A water-soluble [60]fullerene-derivative stimulates chlorophyll accumulation and has no toxic effect on Chlamydomonas reinhardtii

Chlamydomonas reinhardtii (WT 2137) P. A. Dang. (Volvocales, Chlorophyceae) is a green microalgae serving as a suitable model in scientific research and a promising industrial biotechnology platform for production of biofuel, hydrogen and recombinant proteins. Fullerenes (C60) are allotropic carbon nanoparticles discovered in 1985 and used in biomedical studies since the early 1990s, when water solubilization methodologies were developed. Recently, surface-modified hydroxylated derivatives of fullerenes were proven to enhance algal growth and drought tolerance in plants. Here, a novel type of water-soluble [60]fullerene derivative with 12 glycine residues (GF) has been synthesized and tested for acute toxicity (up to 50 μg/ml) and as a potential biostimulant of algal growth. The effects of GF on pigment composition and growth rate of Chlamydomonas reinhardtii were systematically investigated. Our results suggest that GF was not toxic, and no negative change in the pigment content and no stress symptoms were observed. No changes in the photosynthetic parameters based on the fluorescence of chlorophyll a in Photosystem II (NPQ, Fv/Fm, Fv/F0, PI and RC/ABS) were observed. The GF had no effect on cell size and growth rate. At a concentration of 20 μg/ml, GF stimulated chlorophyll accumulation in 3-day-old cultures

Mapping of DNA methylationsensitive cellular processes in gingival and periodontal ligament fibroblasts in the context of periodontal tissue homeostasis

Interactions between gingival fibroblasts (GFs) and oral pathogens contribute to the chronicity of inflammation in periodontitis. Epigenetic changes in DNA methylation are involved in periodontitis pathogenesis, and recent studies indicate that DNA methyltransferase (DNMT) inhibitors may protect against epithelial barrier disruption and bone resorption. To assess the impact of DNMT inhibition on GFs, cells were cultured with decitabine (5-aza-2’-deoxycytidine, DAC) for 12 days to induce DNA hypomethylation. We observed several potentially detrimental effects of DAC on GF biological functions. First, extended treatment with DAC reduced GF proliferation and induced necrotic cell death. Second, DAC amplified Porphyromonas gingivalis- and cytokine-induced expression and secretion of the chemokine CCL20 and several matrix metalloproteinases (MMPs), including MMP1, MMP9, and MMP13. Similar pro-inflammatory effects of DAC were observed in periodontal ligament fibroblasts. Third, DAC upregulated intercellular adhesion molecule-1 (ICAM-1), which was associated with increased P. gingivalis adherence to GFs and may contribute to bacterial dissemination. Finally, analysis of DAC-induced genes identified by RNA sequencing revealed increased expression of CCL20, CCL5, CCL8, CCL13, TNF, IL1A, IL18, IL33, and CSF3, and showed that the most affected processes were related to immune and inflammatory responses. In contrast, the genes downregulated by DAC were associated with extracellular matrix and collagen fibril organization. Our observations demonstrate that studies of DNMT inhibitors provide important insights into the role of DNA methylation in cells involved in periodontitis pathogenesis. However, the therapeutic potential of hypomethylating agents in periodontal disease may be limited due to their cytotoxic effects on fibroblast populations and stimulation of pro-inflammatory pathways.</p

Analiza globalnego profilu metylacji DNA fibroblastów dziąsłowych zdrowych osób i pacjentów z przewlekłym zapaleniem przyzębia przy użyciu sekwencjonowania wodorosiarczynowego o zredukowanej reprezentacji (ang. reduced representation bisulphite sequencing)

