Autopsy is recognized as the gold standard for examining the body of the deceased. However postmortem inspection, which is a method for examining the body through non-invasive modalities, is very limited. Therefore, several methods have been studied to complement postmortem examination, and endoscopy emerged as a possible method. The authors of the present study performed endoscopic examination before autopsy, and the endoscopic findings were confirmed by subsequent autopsy. The endoscope was inserted mainly through the nostril due to the rigor mortis of the jaw joint. The pharynx, larynx, trachea, and esophagus were examined. Endoscopic examination was performed on a total of 35 cases. Endoscopy revealed froth in the airway (drowning cases), as well as the presence of thermal denaturation of structures in the airway and soot attached to the froth in the airway (fire death cases). However, relevant findings were not noted during external examination. In addition, agrochemical substances were detected in the airway (agrochemical poisoning death cases) during endoscopic examination. The study found that useful information can be obtained for investigating the death and estimating postmortem interval through endoscopic examination. It is thought that minimally invasive autopsy procedures, including endoscopy, cannot replace conventional autopsies, but can be used as adjuncts instead. In particular, minimally invasive autopsy procedures are thought to be useful for postmortem inspection. To this end, continued studies, as well as the development of equipment suited for postmortem examination, are needed.
Autopsy* ; Complement System Proteins ; Endoscopes ; Endoscopy ; Esophagus ; Jaw ; Joints ; Larynx ; Methods ; Minimally Invasive Surgical Procedures ; Pharynx ; Poisoning ; Rigor Mortis ; Soot ; Trachea

OBJECTIVETo explore the carbon black induced effects of lung morphology and pro-inflammation in mice, based on the carbon black aerosol dynamic inhalation exposure model.

METHODSThe carbon black aerosol generated by dynamic inhalation device was imported exposure chamber to mice. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the characters of carbon black. Sixty 9-week-old male BALB/c mice were randomly divided into two control groups, 7 d exposure group and 14 d exposure group. The numbers of four groups of animals were 15, respectively. Mice were exposed to carbon black in the inhalation chamber at (29.33 ± 9.10) mg/m(3) for 6 h/d for continuous exposure 7 d and 14 d, respectively. After 7 d and 14 d exposure, the mice were sacrificed after the last exposure for 24 h. Control mice were killed at 7 d and 14 d. The trachea, lungs, liver, kidneys, and spleen tissues were separated and weighted. Hematoxylin and eosin (HE) staining was used to observe pathological changes of lung by light microscopy. Pulmonary interleukin-8 (IL-8) expression was analyzed by immunohistochemistry. Transmission electron microscopy was used to observe the ultra structure of lung tissue.

RESULTSAfter 14 d exposure carbon black, the lung coefficient was increased in exposure group compared with control (0.61 ± 0.03 vs 0.79 ± 0.06, t = 6.26, P < 0.01). The spleen coefficient were higher than control(0.39 ± 0.04 vs 0.51 ± 0.06, t = 4.23, P < 0.01) . Other organ coefficients were no significant difference between CB group and control group.Histopathology displayed carbon black particles were deposited in the alveoli and lung bronchial wall in 7 d and 14 d groups. The black carbon particles were deposited within the lung tissue of mice in 14 d group. There were cilia damage, serious damage to the alveolar wall, inflammatory cell infiltration and more hyperemia in 14 d group. Immunohistochemistry showed the level of IL-8 in 7 d (0.272 ± 0.011) and 14 d (0.422 ± 0.065) exposure group were higher than control group in 14 d (0.188 ± 0.041) , F = 31.89, P < 0.01. TEM showed that the lung tissue vision was clear and organelle integrity in the control group. The particles appeared in lung tissue macrophage lysosomes in exposure group, the electron density was consistent with the carbon black particles.

CONCLUSIONThe dynamic carbon black particles exposure can affect the lung and spleen coefficient, damage integrity of lung morphology and induce inflammation in mice.

