The cellular mechanisms involved in the development of silicosis have not been fully elucidated. This study aimed to examine influence of silica-induced lung injury on autophagy. Suspensions of crystalline silica particles were administered transnasally to C57BL/6j mice. Immunohistochemical examination for Fas and p62 protein expression was performed using lung tissue specimens. Two-dimensional and quantitative analysis of silica deposits in the lungs were performed in situ using lung tissue sections by an in-air microparticle induced X-ray emission (in-air micro-PIXE) analysis system, which was based on irrradiation of specimens with a proton ion microbeam. Quantitative analysis showed a significant increase of iron levels on silica particles (assessed as the ratio of Fe relative to Si) on day 56 compared with day 7 (p<0.05). Fas and p62 were expressed by histiocytes in granulomas on day 7, and the expressions persisted for day 56. Fas- and p62-expressing histiocytes were co-localized in granulomas with silica particles that showed an increase of iron levels on silica particles in mouse lungs. Iron complexed with silica induces apoptosis, and may lead to dysregulations of autophagy in histiocytes of granulomas, and these mechanisms may contribute to granuloma development and progression in silicosis.
Continuous irradiation effects on a thin-film diamond detector were investigated for the utilization of these films as a detector for heavy ion microbeams. Temporal signal degradation in the energy spectrum was frequently observed during the focused heavy ion microbeam irradiation. To measure the temporal response to the each ion incidents, focused heavy ion microbeam with different beam fluence rates were irradiated to a Single Crystal (SC)-CVD diamond film detector with thickness of 50 μm. The responses to each ion were continuously observed and characterized by ion beam-induced charge (IBIC) measurement system. Heavy ions with short penetration path in diamond generate the large difference in mean path of electrons and holes, which is inverted by changing bias polarity. Signal degradation condition was relied on the bias polarity under the irradiation of heavy ions with short penetration length in the diamond. The continuous observation of IBIC signals revealed that temporal degradation in pulse height of signals, so called polarization effects, seems to be mainly caused by the hole trapping in this diamond crystal.
Dental materials that are antimicrobial and acid-resistant can inhibit bacterial colonization and demineralization, thereby preventing caries. Zinc and copper are well-known for their antibacterial effect, as is nanostructured ZnO-CuO composite. Minerals such as fluorine and calcium, can remineralize and demineralize teeth. Therefore, we developed novel fluoride-containing ZnO-CuO (ZCF) nanocomposites; to the best of our knowledge, these are the first nanocomposites of this kind. The fluoride concentrations and antibacterial effects of the ZCF nanocomposites were evaluated. Nanocomposites comprising zinc and copper (ZC), and zinc, copper, and fluorine (ZCF), were prepared by a simple one-step homogeneous coprecipitation method at a low temperature (80°C), without the use of organic solvent or surfactant. The structure and composition of the ZC and ZCF nanocomposites were examined by scanning electron microscopy-energy-dispersive spectroscopy (SEM-EDS). Quantitative analysis of the mass concentration was performed by using ZAF correction methods. The fluorine content in nanocomposites was evaluated by using proton-induced gamma emission (PIGE) at the Takasaki Advanced Radiation Research Institute in Japan. By using 96-well microtiter plates, we analyzed the antibiotic susceptibility of ZC, ZCF, and the control buffer (phosphate-buffered saline) with Streptococcus mutans (ATCC 25175). The SEM images showed that ZC and ZCF nanocomposites were composed of 3D flower-like microstructures with diameters of approximately 1 μm. Environmental SEM-EDS analysis revealed that ZC contained 43.2% Cu, 55.1% Zn, 2.2% F, and 0.1% Cl, whereas ZCF contained 47.5% Cu, 40.5% Zn, 6.7% F, and 5.9% Cl. Analysis by PIGE showed that ZCF nanocomposite contained 2553.6 ± 199.2 ppm fluorine, whereas no fluoride was detected in ZC. The control buffer enabled bacterial growth to 4×10 7 ± 9×10 6 CFU/mL, whereas ZC allowed growth of 12 ± 8 CFU/mL, and ZCF showed no bacterial growth. Thus, we developed novel fluoride-containing ZnO-CuO nanocomposites, which exhibited antibacterial effects and have the potential for remineralization, thereby demonstrating their potential as multifunctional dental materials.
BackgroundPulmonary alveolar proteinosis (PAP) is a rare disease occurred by idiopathic (autoimmune) or secondary to particle inhalation. The in-air microparticle induced X-ray emission (in-air micro-PIXE) system performs elemental analysis of materials by irradiation with a proton microbeam, and allows visualization of the spatial distribution and quantitation of various elements with very low background noise. The aim of this study was to assess the secondary PAP due to inhalation of harmful particles by employing in-air micro-PIXE analysis for particles and intracellular iron in parafin-embedded lung tissue specimens obtained from a PAP patient comparing with normal lung tissue from a non-PAP patient. The iron inside alveolar macrophages was stained with Berlin blue, and its distribution was compared with that on micro-PIXE images.ResultsThe elements composing particles and their locations in the PAP specimens could be identified by in-air micro-PIXE analysis, with magnesium (Mg), aluminum (Al), silicon (Si), phosphorus (P), sulfur (S), scandium (Sc), potassium (K), calcium (Ca), titanium (Ti), chromium (Cr), copper (Cu), manganase (Mn), iron (Fe), and zinc (Zn) being detected. Si was the major component of the particles. Serial sections stained by Berlin blue revealed accumulation of sideromacrophages that had phagocytosed the particles. The intracellular iron content of alveolar macrophage from the surfactant-rich area in PAP was higher than normal lung tissue in control lung by both in-air micro-PIXE analysis and Berlin blue staining.ConclusionThe present study demonstrated the efficacy of in-air micro-PIXE for analyzing the distribution and composition of lung particles. The intracellular iron content of single cells was determined by simultaneous two-dimensional and elemental analysis of paraffin-embedded lung tissue sections. The results suggest that secondary PAP is associated with exposure to inhaled particles and accumulation of iron in alveolar macrophages.
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