Niniejsza praca dotyczy badania wzorców metylacji DNA w fibroblastach dziąsłowych (GFs) uzyskanych od osób zdrowych (GFHD) i pacjentów z rozpoznanym przewlekłym zapaleniem przyzębia (GFPD). GFs odgrywają kluczową rolę w rozwoju chorób przyzębia i wykazują unikalne cechy jako „nieklasyczne” komórki wrodzonej odpowiedzi immunologicznej. Wcześniejsze badania koncentrowały się głównie na identyfikacji wzorców metylacji w całych tkankach, takich jak dziąsoa, krew i błona śluzowa policzków, uzyskanych od pacjentów z diagnozowaną paradontozą. Jednak status metylacji DNA poszczególnych populacji komórek, zwłaszcza GFs, pozostaje w dużej mierze niezbadany. W tej pracy zbadano stan metylacji różnych regionów genomowych i ogólny genom w GFs. Wyniki ujawniają różnice w profilach metylacji między dwiema grupami, nie tylko na poziomie genomu, ale także w obrębie określonych regionów, w tym regionów 5' nie podlegających translacji (z ang. 5′ untranslated region, 5'UTR), regionów 3' nie podlegających translacji (z ang. 3′ untranslated region, 3'UTR), długiego niekodującego RNA (lncRNA) i regionów intronowych. Dalsza analiza skupiła się na identyfikacji konkretnych regionów DNA o największej różnicy metylacji między dwiema badanymi grupami. Wbrew oczekiwaniom zidentyfikowane regiony były w większości niezwiązane z genami biorącymi udział w odpowiedzi immunologicznej. Zamiast tego wiązano je z różnymi funkcjami w różnych tkankach, od regulacji tworzenia tkanki zębowej do chorób nowotworowych i rozwóju neuronów. Dodatkowo analiza wzbogacenia funkcjonalnego (z ang. gene set enrichment analysis) ujawniła zmiany metylacji w regionach genów związanych między innymi ze szlakami sygnalizacji stanu zapalnego, aktywnością receptorów sprzężonych z białkiem G (ang. G-protein coupled receptors) oraz aktywnością czynników transkrypcyjnych. Głębsze poznanie i zrozumienie zmian w profilach metylacji DNA może przyczynić się do opracowania innowacyjnych markerów diagnostycznych i strategii terapeutycznych dla przewlekłego zapalenia przyzębia. Jednak konieczne są dalsze badania w celu potwierdzenia niniejszych wyników oraz wyjaśnienia dokładnych mechanizmów i funkcjonalnych konsekwencji związanych z zaobserwowanymi wzorcami metylacji.This study investigates the DNA methylation patterns present in gingival fibroblasts (GFs) obtained from healthy individuals (GFHD) and patients diagnosed with periodontitis (GFPD). GFs play a pivotal role in the development of periodontal diseases and exhibit unique characteristics as "non-classical" innate immune response cells. Previous research has predominantly focused on identifying methylation patterns in whole tissues, such as gingiva, blood, and buccal mucosa, obtained from periodontal patients. However, the DNA methylation status of individual cells, particularly GFs, has remained largely unexplored. In this study, the methylation status of various genomic regions and the overall genome in GFs was investigated. The results reveal differences in methylation profiles between the two groups, not only at the genome level but also within specific regions, including 5' untranslated regions (5'UTR), 3' untranslated regions (3'UTR), long non-coding RNA (lncRNA), and intronic regions. Further analysis focused on the identification of specific DNA regions with the highest difference between the two studied groups. Contrary to expectations, identified regions were mostly unrelated to genes involved in the immune response. Instead, they were associated with diverse functions in various tissues, ranging from dental tissue formation to cancer and neuronal development. Additionally, functional enrichment analysis uncovered methylation changes in regions of genes connected besides others to the inflammatory signalling pathways, G-coupled receptors activity and transcription factors activity. Gaining a comprehensive understanding of these alterations in DNA methylation profiles may contribute to the development of innovative diagnostic markers and therapeutic strategies for periodontitis. However, further research is necessary to confirm these findings and elucidate the exact mechanisms and functional consequences underlying these observed methylation patterns

Support for the Study of Geometric Transformations in the School Course by Means of Dynamic Mathematics Software

У статті проаналізовано комп’ютерний інструментарій програм динамічної математики GRAN2d, DG, Живая геометрия, Математический конструктор, GeoGebra у контексті підтримки вивчення геометричних перетворень площини. Виділено шляхи використання програм динамічної математики при вивченні теми «Геометричні перетворення на площині» з огляду на доцільність, раціональність та педагогічну виваженість їх використання.The article analyzes the computer tools of dynamic mathematics software GRAN2d, DG, The Geometer’s Sketchpad, MathKit, GeoGebra in the context of supporting the study of geometric plane transformations. The ways of using dynamic mathematics software in the study of the topic "Geometric transformations on a plane" are highlighted, considering the expediency, rationality and pedagogical weight of their use