Aerosols ; Animals ; Cilia ; Inflammation ; Inhalation Exposure ; Interleukin-8 ; Lung ; Macrophages, Alveolar ; Male ; Mice ; Mice, Inbred BALB C ; Pulmonary Alveoli ; Soot ; Spleen

The contribution of particles to cardiovascular mortality and morbidity has been enlightened by epidemiologic and experimental studies. However, adverse biological effects of the particles with different sizes on cardiovascular cells have not been well recognized. In this study, sub-cultured human umbilical vein endothelial cells (HUVECs) were exposed to increasing concentrations of pure quartz particles (DQ) of three sizes (DQPM1, <1 μm; DQPM3-5, 3-5 μm; DQPM5, 5 μm) and carbon black particles of two sizes (CB0.1, <0.1 μm; CB1, <1 μm) for 24 h. Cytotoxicity was estimated by measuring the activity of lactate dehydrogenase (LDH) and cell viability. Nitric oxide (NO) generation and cytokines (TNF-α and IL-1β) releases were analyzed by using NO assay and enzyme-linked immunoabsorbent assay (ELISA), respectively. It was found that both particles induced adverse biological effects on HUVECs in a dose-dependent manner. The size of particle directly influenced the biological activity. For quartz, the smaller particles induced stronger cytotoxicity and higher levels of cytokine responses than those particles of big size. For carbon black particles, CB0.1 was more capable of inducing adverse responses on HUVECs than CB1 only at lower particle concentrations, in contrast to those at higher concentrations. Meanwhile, our data also revealed that quartz particles performed stronger cell damage and produced higher levels of TNF-α than carbon black particles, even if particles size was similar. In conclusion, particle size as well as particle composition should be both considered in assessing vascular endothelial cells injury and inflammation responses induced by particles.
Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Interleukin-1beta ; secretion ; L-Lactate Dehydrogenase ; metabolism ; Nitric Oxide ; biosynthesis ; Particle Size ; Particulate Matter ; chemistry ; pharmacology ; Quartz ; chemistry ; Soot ; chemistry ; Time Factors ; Tumor Necrosis Factor-alpha ; secretion

Ambient air pollution, including particulate matter (PM) and gaseous pollutants, represents an important environmental factor that adversely affects human health. PM is a complex mixture of extremely small particles and liquid droplets. Particle pollution is made up of a number of components, including acids (such as nitrates and sulfates), organic chemicals, metals, and soil or dust particles. Subtypes of atmospheric PM include suspended particulate matter, respirable suspended particles (particles with a diameter of 10 microm or less), fine particles (diameter of 2.5 microm or less), ultrafine particles, and soot. Sources of particulate matter can be artificial or natural. The effects of inhaling PM that have been widely studied in humans and animals now include asthma, lung cancer, cardiovascular mortality, respiratory diseases, birth defects, and premature death. This review focus on the impact of PM on health outcomes such as respiratory disease, heart disease, and cancer.
Air Pollution ; Animals ; Asthma ; Congenital Abnormalities ; Dust ; Heart Diseases ; Humans ; Inhalation ; Lung Neoplasms ; Metals ; Mortality ; Mortality, Premature ; Nitrates ; Organic Chemicals ; Particulate Matter* ; Soil ; Soot

OBJECTIVES: This study was conducted to determine whether nano-sized carbon black exposure results in greater damage in high fat diet-induced overweight rats than normal weight ones and to identify the possible causes of any differences. METHODS: Two groups of Sprague-Dawley rats allocated by body weight (normal and overweight) were exposed to aerosolized nano-sized carbon black for 6 hours a day, 5 days per week over a 4-week period. Differential cell counts, lactate dehydrogenase (LDH) activities and albumin concentrations were measured in bronchoalveolar lavage (BAL) fluid, and histopathological findings in the lungs were evaluated. Tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-6 were measured in BAL fluid and supernatants of lipopolysaccharide(LPS)-stimulated lymphocyte culture. RESULTS: Rats exposed to high concentrations of nano-sized carbon black showed significantly increased (p<0.05) polymorphonuclear leukocyte number and LDH activity in the BAL fluid from both overweight and normal rats. Mild histopathological changes were observed in normal rats irrespective of carbon black concentrations. However, severe histological scores were found in overweight rats (1.75+/-0.46, 2.25+/-0.46, and 2.88+/-0.35 after low, medium, and high concentration exposures). Proinflammatory cytokine levels of TNF-alpha and IL-6 were significantly higher in the supernatant of LPS-stimulated lymphocytes of overweight rats, whereas there was no significant difference in the BAL fluid between normal and overweight rats. CONCLUSIONS: Inflammation and damage to lungs exposed to nano-sized carbon black was more severe in high fat diet-induced overweight rats compared to normal rats.
Animals ; Body Weight ; Bronchoalveolar Lavage ; Carbon* ; Cell Count ; Inflammation ; Inhalation ; Interleukin-6 ; Interleukins ; L-Lactate Dehydrogenase ; Lung* ; Lymphocytes ; Neutrophils ; Obesity ; Overweight* ; Rats* ; Rats, Sprague-Dawley ; Soot* ; Tumor Necrosis Factor-alpha

OBJECTIVETo investigate the protein expression of caveolin-1 in type II alveolar epithelial cells (A549) exposed to carbon black nanoparticles (CB NPs) and the role of caveolin in the endocytosis of CB NPs.

