Carbon-based nanomaterials (C-BNM) have recently attracted an increased attention as the materials with potential applications in industry and medicine. Bioresistance and proinflammatory potential of C-BNM is the main obstacle for their medicinal application which was documented in vivo and in vitro. However, there are still limited data especially on graphene derivatives such as graphene platelets (GP). In this work, we compared multi-walled carbon nanotubes (MWCNT) and two different types of pristine GP in their potential to activate inflammasome NLRP3 (The nod-like receptor family pyrin domain containing 3) in vitro. Our study is focused on exposure of THP-1/THP1-null cells and peripheral blood monocytes to C-BNM as representative models of canonical and alternative pathways, respectively. Although all nanomaterials were extensively accumulated in the cytoplasm, increasing doses of all C-BNM did not lead to cell death. We observed direct activation of NLRP3 via destabilization of lysosomes and release of cathepsin B into cytoplasm only in the case of MWCNTs. Direct activation of NLRP3 by both GP was statistically insignificant but could be induced by synergic action with muramyl dipeptide (MDP), as a representative molecule of the family of pathogen-associated molecular patterns (PAMPs). This study demonstrates a possible proinflammatory potential of GP and MWCNT acting through NLRP3 activation.

To find the molecular components with the highest underlying probability of sensitization in patients suffering from atopic dermatitis and in subgroup of patients with allergic rhinitis, asthma bronchiale and to determine whether there is some difference of sensitization in mild, moderate and severe form of AD, in patients suffering from asthma bronchiale and allergic rhinitis. The complete dermatological and allergological examination including the examination of the sensitization to molecular allergens with Multiplex ISAC testing was performed. Altogether 100 atopic dermatitis patients were examined-48 men, 52 women, the average age 40.9 years; the sensitization to molecular components was confirmed in 93% of patients. The highest observed sensitization rate was 61.0% to grass specific molecule Phl p 1, the second most frequent sensitization was 57.0% to Betulaceae-specific molecule Bet v 1. Frequently observed sensitizations were those to PR-10 proteins, NPC2 proteins family, Uteroglobin and Lipocalin. In severe form of atopic dermatitis sensitization to molecular components of NPC2 proteins family, Uteroglobin, Lipocalin and Aspergillus were recorded with a significant higher occurrence.

The evaluation of carbon-based nanomaterials’ (C-BNMs’) interactions with the immune system, notably their ability to cause inflammation, is a critical step in C-BNM health risk assessment. Particular attention should be given to those C-BNMs that do not cause direct cytotoxicity or inflammation on their own. However, the intracellular presence of these non-biodegradable nanomaterials could dysregulate additional cell functions. This is even more crucial in the case of phagocytes, which are the main mediators of defensive inflammation towards pathogens. Hence, our study was focused on multi-walled carbon nanotubes (MWCNTs) and two different types of graphene platelets (GPs) and whether their intracellular presence modulates a proinflammatory response from human primary monocytes towards common pathogens. Firstly, we confirmed that all tested C-BNMs caused neither direct cytotoxicity nor the release of tumour necrosis factor α (TNF-α), interleukin (IL)-6 or IL-10. However, such pre-exposed monocytes showed increased responsiveness to additional bacterial stimuli. In response to several types of bacteria, monocytes pre-treated with GP1 produced a significantly higher quantity of TNF-α, IL-6 and IL-10. Monocytes pre-treated with MWCNTs produced increased levels of IL-10. All the tested C-BNMs enhanced monocyte phagocytosis and accelerated their differentiation towards macrophages. This study confirms the immunomodulatory potential of C-BNMs.

Aim: The cluster analysis for the evaluation of the results in Multiplex ISAC test in atopic dermatitis patients. Method: The complete dermatological and allergological examination including the examination of the sensitisation to molecular components with Multiplex ISAC testing was performed. The cluster analysis of molecular components (silhouette value) was processed. Results and conclusion: Altogether 100 atopic dermatitis patients were examined -48 men, 52 women, the average age 40.9 years. We found 10 clusters with different numbers of molecular components. Fundamental position have the components Phl p 1 (Timothy), Bet v 1 (Birch), Alt a 1 (Alternaria) followed by molecular components of NPC2 family, cystein proteasa, tropomyosin, uteroglobin, lipocalin and PR-10 protein. Our results correspond to the association of molecular components into protein families according to their biochemical structure.

Progress in laboratory diagnostics of IgE-mediated allergies is being made through the use of component-resolved diagnosis. The aim of our study is to analyze the sensitization profile to allergen reagents in patients suffering from atopic dermatitis with the use of the ALEX 2–Allergy Explorer and especially to show the sensitization to molecular components of molds and yeast. The complete dermatological and allergological examination including the examination of the sensitization to allergen reagents with Allergy Explorer ALEX 2 testing was performed. The relation between the sensitization to molecular components of molds and yeast and the severity of atopic dermatitis, and the occurrence of bronchial asthma and allergic rhinitis was evaluated. Altogether, 100 atopic dermatitis patients were examined—48 men and 52 women, with an average age of 40.9 years. The sensitization to Mala s 6, Mala s 11, Sac c, Asp f 6, Cla h and Cla h 8 correlates to the severity of atopic dermatitis. The sensitization to Sac c, Alt a 6, Cla h, Cla h 8 was observed significantly more frequently in patients suffering from bronchial asthma to Mala s 6 in patients suffering from allergic rhinitis. In patients with severe form of atopic dermatitis (AD), a very high level of specific IgE was recorded to Mala s 11 (in 36%) and to Asp f 6 (in 12%).