Mapping of DNA methylation-sensitive cellular processes in gingival and periodontal ligament fibroblasts in the context of periodontal tissue homeostasis

Interactions between gingival fibroblasts (GFs) and oral pathogens contribute to the chronicity of inflammation in periodontitis. Epigenetic changes in DNA methylation are involved in periodontitis pathogenesis, and recent studies indicate that DNA methyltransferase (DNMT) inhibitors may protect against epithelial barrier disruption and bone resorption. To assess the impact of DNMT inhibition on GFs, cells were cultured with decitabine (5-aza-2’-deoxycytidine, DAC) for 12 days to induce DNA hypomethylation. We observed several potentially detrimental effects of DAC on GF biological functions. First, extended treatment with DAC reduced GF proliferation and induced necrotic cell death. Second, DAC amplified Porphyromonas gingivalis- and cytokine-induced expression and secretion of the chemokine CCL20 and several matrix metalloproteinases (MMPs), including MMP1, MMP9, and MMP13. Similar pro-inflammatory effects of DAC were observed in periodontal ligament fibroblasts. Third, DAC upregulated intercellular adhesion molecule-1 (ICAM-1), which was associated with increased P. gingivalis adherence to GFs and may contribute to bacterial dissemination. Finally, analysis of DAC-induced genes identified by RNA sequencing revealed increased expression of CCL20, CCL5, CCL8, CCL13, TNF, IL1A, IL18, IL33, and CSF3, and showed that the most affected processes were related to immune and inflammatory responses. In contrast, the genes downregulated by DAC were associated with extracellular matrix and collagen fibril organization. Our observations demonstrate that studies of DNMT inhibitors provide important insights into the role of DNA methylation in cells involved in periodontitis pathogenesis. However, the therapeutic potential of hypomethylating agents in periodontal disease may be limited due to their cytotoxic effects on fibroblast populations and stimulation of pro-inflammatory pathways

Mapping of DNA methylation-sensitive cellular processes in gingival and periodontal ligament fibroblasts in the context of periodontal tissue homeostasis

Interactions between gingival fibroblasts (GFs) and oral pathogens contribute to the chronicity of inflammation in periodontitis. Epigenetic changes in DNA methylation are involved in periodontitis pathogenesis, and recent studies indicate that DNA methyltransferase (DNMT) inhibitors may protect against epithelial barrier disruption and bone resorption. To assess the impact of DNMT inhibition on GFs, cells were cultured with decitabine (5-aza-2’-deoxycytidine, DAC) for 12 days to induce DNA hypomethylation. We observed several potentially detrimental effects of DAC on GF biological functions. First, extended treatment with DAC reduced GF proliferation and induced necrotic cell death. Second, DAC amplified Porphyromonas gingivalis- and cytokine-induced expression and secretion of the chemokine CCL20 and several matrix metalloproteinases (MMPs), including MMP1, MMP9, and MMP13. Similar pro-inflammatory effects of DAC were observed in periodontal ligament fibroblasts. Third, DAC upregulated intercellular adhesion molecule-1 (ICAM-1), which was associated with increased P. gingivalis adherence to GFs and may contribute to bacterial dissemination. Finally, analysis of DAC-induced genes identified by RNA sequencing revealed increased expression of CCL20, CCL5, CCL8, CCL13, TNF, IL1A, IL18, IL33, and CSF3, and showed that the most affected processes were related to immune and inflammatory responses. In contrast, the genes downregulated by DAC were associated with extracellular matrix and collagen fibril organization. Our observations demonstrate that studies of DNMT inhibitors provide important insights into the role of DNA methylation in cells involved in periodontitis pathogenesis. However, the therapeutic potential of hypomethylating agents in periodontal disease may be limited due to their cytotoxic effects on fibroblast populations and stimulation of pro-inflammatory pathways