METHODSA549 cells were exposed to 0, 25, 50, 100, 200, and 400 µg/ml CB NPs for 24 h; then, trypan blue assay was applied to determine the cell viability. A549 cells were also exposed to 0, 25, 50, and 100 µg/ml CB NPs for 24 h, then, transmission electron microscopy (TEM) and flow cytometry were applied to observe the morphological change of cells and cellular side scatter (SSC), and Western blot was used to analyze the effect of CB NPs on the protein expression of caveolin-1. A549 cells were co-exposed to1 µg/ml filipin and 100 µg/ml CB NPs for 24 h, then, the cellular SSC was observed.

RESULTSCompared with controls, the A549 cells exposed to 200 and 400 µg/ml CB NPs had the cell viability decreased by 38.2% and 46.6%, respectively (P < 0.05), while those exposed to 25, 50, and 100 µg/ml CB NPs showed no significant decrease in cell vitality (P > 0.05). The protein expression of caveolin-1 was significantly higher in the cells exposed to 50 and 100 µg/ml CB NPs than in controls (P < 0.05). The TEM showed that plasmalemmal vesicles containing black particles were found in the cytoplasm of the cells exposed to 50 and 100 µg/ml CB NPs. The flow cytometry showed that the cellular SSC ratio increased from 1.007 to 1.331 as the dose of CB NPs rose within 0 ∼ 100 µg/ml and fell to 1.25 after the cells were co-exposed to1 µg/ml filipin and 100 µg/ml CB NPs.

CONCLUSIONCarbon black nanoparticles can be transferred into A549 cells by endocytosis, but caveolin-mediated endocytic pathway plays a minor role in this process.

Caveolin 1 ; physiology ; Cell Line ; Endocytosis ; Humans ; Nanoparticles ; Soot ; pharmacokinetics

Nanotoxicological research has shown toxicity of nanomaterials to be inversely related to particle size. However, the contribution of agglomeration to the toxicity of nanomaterials has not been sufficiently studied, although it is known that agglomeration is associated with increased nanomaterial size. In this study, we prepared aerosols of nano-sized carbon black by 2 different ways to verify the effects of agglomeration on the toxicity and deposition of nano-sized carbon black. The 2 methods of preparation included the carbon black dispersion method that facilitated clustering without sonication and the carbon black dispersion method involving sonication to achieve scattering and deagglomeration. Male Sprague-Dawley rats were exposed to carbon black aerosols 6 hr a day for 3 days or for 2 weeks. The median mass aerodynamic diameter of carbon black aerosols averaged 2.08 microm (for aerosol prepared without sonication; group N) and 1.79 microm (for aerosol prepared without sonication; group S). The average concentration of carbon black during the exposure period for group N and group S was 13.08 +/- 3.18 mg/m3 and 13.67 +/- 3.54 mg/m3, respectively, in the 3-day experiment. The average concentration during the 2-week experiment was 9.83 +/- 3.42 mg/m3 and 9.08 +/- 4.49 mg/m3 for group N and group S, respectively. The amount of carbon black deposition in the lungs was significantly higher in group S than in group N in both 3-day and 2-week experiments. The number of total cells, macrophages and polymorphonuclear leukocytes in the bronchoalveolar lavage (BAL) fluid, and the number of total white blood cells and neutrophils in the blood in the 2-week experiment were significantly higher in group S than in normal control. However, differences were not found in the inflammatory cytokine levels (IL-1beta, TNF-alpha, IL-6, etc.) and protein indicators of cell damage (albumin and lactate dehydrogenase) in the BAL fluid of both group N and group S as compared to the normal control. In conclusion, carbon black aerosol generated by sonication possesses smaller nanoparticles that are deposited to a greater extent in the lungs than is aerosol formulated without sonication. Additionally, rats were narrowly more affected when exposed to carbon black aerosol generated by sonication as compared to that produced without sonication.
Aerosols ; Animals ; Bronchoalveolar Lavage ; Carbon ; Humans ; Interleukin-6 ; Lactic Acid ; Leukocytes ; Lung ; Macrophages ; Male ; Nanoparticles ; Nanostructures ; Neutrophils ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Sonication ; Soot ; Tumor Necrosis Factor-alpha

