Macrophage-mediated tissue response evoked by subchronic inhalation of lead oxide nanoparticles is associated with the alteration of phospholipases C and cholesterol transporters.

BACKGROUND: Inhalation of lead oxide nanoparticles (PbO NPs), which are emitted to the environment by high-temperature technological processes, heavily impairs target organs. These nanoparticles pass through the lung barrier and are distributed via the blood into secondary target organs, where they cause numerous pathological alterations. Here, we studied in detail, macrophages as specialized cells involved in the innate and adaptive immune response in selected target organs to unravel their potential involvement in reaction to subchronic PbO NP inhalation. In this context, we also tackled possible alterations in lipid uptake in the lungs and liver, which is usually associated with foam macrophage formation. RESULTS: The histopathological analysis of PbO NP exposed lung revealed serious chronic inflammation of lung tissues. The number of total and foam macrophages was significantly increased in lung, and they contained numerous cholesterol crystals. PbO NP inhalation induced changes in expression of phospholipases C (PLC) as enzymes linked to macrophage-mediated inflammation in lungs. In the liver, the subchronic inhalation of PbO NPs caused predominantly hyperemia, microsteatosis or remodeling of the liver parenchyma, and the number of liver macrophages also significantly was increased. The gene and protein expression of a cholesterol transporter CD36, which is associated with lipid metabolism, was altered in the liver. The amount of selected cholesteryl esters (CE 16:0, CE 18:1, CE 20:4, CE 22:6) in liver tissue was decreased after subchronic PbO NP inhalation, while total and free cholesterol in liver tissue was slightly increased. Gene and protein expression of phospholipase PLCß1 and receptor CD36 in human hepatocytes were affected also in in vitro experiments after acute PbO NP exposure. No microscopic or serious functional kidney alterations were detected after subchronic PbO NP exposure and CD68 positive cells were present in the physiological mode in its interstitial tissues. CONCLUSION: Our study revealed the association of increased cholesterol and lipid storage in targeted tissues with the alteration of scavenger receptors and phospholipases C after subchronic inhalation of PbO NPs and yet uncovered processes, which can contribute to steatosis in liver after metal nanoparticles exposure.

A simple dilute-and-shoot procedure for the determination of platinum in human pleural effusions using HR-CS GF-AAS.

BACKGROUND: High-resolution continuum source AAS is an emerging technique for the determination of trace elements in clinical analysis. We aimed to develop a method for the direct determination of platinum (Pt) in pleural effusions that could deepen the understanding of the dynamics of intrapleural Pt concentration during cytostatic therapy. MATERIALS AND METHODS: Samples were collected by thoracic drainage from five patients with lung cancer undergoing platinum based chemotherapy. A simple dilute-and-shoot method for Pt determination in the pleural effusions was developed. Ashing of the sample in an oxygen flow in a graphite tube allowed for direct analysis without prior mineralization. The trueness of the method was verified using an independent technique (ICP-MS). As platinum derivatives are only active in its free form (not bound to proteins), Pt in samples was further divided into free and protein-bound forms by means of ultrafiltration. RESULTS: Using the proposed method, Pt contents (free and total) were determined in samples collected at different times after the intravenous application of the Pt derivative. The concentration of total Pt reached values of up to 5,000 µg/L and different patterns of its dynamics in intrapleural fluid were observed. CONCLUSIONS: The developed method enables the fast and simple determination of Pt in biological fluids. It may be applied on a large scale to improve the understanding of Pt dynamics during cytostatic therapy, and also to determine the optimal timing of both thoracic drainage and administration of systemic chemotherapy.

A Clearance Period after Soluble Lead Nanoparticle Inhalation Did Not Ameliorate the Negative Effects on Target Tissues Due to Decreased Immune Response.

