, cytokine-induced neutrophil chemoattractants (CINC) in the bronchoalveolar lavage fluid (BALF), and heme oxygenase-1 (HO-1) in the lung tissue from 3 days to 3 months and 6 months following instillation. On the other hand, LMW PAA caused only transient increases in neutrophil influx, CINC in BALF, and HO-1 in the lung tissue from 3 days to up to 1 week or 1 month following instillation. Histopathological findings of the lungs demonstrated that the extensive inflammation and fibrotic changes caused by the HMW PAA was greater than that in exposure to the LMW PAA during the observation period. CONCLUSION: HMW PAA induced persistence of lung disorder, suggesting that molecular weight is a physicochemical characteristic of PAA-induced lung disorder.

Crosslinked Structure of Polyacrylic Acid Affects Pulmonary Fibrogenicity in Rats.

We conducted intratracheal instillations of polyacrylic acid (PAA) with crosslinking and non-crosslinking into rats in order to examine what kinds of physicochemical characteristics of acrylic-acid-based polymers affect responses in the lung. F344 rats were intratracheally exposed to similar molecular weights of crosslinked PAA (CL-PAA) (degree of crosslinking: ~0.1%) and non-crosslinked PAA (Non-CL-PAA) at low and high doses. Rats were sacrificed at 3 days, 1 week, 1 month, 3 months, and 6 months post-exposure. Both PAAs caused increases in neutrophil influx, cytokine-induced neutrophil chemoattractants (CINC) in the bronchoalveolar lavage fluid (BALF), and heme oxygenase-1 (HO-1) in the lung tissue from 3 days to 6 months following instillation. The release of lactate dehydrogenase (LDH) activity in the BALF was higher in the CL-PAA-exposed groups. Histopathological findings of the lungs demonstrated that the extensive fibrotic changes caused by CL-PAA were also greater than those in exposure to the Non-CL- PAA during the observation period. CL-PAA has more fibrogenicity of the lung, suggesting that crosslinking may be one of the physicochemical characteristic factors of PAA-induced lung disorder.

The charge reduction rate for multiply charged polymer ions via ion-ion recombination at atmospheric pressure.

The charge reduction of multiply charged macromolecular ions via recombination with small ions in the gas phase is commonly employed to modulate the charge on macromolecules prior to mass spectrometric and mobility analyses. We employ a recently developed continuum-Molecular Dynamics (MD) calculation approach to determine the recombination rate coefficient of multiply charged (1 to 7 excess positive charged) polyethylene glycol ions (mass of 4600 Da) with smaller singly charged anions, modeled as NO2- ions. The continuum-MD approach accounts explicitly for the influence of the background gas on the recombination process, accounts explicitly for ion translational, vibrational, and rotational motion, and enables recombination rate coefficient calculation in nitrogen near atmospheric pressure, wherein neither low pressure nor high pressure recombination theories are strictly applicable. Continuum-MD simulations yield recombination rate coefficients near 3.9 × 10-14 m3 s-1 for singly charged ions, increasing to 3.0 × 10-11 m3 s-1 for the +7 ion. Pre-existing collision rate coefficient expressions for rigid ions are found to be within a factor of 2-5 of calculations for all charge states, but their use requires knowledge of an appropriate collision distance, which is not well-defined for flexible polymer ions. Continuum-MD-inferred rate coefficients are incorporated into a model of charge reduction, and the charge state distribution versus anion concentration determined with it is compared to charge reduction measurements made via atmospheric pressure differential mobility analysis. Good agreement is observed between simulations and experiments; although results are extremely sensitive to the recombination rate coefficients, experimental results are bound by models utilizing rates within a factor of 2 (0.5-2.0×) of the continuum-MD rates.

Calculation of the ion-ion recombination rate coefficient via a hybrid continuum-molecular dynamics approach.

Accurate calculation of the ion-ion recombination rate coefficient has been of long-standing interest as it controls the ion concentration in gas phase systems and in aerosols. We describe the development of a hybrid continuum-molecular dynamics (MD) approach to determine the ion-ion recombination rate coefficient. This approach is based on the limiting sphere method classically used for transition regime collision phenomena in aerosols. When ions are sufficiently far from one another, the ion-ion relative motion is described by diffusion equations, while within a critical distance, MD simulations are used to model ion-ion motion. MD simulations are parameterized using the Assisted Model Building with Energy Refinement force-field as well as by considering partial charges on atoms. Ion-neutral gas collisions are modeled in two mutually exclusive cubic domains composed of 103 gas atoms each, which remain centered on the recombining ions throughout calculations. Example calculations are reported for NH4 + recombination with NO2 - in He, across a pressure range from 10 kPa to 10 000 kPa. Excellent agreement is found in comparison with calculations to literature values for the 100 kPa recombination rate coefficient (1.0 × 10-12 m3 s-1) in He. We also recover the experimentally observed increase in the recombination rate coefficient with pressure at sub-atmospheric pressures, and the observed decrease in the recombination rate coefficient in the high pressure continuum limit. We additionally find that non-dimensionalized forms of rate coefficients are consistent with recently developed equations for the dimensionless charged particle-ion collision rate coefficient based on Langevin dynamics simulations.