Anthracosis is a black pigmentation of bronchial mucosa or lung parenchyma induced by inhaled soot. It is most commonly found in those persons who have worked as coal miners. Anthracosis is known to be induced when coal dust remains on the surface of the mucous membranes after it is inhaled through the bronchi, which can cause changes or atrophy of the upper airway and abnormality of the self-purification function and it can disrupt the defensive reaction of the respiratory organs. To our knowledge, there has been no report of recurrent paranasal sinusitis caused by anthracosis. Herein, with a review of the literature, we report a rare case of paranasal sinusitis caused by anthracosis in a 51-year-old man.
Anthracosis ; Atrophy ; Bronchi ; Coal ; Dust ; Humans ; Lung ; Mucous Membrane ; Pigmentation ; Sinusitis ; Soot

The biological activity of particles is largely dependent on their size in biological systems. Dispersion in the aqueous phase has been both a critical impediment to and a prerequisite for particle studies. Carbon black has been used as a surrogate to investigate the biological effects of carbonaceous particles. Here, biocompatible methods were established to disperse carbon black into ultrafine and fine particles which are generally distinguished by the small size of 100 nm. Carbon black with a distinct particle size, N330 and N990 were suspended in blood plasma, cell culture media, Krebs-Ringer's solution (KR), or physiological salt solution (PSS). Large clumps were observed in all dispersion preparations; however, sonication improved dispersion - averaged particle sizes for N330 and N990 were 85.0 +/- 42.9 and 112.4 +/- 67.9 nm, respectively, in plasma; the corresponding sizes in culture media were 84.8 +/- 38.4 and 164.1 +/- 77.8 nm. However, sonication was not enough to disperse N330 less than 100 nm in either KR or PSS. Application of Tween 80 along with sonication reduced the size of N330 to less than 100 nm, and dispersed N990 larger than 100 nm (73.6 +/- 28.8 and 80.1 +/- 30.0 nm for N330 and 349.5 +/- 161.8 and 399.8 +/- 181.1 nm for N990 in KR and PSS, respectively). In contrast, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) exhibited little effect. Electron microscopy confirmed the typical aciniform structure of the carbon arrays; however, zeta potential measurement failed to explain the dispersibility of carbon black. The methods established in this study could disperse carbon black into ultrafine and fine particles, and may serve as a useful model for the study of particle toxicity, particularly size-related effects.
1,2-Dipalmitoylphosphatidylcholine ; Carbon ; Cell Culture Techniques ; Culture Media ; Microscopy, Electron ; Particle Size ; Plasma ; Polysorbates ; Silicones ; Sonication ; Soot

OBJECTIVES: Recent studies have shown that nano-sized carbon black is more toxic than large respirable carbon black because of its higher surface area. However, it is not clear if carbon black made larger by agglomeration demonstrates decreased toxicity. The purpose of this study was to verify if agglomeration affects the toxicity of carbon black using three differently prepared nano-sized carbon black aerosols in nose-only inhalation chambers for 13 weeks. METHODS: Printex 90 was selected as a representative nano-sized carbon black. To generate aerosols of three different types of agglomerates, Printex 90 was dispersed in distilled water by three different methods: vortex, vortex+sonication, and vortex+sonication with dispersion in a stabilizer. Then, the three differently prepared solutions were aerosolized through venturi nozzles. Male Sprague-Dawley rats were exposed to Printex 90 aerosols in a nose-only exposure chamber for 6 h/d, 5 d/wk for 13 weeks at a concentration of approximately 9 mg/m3. RESULTS: Numbers of total cells in the bronchoalveolar lavage (BAL) fluid, macrophages, and polymorphonuclear leukocytes were increased and carbon black masses were clearly seen in BAL cells and lung tissues of rats exposed to Printex 90. However, few differences were found between the three differently agglomerated aerosols. In addition, there were no significant differences in other parameters, such as body weight, lung function or cytokine levels in BAL fluid following carbon black exposure. CONCLUSIONS: Only mild to moderate respiratory effects were found in rats exposed to nano-sized carbon black at 9 mg/m3 for 13 weeks. Agglomeration did not affect the toxicity of nano-sized carbon particles.
Aerosols ; Animals ; Body Weight ; Bronchoalveolar Lavage ; Carbon ; Humans ; Inhalation ; Lung ; Macrophages ; Male ; Neutrophils ; Rats ; Rats, Sprague-Dawley ; Soot ; Water