The inhalation of metal (including lead) nanoparticles poses a real health issue to people and animals living in polluted and/or industrial areas. In this study, we exposed mice to lead(II) nitrate nanoparticles [Pb(NO3)2 NPs], which represent a highly soluble form of lead, by inhalation. We aimed to uncover the effects of their exposure on individual target organs and to reveal potential variability in the lead clearance. We examined (i) lead biodistribution in target organs using laser ablation and inductively coupled plasma mass spectrometry (LA-ICP-MS) and atomic absorption spectrometry (AAS), (ii) lead effect on histopathological changes and immune cells response in secondary target organs and (iii) the clearance ability of target organs. In the lungs and liver, Pb(NO3)2 NP inhalation induced serious structural changes and their damage was present even after a 5-week clearance period despite the lead having been almost completely eliminated from the tissues. The numbers of macrophages significantly decreased after 11-week Pb(NO3)2 NP inhalation; conversely, abundance of alpha-smooth muscle actin (α-SMA)-positive cells, which are responsible for augmented collagen production, increased in both tissues. Moreover, the expression of nuclear factor κB (NF-κB) and selected cytokines, such as tumor necrosis factor alpha (TNFα), transforming growth factor beta 1 (TGFß1), interleukin 6(IL-6), IL-1α and IL-1ß , displayed a tissue-specific response to lead exposure. In summary, diminished inflammatory response in tissues after Pb(NO3)2 NPs inhalation was associated with prolonged negative effect of lead on tissues, as demonstrated by sustained pathological changes in target organs, even after long clearance period.

Determination of cadmium in lichens by solid sampling graphite furnace atomic absorption spectrometry (SS-GF-AAS).

The determination of trace metal contents directly from a solid sample is a trend in modern atomic spectrometry. The aim of this study was to develop an analytical method for the routine determination of Cd in lichens using solid sampling graphite furnace atomic absorption spectrometry (SS-GF-AAS). For the determination of Cd, the temperature program of the graphite furnace was optimized using a mixed matrix modifier (Pd + Mg (NO3)2 + Triton X-100). The limit of detection and the limit of quantification were 0.9 µg/kg and 3 µg/kg, respectively. The analytical method for Cd determination in the plant matrix was verified by the analysis of certified reference materials of lichens, seaweed, and rye grass. The developed procedure was applied to the study of Cd distribution in thalli of Usnea antarctica lichen from James Ross Island, Antarctica. The SS-GF-AAS analytical method is particularly suited for use in environmental studies and plant physiology (the microanalysis of anatomical structures).

Gene Expression and Epigenetic Changes in Mice Following Inhalation of Copper(II) Oxide Nanoparticles.

We investigated the transcriptomic response and epigenetic changes in the lungs of mice exposed to inhalation of copper(II) oxide nanoparticles (CuO NPs) (8 × 105 NPs/m3) for periods of 3 days, 2 weeks, 6 weeks, and 3 months. A whole genome transcriptome and miRNA analysis was performed using next generation sequencing. Global DNA methylation was assessed by ELISA. The inhalation resulted in the deregulation of mRNA transcripts: we detected 170, 590, 534, and 1551 differentially expressed transcripts after 3 days, 2 weeks, 6 weeks, and 3 months of inhalation, respectively. Biological processes and pathways affected by inhalation, differed between 3 days exposure (collagen formation) and longer treatments (immune response). Periods of two weeks exposure further induced apoptotic processes, 6 weeks of inhalation affected the cell cycle, and 3 months of treatment impacted the processes related to cell adhesion. The expression of miRNA was not affected by 3 days of inhalation. Prolonged exposure periods modified miRNA levels, although the numbers were relatively low (17, 18, and 38 miRNAs, for periods of 2 weeks, 6 weeks, and 3 months, respectively). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis based on miRNA-mRNA interactions, revealed the deregulation of processes implicated in the immune response and carcinogenesis. Global DNA methylation was not significantly affected in any of the exposure periods. In summary, the inhalation of CuO NPs impacted on both mRNA and miRNA expression. A significant transcriptomic response was already observed after 3 days of exposure. The affected biological processes and pathways indicated the negative impacts on the immune system and potential role in carcinogenesis.

Estimation of cadmium load from soybeans and soy-based foods for vegetarians.

