Pavel Coufalík

Plant-based dairy alternatives (PBDAs) have become essential components of the diet for individuals adhering to vegetarian, vegan, and flexitarian diets. In this study, levels of essential (Ca, P, Fe, Zn) and toxic (Cd, Pb) elements were assessed in commercially available and homemade PBDAs, including non-fortified plant-based milk, cheese, yogurt, cream alternatives, and tofu. The obtained data revealed a critical deficiency of calcium in non-fortified PBDAs, with a concerningly low Ca/P ratio. In the least favorable scenario modeled, consuming three servings of PBDAs would only provide a hypothetical daily calcium intake of 10 mg. The toxic elements lead and cadmium were generally very low, with the exception of homemade poppy seed milk, exhibiting an elevated cadmium level (0.052 ± 0.04 mg/kg). These findings highlight the importance of calcium source diversification in vegan diets, potentially through fortified plant-based foods or selected vegetables.

This study investigates the viability of Antarctic regolith and a lunar regolith simulant as prospective substrates for plant growth under extreme conditions. The study is focused on chemical composition of both substrates with a special respect to heavy metals. The Antarctic regolith samples were collected from James Ross Island, while lunar regolith simulant was developed in order to access the mobility and bioavailability of trace metals in these substrates, using a modified BCR (Community Bureau of Reference) sequential extraction procedure. Cadmium (Cd) and zinc (Zn) are among the significant mobile fractions, indicating their possible effects on the germination and growth of plants. Additionally, preliminary results from the study of the regolith samples at the area of Berry Hill mesa are presented, as well as the chemical and mineralogical affinity of the Antarctic regolith and the produced lunar simulant to lunar regolith. The findings suggest that such Antarctic environment can be used in astrobiological studies and future lunar habitation efforts to sustainably cultivate plants and perform other biological experiments on these extraterrestrial surfaces. This is an important concept in light of active programs like Artemis and Gateway.

The northern part of James Ross Island is the largest deglaciated area in the Antarctic Peninsula region with a unique ecosystem created during the Late Glacial. This research aims to evaluate the degree of contamination of the locality with toxic metals (As, Hg, Cd, and Pb) through bioindicators in the aquatic environment—colonies of cyanobacteria and algae. For this purpose, bottom lake sediments of Big Lachman Lake were studied for contents of Fe, As, Hg, Cd, Pb, Cr, Co, Ni, Cu, and Zn, as well as samples of cyanobacterial mat, in which Fe, As, Hg, Cd, and Pb were determined. Metal contents were determined by means of inductively coupled plasma optical emission spectrometry and atomic absorption spectrometry. The contents of metals in sediments did not differ from the usual values in the area of the Antarctic Peninsula. The bioaccumulation of metals in cyanobacterial mat was evaluated by calculating enrichment factors (the calculation to Fe as a reference element). According to this method, moderate pollution of Big Lachman Lake was confirmed for Hg and Cd. Graphical abstract

Titanium dioxide nanoparticles (TiO₂ NPs) are used in a wide range of applications. Although inhalation of NPs is one of the most important toxicologically relevant routes, experimental studies on potential harmful effects of TiO₂ NPs using a whole-body inhalation chamber model are rare. In this study, the profile of lymphocyte markers, functional immunoassays, and antioxidant defense markers were analyzed to evaluate the potential adverse effects of seven-week inhalation exposure to two different concentrations of TiO₂ NPs (0.00167 and 0.1308 mg TiO₂/m³) in mice. A dose-dependent effect of TiO₂ NPs on innate immunity was evident in the form of stimulated phagocytic activity of monocytes in low-dose mice and suppressed secretory function of monocytes (IL-18) in high-dose animals. The effect of TiO₂ NPs on adaptive immunity, manifested in the spleen by a decrease in the percentage of T-cells, a reduction in T-helper cells, and a dose-dependent decrease in lymphocyte cytokine production, may indicate immunosuppression in exposed mice. The dose-dependent increase in GSH concentration and GSH/GSSG ratio in whole blood demonstrated stimulated antioxidant defense against oxidative stress induced by TiO₂ NP exposure.

Due to widespread use in various industrial processes, the risk of inhalation exposure to lead oxide nanoparticles (PbO NPs) is growing and limited information evokes the need for assessment of hazard to human health. The aim of our work was to evaluate the effects of PbO NPs on blood, immune response, kidneys, intestine, and bones in mice exposed to 121.7 μg PbO per m³ continuously for 6 weeks. Immune assays showed significantly increased phagocytic activity of granulocytes, stimulated proliferative activity of spleen lymphocytes, a higher percentage of spleen CD3⁻CD335⁺ and thymus CD3⁺CD8⁺ cells, and moderately increased proinflammatory cytokines in the PbO NP-exposed group. Analysis of blood antioxidant defense revealed a significantly lower GSH concentration and a decrease in the GSH/GSSG ratio due to PbO NP inhalation. Markers of oxidative stress (AGEs and AOPP) in the intestine, but not in the kidneys, were significantly increased. In the kidney of PbO NP-treated mice, we found elevated expression of proinflammatory factors (TNF-α, IL-6, iNOS, and Xdh) and IL-10 and upregulated expression of F4/80 (mature macrophage marker). Moreover, increased expression of pro-fibrotic markers (TGFβ2, type I and type III collagen) was detected. Massive accumulation of Pb together with a significant decrease of Ca, P, and Mg concentrations in the tibia were accompanied by a nonsignificant increase in bone mineral density. Our findings indicate adverse effects of sub-chronic inhalation of PbO NPs, manifested as undesired stimulation of the immune system, oxidative stress in the intestine, inflammation, and pro-fibrotic response in the kidneys and massive accumulation of Pb in bones.

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 (TGFβ1), interleukin 6(IL-6), IL-1α and IL-1β , 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.

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.

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).

The use of alternative fuels or biofuel blends could be a way to reduce the environmental burden of increasing traffic. The aim of this study was to compare emissions from conventional fuels and alternative biofuels for diesel and spark-ignition engines under comparable conditions, i.e., using the World Harmonized Transient Cycle for a heavy-duty diesel engine and the Artemis CADC driving cycle for automobiles powered by gasoline and alternative fuels. Total contents of Ba, Ce, Cd, Cr, Cu, Fe, Mn, Ni, Pb, V, and Zn were determined in emissions, fuels, and lubricating oils. In addition, the bioaccessibility of metals in emissions was also assessed by extraction in water and in simulated lung fluid (Gamble’s solution). Total particulate mass emissions, expressed per kilogram of fuel, and total contents of metals were higher for the diesel engine than for spark-ignition engines. The highest metal contents in emissions from diesel and gasoline fuels were found for Fe, Zn, and Cu. Fe and Cu in emissions from diesel and spark-ignition engines declined with the addition of bio-components in fuels. However, there was no significant decrease in the contents of other metals in emissions from biofuels. The highest degrees of bioaccessibility were observed for Ba, Zn, Cd, and V in emissions from diesel and biodiesel (according to their solubility in water). On the basis of this study, the use of biodiesel (especially methylesters of rapeseed oil) can be recommended to reduce the total mass of particulate and metal emissions from diesel engines.