Simultaneous ion and neutral evaporation in aqueous nanodrops: experiment, theory, and molecular dynamics simulations.

We use a combination of tandem ion mobility spectrometry (IMS-IMS, with differential mobility analyzers), molecular dynamics (MD) simulations, and analytical models to examine both neutral solvent (H2O) and ion (solvated Na(+)) evaporation from aqueous sodium chloride nanodrops. For experiments, nanodrops were produced via electrospray ionization (ESI) of an aqueous sodium chloride solution. Two nanodrops were examined in MD simulations: a 2500 water molecule nanodrop with 68 Na(+) and 60 Cl(-) ions (an initial net charge of z = +8), and (2) a 1000 water molecule nanodrop with 65 Na(+) and 60 Cl(-) ions (an initial net charge of z = +5). Specifically, we used MD simulations to examine the validity of a model for the neutral evaporation rate incorporating both the Kelvin (surface curvature) and Thomson (electrostatic) influences, while both MD simulations and experimental measurements were compared to predictions of the ion evaporation rate equation of Labowsky et al. [Anal. Chim. Acta, 2000, 406, 105-118]. Within a single fit parameter, we find excellent agreement between simulated and modeled neutral evaporation rates for nanodrops with solute volume fractions below 0.30. Similarly, MD simulation inferred ion evaporation rates are in excellent agreement with predictions based on the Labowsky et al. equation. Measurements of the sizes and charge states of ESI generated NaCl clusters suggest that the charge states of these clusters are governed by ion evaporation, however, ion evaporation appears to have occurred with lower activation energies in experiments than was anticipated based on analytical calculations as well as MD simulations. Several possible reasons for this discrepancy are discussed.

Intratracheal instillation of polyacrylic acid induced pulmonary fibrosis with elevated transforming growth factor-ß1 and connective tissue growth factor.

We investigated the intratracheal instillation of Polyacrylic acid (PAA) in rats to determine if it would cause pulmonary disorders, and to see what factors would be associated with the pathological changes. Male F344 rats were intratracheally instilled with low (0.2 mg/rat) and high (1.0 mg/rat) doses of PAA. They were sacrificed at 3 days, 1 week, 1 month, 3 months, and 6 months after PAA exposure to examine inflammatory and fibrotic changes in the lungs. There was a persistent increase in the neutrophil count, lactate dehydrogenase (LDH) levels, cytokine-induced neutrophil chemoattractant (CINC) values in bronchoalveolar lavage fluid (BALF), and heme oxygenase-1 (HO-1) in lung tissue. Transforming growth factor-beta 1 (TGF-ß1), a fibrotic factor, showed a sustained increase in the BALF until 6 months after intratracheal instillation, and connective tissue growth factor (CTGF) in lung tissue was elevated at 3 days after exposure. Histopathological findings in the lung tissue showed persistent (more than one month) inflammation, fibrotic changes, and epithelial-mesenchymal transition (EMT) changes. There was also a strong correlation between TGF-ß1 in the BALF and, especially, in the fibrosis score of histopathological specimens. Intratracheal instillation of PAA induced persistent neutrophilic inflammation, fibrosis, and EMT in the rats' lungs, and TGF-ß1 and CTGF appeared to be associated with the persistent fibrosis.

Impact of increased reimbursement for ambulance transportation on hospital acceptance in Japan: a difference-in-difference study.

OBJECTIVE: Emergency medical services (EMS) often face difficulties in finding accepting hospitals in Japan. The universal medical insurance system in Japan increased the reimbursement for ambulance transportation acceptance at night, and on Sundays and holidays from 1 April 2016. This study investigated the effect of the reimbursement increase on the number of EMS calls, and transportation time from arrival at the scene to arrival at the hospital. DESIGN: A difference-in-difference study. The treatment group consisted of people who called an ambulance at night while the control group consisted of people who called an ambulance during the daytime. SETTING: The national ambulance records of the Fire and Disaster Management Agency in Japan from 1 April 2015 to 31 December 2016. PARTICIPANTS: 7 625 463 ambulance dispatches were eligible for inclusion. PRIMARY AND SECONDARY OUTCOME MEASURES: The changes in EMS calls, transportation time and the number of ambulance transports per 1000 population in one month in a comparison of daytime and night-time transport. RESULTS: The treatment effect (night-time vs daytime) on the number of EMS calls was -0.013 (95% CI, -0.023 to -0.004), which was significant. The transportation time decreased slightly by 0.080 min (95% CI, -0.157 to -0.004). No impact was observed on the number of ambulance transports per 1000 population per month (0.00; 95% CI, -0.008 to 0.002). CONCLUSION: An increase in reimbursement for ambulance transportation acceptance was associated with a decrease in the number of EMS calls. Further strategies for decreasing the number of EMS calls are needed to avoid delays in the treatment of emergency patients with critical illness.