Soybeans, as legumes, belong among food items that contribute most to the dietary intake of the heavy metal cadmium (Cd). The consumption of soy-based foods may contribute significantly to Cd intake in vegetarians as probably the most frequent consumers of these foods. In this research, the weekly dietary intake of Cd from soy-based foods was investigated in 119 participants (including vegetarians, vegans, and non-vegetarians) in the Czech Republic by means of a semi-quantitative food frequency questionnaire (SFFQ) with personal assistance and the measurement of body weight. Cd content in soy-based foods available on the market was determined by means of atomic absorption spectrometry after microwave digestion. Tofu, as the most frequently consumed food item in the vegetarian/vegan group, contributed most to the total dietary Cd intake. It contained 7.6 ± 0.1 µg Cd/kg, while the highest Cd content was observed in tempeh (18.1 ± 0.4 µg/kg). The highest mean dietary Cd intake per week from soy-based foods was 0.4 µg/kg b.w. and was found in the vegan group. The consumption of soy-based foods was the lowest in the non-vegetarian group as was the dietary intake of Cd, which was 0.04 µg/kg b.w. per week.

Simultaneous determination of cadmium and iron in different kinds of cereal flakes using high-resolution continuum source atomic absorption spectrometry.

A method for simultaneous determination of cadmium and iron in cereal flakes using high-resolution continuum source graphite furnace atomic absorption spectrometry is presented. Sample digest is introduced into the graphite furnace together with Pd/Mg(NO3)2 modifier. The primary absorption line of cadmium and adjacent secondary line of iron are used for the determination. Atomization is performed as a two-step process in order to meet ideal conditions for both elements. Interference produced by molecular absorption of PO molecular bands is suppressed by correction model using least squares background correction. Using the proposed method, levels of cadmium and iron were determined in different kinds of cereal flakes, where both elements are of great interest. Working range (0.01-2 µg L-1 for Cd and 10-500 µg L-1 for Fe) was suitable for the determination of analytes in samples. The method is fast, robust, and may be routinely used routinely in the analysis of foodstuffs.

Leukocyte telomere length is not affected by long-term occupational exposure to nano metal oxides.

The aim of this study was to ascertain whether long-term occupational exposure to nanoparticles would affect relative leukocyte telomere length (LrTL). We analysed occupational exposure to size-resolved aerosol particles, with special emphasis on nanoparticles at two workshops: i/ the production of nanocomposites containing metal oxides; ii/ laboratory to test experimental exposure of nano-CuO to rodents. Thirty five exposed researchers (age 39.5 ± 12.6 yr; exposure duration 6.0 ± 3.7 yr) and 43 controls (40.4 ± 10.5 yr) were examined. LrTL did not significantly (p=0.14) differ between the exposed researchers (0.92 ± 0.13) and controls (0.86 ± 0.15). In addition, no significant correlation (r=-0.22, p=0.22) was detected between the duration of occupational exposure and LrTL. The results remained non-significant after multiple adjustments for age, sex and smoking status. Our pilot results suggest that relative leukocyte telomere length is not affected by occupational exposure to nanoparticles.

The contents and distributions of cadmium, mercury, and lead in Usnea antarctica lichens from Solorina Valley, James Ross Island (Antarctica).

Lichens are efficient and cost-effective biomonitors of the environment. Their geographic distribution together with their slow growth rate enable investigation of the deposition patterns of various elements and substances. In this research, levels of cadmium, lead, and mercury in Usnea antarctica lichens in the area of James Ross Island, Antarctica, were investigated. The lichens were microwave-digested, and the metals were determined by means of atomic absorption spectrometry with graphite furnace and a direct mercury analyzer. Median total contents of Cd, Hg, and Pb were 0.04, 0.47, and 1.6 mg/kg in whole lichens, respectively. The bottom-up distributions of these metals in the fruticose lichen thalli were investigated, and it was revealed that the accumulation patterns for mercury and lead were opposite to that for cadmium. The probable reason for this phenomenon may lie in the inner structure of thalli. The total contents of metals were comparable with those published for other unpolluted areas of maritime Antarctica. However, this finding was not expected for mercury, since the sampling locality was close to an area with some of the highest mercury contents published for Antarctic lichens. In short, lichens proved their usability as biological monitors, even in harsh conditions. However, the findings emphasize the need to take into account the distributions of elements both in the environment and in the lichen itself.