Background and aim: Progress in laboratory diagnostics of IgE-mediated allergy is the use of component-resolved diagnosis. Our study analyses the results of specific IgE to 295 allergen reagents (117 allergenic extracts and 178 molecular components) in patients suffering from atopic dermatitis (AD) with the use of ALEX2 Allergy Explorer. Method: The complete dermatological and allergological examination, including the examination of the sensitization to molecular components with ALEX2 Allergy Explorer testing, was performed. The statistical analysis of results was performed with these methods: TURF (total unduplicated reach and frequency), best reach and frequency by group size, two-sided tests, Fisher’s exact test, and chi-square test (at an expected minimum frequency of at least 5). Results: Altogether, 100 atopic dermatitis patients were examined: 48 men, 52 women, the average age 40.9 years, min. age 14 years, max. age 67 years. The high and very high level of specific IgE was reached in 75.0% of patients to 18 molecular components: from PR-10 proteins (Aln g 1, Bet v 1, Cor a1.0103, Cor a1.0401, Fag s 1), lipocalin (Can f 1), NPC2 family (Der f 2, Der p 2), uteroglobin (Fel d 1), from Alternaria alternata (Alt a 1), Beta expansin (Lol p 1, Phl p 1), molecular components from Timothy, cultivated rye (Secc pollen) and peritrophin-like protein domain Der p 23. The high and very high level of specific IgE to other lipocalins (Fel d 7, Can f 4), to arginine kinase (Bla g 9, German cockroach), and to allergen extracts Art v (mugwort), and Cyn d (Bermuda grass) reached 52.0% of patients. The severity of AD is in significant relation to the sensitization to molecular components of storage mites (Gly d 2, Lep d 2—NPC2 family), lipocalins (Can f 1, Can f 2, Can f 4, and Can f 6), arginine kinase (Asp f 6, Bla g 9, Der p 20, Pen m 2), uteroglobin (Fel d 1, Ory c 3), Mn superoxide dismutase (Mala s 11), PR-10 proteins (Fag s 1, Mal d 1, Cor a 1.0401, Cor a 1.0103), molecular components of the peritrophin-like domain (Der p 21, Der p 23), and to Secc pollen. In the subgroup of patients suffering from bronchial asthma, the significant role play molecular components from house dust mites and storage mites (Lep d 2, Der p 2, Der f 2—NPC2 family), cysteine protease (Der p 1), peritrophin-like protein domain (Der p 21, Der p 23), enolase from Alternaria alternata (Alt a 6), and Beta expansin Phl p 1. Conclusion: The results of our study demonstrate the detailed profile of sensitization to allergens reagents (allergen extract and molecular components) in patients with atopic dermatitis. We show the significance of disturbed epidermal barrier, resulting in increased penetration of allergens. We confirmed the significant relationship between the severity of AD, the occurrence of bronchial asthma and allergic rhinitis, and high levels of specific IgE to allergen reagents. Our results may be important for regime measures and immunotherapy; Der p 23 shall be considered as an essential component for the diagnosis and specific immunotherapy of house dust mite allergy.

(1) Background: Graphene is a two-dimensional atomic structure with a wide range of uses, including for biomedical applications. However, knowledge of its hazards is still limited. This work brings new cytotoxic, cytostatic, genotoxic and immunotoxic data concerning the in vitro exposure of human cell line to two types of graphene platelets (GP). It also contributes to the formation of general conclusions about the health risks of GP exposure. (2) Methods: In vitro exposure of a THP-1 cell line to three concentrations of two GP over 40 h. The cytotoxic potential was assessed by the measurement of LDH and glutathione (ROS) and by a trypan blue exclusion assay (TBEA); the cytostatic and genotoxic potential were assessed by the cytokinesis-block micronucleus (CBMN) test; and the immunotoxic potential was assessed by the measurement of IL-6, IL-10 and TNF-α. (3) Results: We found a significant dose-dependent increase in DNA damage (CBMN). The lowest observed genotoxic effect levels (LOGEL) were 5 µg/mL (GP1) and 30 µg/mL (GP2). We found no significant leaking of LDH from cells, increase in dead cells (TBEA), induction of ROS, increased levels of cytostasis, or changes in IL-6, IL-10 and TNF-α levels. (4) Conclusions: The genotoxicity increased during the short-term in vitro exposure of THP-1 to two GP. No increase in cytotoxicity, immunotoxicity, or cytostasis was observed.

Backround and aim: To evaluate the sensitisation to molecular components in atopic dermatitis patients with the use of Multiplex ISAC testing. Method: The complete dermatological and allergological examination including the examination of the sensitisation to molecular components with Multiplex ISAC testing was performed. Results and conclusion: Eighty one patients were examined -40 men, 41 women, average age 41.7 years. The positivity was recorded most frequently to molecular components of timothy, birch, house dust mite, animals dander, bermuda grass, peach, apple, hazel pollen, kiwi, peanut and hazelnut, Alternaria and Aspergillus. Molecular components such as Equ c 1 (Horse), Alt a 6 (Alternaria), Fel d 1, Fel d 4 (Cat) and Can f 1, Can f 5 (Dog) are recorded with the significant higher occurrence simultaneously in patients suffering from severe form of AD, asthma bronchiale and allergic rhinitis; these molecular components play the most important role in the atopic march.