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

Shotguns were developed as an alternative to single-missile rifle or handgun in order to enable gunner to spray a wide area with shot and thus kill birds and small animals too difficult to hit with single-missile weapons. As the popularity of shotguns grew, ammunition was developed for large game. The use of shotguns on humans has been markedly increasing in various manner of death. In forensic aspect, the shotgun wounds have characteristic findings due to their own characteristics and medical examiners may determine the range, distance, direction and kinds of gun. When the muzzle of the shotgun is placed tightly against the surface of the abdomen or thorax, the consequent wound of entrance will be single in number and circular in shape, and will have a diameter approximately equal to that of the bore of the weapon. However, when the wound is made over a site with underlying bone, especially in the scalp, the wound may have a different appearance, showing star shaped entrance due to tears from the sides of the wound and accompanying soot and burn effect. When the muzzle is held short to mid-range, considerable variation occurs in the appearance of the wound. We report two cases of shotgun wounds which showed typical findings of contact shots and distant shots.
Abdomen ; Animals ; Birds ; Burns ; Coroners and Medical Examiners ; Firearms ; Humans ; Scalp ; Soot ; Thorax ; Weapons ; Wounds and Injuries*

Shotguns were developed as an alternative to single-missile rifle or handgun in order to enable gunner to spray a wide area with shot and thus kill birds and small animals too difficult to hit with single-missile weapons. As the popularity of shotguns grew, ammunition was developed for large game. The use of shotguns on humans has been markedly increasing in various manner of death. In forensic aspect, the shotgun wounds have characteristic findings due to their own characteristics and medical examiners may determine the range, distance, direction and kinds of gun. When the muzzle of the shotgun is placed tightly against the surface of the abdomen or thorax, the consequent wound of entrance will be single in number and circular in shape, and will have a diameter approximately equal to that of the bore of the weapon. However, when the wound is made over a site with underlying bone, especially in the scalp, the wound may have a different appearance, showing star shaped entrance due to tears from the sides of the wound and accompanying soot and burn effect. When the muzzle is held short to mid-range, considerable variation occurs in the appearance of the wound. We report two cases of shotgun wounds which showed typical findings of contact shots and distant shots.
Abdomen ; Animals ; Birds ; Burns ; Coroners and Medical Examiners ; Firearms ; Humans ; Scalp ; Soot ; Thorax ; Weapons ; Wounds and Injuries*

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

Animals ; Carcinogenicity Tests ; Cytotoxicity Tests, Immunologic ; Humans ; Mutagenicity Tests ; Soot ; toxicity

OBJECTIVETo assess the genetic damage of human B lymphocyte cell line induced by 14 nm and 280 nm carbon black (CB) particles with micronucleus assay (CBMN), comet assay and hprt gene mutation test in vitro.

METHODSThe genetic damage of human B lymphocyte cells exposed to 14 nm and 280 nm CB particles at the doses of 0, 128, 256, 384 and 512 microg/ml for 24 h and 48 h was detected using above three genotoxic assays. Micronucleus (MN) assay, comet assay, hprt gene mutation test were used to detect the genetic damage of human B lymphocyte cells induced by CB. Micronucleus rate (MNR), micronucleated cell rate (MCR), nuclear buds (Buds), nucleoplasmic bridges (NPBs), nuclear division index (NDI) and numbers of apoptotic cells served as indexes of CBMN assay; the percentage of DNA in the tail (% tail DNA) and the olive tail moment (OTM) were used as DNA damage indicators of comet assay; the hprt gene mutation frequency (Mf-hprt) served as the index of hprt gene mutation test.

RESULTSThe % tail DNA, OTM in 14 nm CB group at the doses of 384 and 512 microg/ml for 48 h were 8.23% +/- 0.19%, 11.23% +/- 0.42% and 3.72 +/- 0.08, 4.90 +/- 0.18, respectively, which were significantly higher than those in control (5.10% +/- 0.08% and 2.22 +/- 0.03) (P < 0.01). The apoptotic cell rates in 14 nm CB group at the doses of 384 and 512 microg/ml for 48 h were 4.67 +/- 0.33 and 5.33 +/- 0.33, respectively, which were significantly higher than in control (0.00 +/- 0.00) (P < 0.05). The results of Mf-hprt were negative.