Measuring the effects of respiratory protective equipment and other protectors in preventing the scattering of vocalization droplets.

This study was conducted to quantitatively examine the effects of respiratory protective equipment (respirators) and various other types of protectors in preventing the scattering of vocalization droplets. Each of 12 adult male volunteers was asked to vocalize intermittently for 1 min at a target intensity of approximately 100 dBA in an experimental room adjusted to a humidity of approximately 60-70%. The subjects vocalized while wearing respirators, other types of protectors, or no protectors at all. The droplet concentration in a particle size range of 0.3 to 10 µm was measured under each experimental condition, and the transmitted particle concentration and penetration were calculated. The concentration and penetration of particles transmitted from the respirators were lower than those transmitted from the other protectors examined. The probability of infection reduction through the use of the protectors was estimated from the data obtained on the effectiveness of the protectors in preventing the scattering of droplets. We concluded that there is no need for additional droplet scattering prevention in various work settings when appropriate respirators are used under optimal conditions.

Prevertebral abscess associated with meningitis: Double cause of neck stiffness.

While neck stiffness belongs to the classic triad of meningitis symptoms together with fever and altered mental status, it can also be attributed to inflammation from prevertebral space infection. We describe a difficult-to-diagnose case of prevertebral abscess associated with meningitis. Prevertebral space infection, vertebral osteomyelitis, and meningitis are reported to be associated with each other. When a patient presents with an altered mental status due to meningitis, signs and symptoms may be obscured and physicians may be unable to conduct detailed physical examinations or identify symptoms beyond neck stiffness. The threshold for imaging evaluation may need to be lowered for patients at high risk for prevertebral abscess or vertebral osteomyelitis. Physicians need to recognize this clinical entity, as prompt referral to specialists in head and neck surgery is essential for timely drainage.

Pulmonary toxicity of tungsten trioxide nanoparticles in an inhalation study and an intratracheal instillation study.

OBJECTIVES: We conducted inhalation and intratracheal instillation studies in order to examine the effects of tungsten trioxide (WO3 ) nanoparticles on the lung, and evaluated whether or not the nanoparticles would cause persistent lung inflammation. METHODS: In the inhalation study, male 10-week-old Fischer 334 rats were classified into 3 groups. The control, low-dose, and high-dose groups inhaled clean air, 2, and 10 mg/m3 WO3 nanoparticles, respectively, for 6 h each day for 4 weeks. The rats were dissected at 3 days, 1 month, and 3 months after the inhalation, and the bronchoalveolar lavage fluid (BALF) and lung tissue were examined. In the intratracheal instillation study, male 12-week-old Fischer 334 rats were divided into 3 subgroups. The control, low-dose, and high-dose groups were intratracheally instilled 0.4 ml distilled water, 0.2, and 1.0 mg WO3 nanoparticles, respectively, dissolved in 0.4 ml distilled water. The rats were sacrificed at 3 days, 1 week, and 1 month after the intratracheal instillation, and the BALF and lung tissue were analyzed as in the inhalation study. RESULTS: The inhalation and instillation of WO3 nanoparticles caused transient increases in the number and rate of neutrophils, cytokine-induced neutrophil chemoattractant (CINC)-1, and CINC-2 in BALF, but no histopathological changes or upregulation of heme oxygenase (HO)-1 in the lung tissue. CONCLUSION: Our results suggest that WO3 nanoparticles have low toxicity to the lung. According to the results of the inhalation study, we also propose that the no observed adverse effect level (NOAEL) of WO3 nanoparticles is 2 mg/m3 .

Pulmonary disorder induced by cross-linked polyacrylic acid.