CONCLUSIONThe genetic damage of human B lymphocyte cells exposed to 14 nm CB particles for 48 h could be detected. But the similar effects didn't appear in 280 nm CB group.

B-Lymphocytes ; drug effects ; Cell Line ; Comet Assay ; DNA Damage ; drug effects ; Humans ; Micronucleus Tests ; Mutation Rate ; Soot ; toxicity

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

Deaths caused by grenades hardly occur except for during wartime, and can be seen as a particular incident that can be observed only within the military or by acts of terrorism. However, there has never been any data which tried to analyze the damage patterns through mock grenade explosions. So far, the processes leading to deaths were merely inferred by reconstructing the situation with limited intelligence after the incidents have already occurred. Therefore, the authors believe that having mock grenade explosions in order to compare and study the observances with those of the actual incident will be helpful for a more precise and objective medical jurisprudence, and thus have summarized the damage patterned obtained on 8 butchered pigs, that were 6~8 months old, about 100 cm in length and 70~100 kg in weight by photographing with radioactive rays and running autopsies after detonating grenades on them. In conclusion, if there was extensive loss of body parts, the grenade probably exploded when in contact with the lost body parts. If fractures were found the grenade is likely to have detonated contacting or within 25 cm of distance with the body, and if there were no lacerations or loss of body parts the grenade probably exploded at a distance of at least 50 cm. If soot was prevalent on the deceased's body, the grenade is likely to have exploded within a distance of 25 cm from the body at a free floating state, in which it was not in contact with the ground or the body. Lastly, if no soot was found, the grenade most probably detonated at a distance of 50 cm or more. The posture or the direction of the body cannot give precise clues of the victim's state before theexplosion, and a hasty presupposition could be very dangerous in case of contact explosions especially.
Autopsy ; Bombs ; Explosions ; Hand ; Homicide ; Human Body ; Humans ; Intelligence ; Jurisprudence ; Lacerations ; Military Personnel ; Posture ; Running ; Soot ; Suicide ; Swine ; Terrorism

OBJECTIVES: We sought to establish a novel method to generate nano-sized carbon black particles (nano-CBPs) with an average size smaller than 100 nm for examining the inhalation exposure risks of experimental rats. We also tested the effect of nano-CBPs on the pulmonary and circulatory systems. METHODS: We used chemical vapor deposition (CVD) without the addition of any additives to generate nano-CBPs with a particle size (electrical mobility diameter) of less than 100nm to examine the effects of inhalation exposure. Nano-CBPs were applied to a nose-only inhalation chamber system for studying the inhalation toxicity in rats. The effect on the lungs and circulatory system was determined according to the degree of inflammation as quantified by bronchoalveolar lavage fluid (BALF). The functional alteration of the hemostatic and vasomotor activities was measured by plasma coagulation, platelet activity, contraction and relaxation of blood vessels. RESULTS: Nano-CBPs were generated in the range of 83.3-87.9 nm. Rats were exposed for 4 hour/day, 5 days/week for 4 weeks to 4.2 x 10(6), 6.2 x 10(5), and 1.3 x 10(5) particles/cm3. Exposure of nano-CBPs by inhalation resulted in minimal pulmonary inflammation and did not appear to damage the lung tissue. In addition, there was no significant effect on blood functions, such as plasma coagulation and platelet aggregation, or on vasomotor function. CONCLUSION: We successfully generated nano-CBPs in the range of 83.3-87.9 nm at a maximum concentration of 4.2 x 10(6) particles/cm3 in a nose-only inhalation chamber system. This reliable method can be useful to investigate the biological and toxicological effects of inhalation exposure to nano-CBPs on experimental rats.
Animals ; Blood Platelets ; Bronchoalveolar Lavage Fluid ; Carbon ; Contracts ; Inflammation ; Inhalation ; Inhalation Exposure ; Lung ; Particle Size ; Plasma ; Platelet Aggregation ; Pneumonia ; Rats ; Relaxation ; Soot