OBJECTIVES: Organic polymers are materials widely used in our daily lives, such as daily necessities, foods, and medicines. There have been reports recently that cross-linked polyacrylic acid (CL-PAA) can possibly cause serious lung disease. We investigated whether intratracheal instillation of CL-PAA causes pulmonary disorder in rats. METHODS: Male F344 rats were administered low (0.2 mg/rat) and high (1.0 mg/rat) doses of CL-PAA intratracheally and were dissected 3 days, 1 week, 1 month, 3 months, and 6 months after exposure to examine inflammatory and fibrotic responses in the lungs. Only the high-dose specimens were subjected to ultrasonic dispersion treatment of the administered material. RESULTS: There was a dose-dependent increase in the total cell count, neutrophil count, neutrophil percentage, lactate dehydrogenase (LDH), surfactant protein D (SP-D), cytokine-induced neutrophil chemoattractant (CINC)-1 and CINC-2 values in bronchoalveolar lavage fluid (BALF) from 3 days to at least 3 months after intratracheal administration of CL-PAA. Heme oxygenase-1 (HO-1) in lung tissue was also persistently elevated from 3 days to 6 months after exposure. Alkaline phosphatase (ALP) in BALF was elevated at 3 days and 1 month after exposure only in the high-dose group. Histopathological findings in lung tissue showed inflammatory and fibrotic changes from 3 days after administration, and we observed obvious inflammatory changes for up to 3 months and fibrotic changes for up to 6 months. CONCLUSION: Intratracheal administration of CL-PAA induced persistent neutrophilic inflammation and fibrosis in the rats' lungs, suggesting that CL-PAA may have inflammogenic and fibrogenic effects.

Factors associated with difficulty of hospital acceptance of patients suspected to have cerebrovascular diseases: A nationwide population-based observational study.

Although it is essential to shorten the interval to initial treatment in the care of acute ischemic stroke, some hospitals in Japan reject requests for hospital acceptance from on-scene emergency medical service personnel because of limited resources, which can cause delays in care. We aimed to assess the risk factors for difficulty of hospital acceptance of patients suspected to have cerebrovascular diseases. We conducted a retrospective analysis of the national ambulance records of the Fire and Disaster Management Agency in Japan in 2016. Multivariable logistic regression analysis was used to assess the association between difficulty of hospital acceptance of patients suspected to have cerebrovascular diseases and prehospital factors. During the study period, a total of 222,926 patients were included, and 5283 patients (2.4%) experienced difficulties in hospital acceptance. In multivariable analysis, nights (adjusted odds ratio [AOR] 1.54, 95% confidence interval [CI] 1.45-1.64), weekends (AOR 1.32, 95% CI 1.24-1.40), <25 percentile ratio of emergency physicians and neurosurgeons to all physicians (AOR 1.13, 95% CI 1.03-1.23) (AOR 1.36, 95% CI 1.25-1.48), and mean age of physicians (AOR 1.06, 95% CI 1.05-1.07) were significantly associated with difficulties of hospital acceptance of patients suspected to have cerebrovascular disease. There was a marked regional variation in the difficulties of hospital acceptance. Among the national ambulance records of patients suspected to have cerebrovascular diseases, certain prehospital factors such as weekends were positively associated with difficulty of hospital acceptance. A comprehensive strategy for hospital acceptance of patients with cerebrovascular diseases considering regional variation is required.

The Dedicated Emergency Physician Model of emergency care is associated with reduced pre-hospital transportation time: A retrospective study with a nationwide database in Japan.

In Japan, the increasing number of patients needing emergency medical care due to population aging is a major public health problem. Recently, emergency medicine in Japan has seen a growth in the number of Dedicated Emergency Physician Model style departments. We aimed to determine whether there is an association between Dedicated Emergency Physician Model emergency care and pre-hospital transportation time. We conducted a secondary analysis of a Japanese national pre-hospital database from 2010 to 2014. Three regions (group 1: Urayasu city and Ichikawa city in Chiba prefecture, group 2: Kamakura city, Chigasaki city, Fujisawa city and Zushi city in Kanagawa prefecture, and group 3: Fukui city in Fukui prefecture) were evaluated as Dedicated Emergency Physician Model emergency medicine areas. We compared transportation times in these areas with all municipalities in the same prefectures, and with a nearby area using multivariate linear regression with cluster adjustment. The variables used for adjustment are the time from Emergency Medical Services activation to the scene, month, day of the month, day of the week, time of day, age, gender, type of injury, severity, and location of call. Compared with all municipalities in each prefecture there were significant reductions in pre-hospital transportation time: 4.2 minutes (95% confidence interval, 0.9 to 7.5, p<0.05) in Group 1, 6.2 minutes (95%CI, 2.9 to 9.6, p<0.01) fin Group 2 and 7.5 minutes (95%CI, 6.0 to 9.0, p<0.01) in Group 3. Compared with nearby areas, there were statistically significant reductions in transportation time in Group 1, 6.8 minutes (95%CI, 0.7 to 12.8, p<0.05) and in Group 2, 6.8 minutes (95%CI, 3.7 to 9.9, p<0.05). There was a trend for reduced transportation time in Group 3, 2.3 minutes, (5.3 to -0.6, p<0.1). Areas with a Dedicated Emergency Physician Model are associated with reduced pre-hospital transportation time.