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Frontiers in Cellular and Infection... 2024Enteroaggregative (EAEC) is a major cause of diarrhea worldwide. EAEC are highly adherent to cultured epithelial cells and make biofilms. Both adherence and biofilm...
Enteroaggregative (EAEC) is a major cause of diarrhea worldwide. EAEC are highly adherent to cultured epithelial cells and make biofilms. Both adherence and biofilm formation rely on the presence of aggregative adherence fimbriae (AAF). We compared biofilm formation from two EAEC strains of each of the five AAF types. We found that AAF type did not correlate with the level of biofilm produced. Because the composition of the EAEC biofilm has not been fully described, we stained EAEC biofilms to determine if they contained protein, carbohydrate glycoproteins, and/or eDNA and found that EAEC biofilms contained all three extracellular components. Next, we assessed the changes to the growing or mature EAEC biofilm mediated by treatment with proteinase K, DNase, or a carbohydrate cleavage agent to target the different components of the matrix. Growing biofilms treated with proteinase K had decreased biofilm staining for more than half of the strains tested. In contrast, although sodium metaperiodate only altered the biofilm in a quantitative way for two strains, images of biofilms treated with sodium metaperiodate showed that the EAEC were more spread out. Overall, we found variability in the response of the EAEC strains to the treatments, with no one treatment producing a biofilm change for all strains. Finally, once formed, mature EAEC biofilms were more resistant to treatment than biofilms grown in the presence of those same treatments.
Topics: Biofilms; Endopeptidase K; Escherichia coli; Deoxyribonucleases; Fimbriae, Bacterial; Bacterial Adhesion; Humans; Periodic Acid
PubMed: 38938878
DOI: 10.3389/fcimb.2024.1379206 -
BioImpacts : BI 2024Cell culture-based technologies are widely utilized in various domains such as drug evaluation, toxicity assessment, vaccine and biopharmaceutical development,... (Review)
Review
Cell culture-based technologies are widely utilized in various domains such as drug evaluation, toxicity assessment, vaccine and biopharmaceutical development, reproductive technology, and regenerative medicine. It has been demonstrated that pre-adsorption of extracellular matrix (ECM) proteins including collagen, laminin and fibronectin provide more degrees of support for cell adhesion. The purpose of cell imprinting is to imitate the natural topography of cell membranes by gels or polymers to create a reliable environment for the regulation of cell function. The results of recent studies show that cell imprinting is a tool to guide the behavior of cultured cells by controlling their adhesive interactions with surfaces. Therefore, in this review we aim to compare different cell cultures with the imprinting method and discuss different cell imprinting applications in regenerative medicine, personalized medicine, disease modeling, and cell therapy.
PubMed: 38938752
DOI: 10.34172/bi.2023.29945 -
Frontiers in Toxicology 2024Smoking cigarettes is a cause of serious diseases in smokers, including cardiovascular disease. Through a pathway of endothelial dysfunction, lipid infiltration,...
BACKGROUND
Smoking cigarettes is a cause of serious diseases in smokers, including cardiovascular disease. Through a pathway of endothelial dysfunction, lipid infiltration, macrophage recruitment and vascular remodeling, atherosclerosis is fundamental in the development of most cardiovascular diseases. There is an increasing number of next-generation products (NGP) which provide potentially reduced harm forms of nicotine delivery to adult smokers. This study aimed to optimise an cardiovascular model to assess such products. Human Coronary Artery Endothelial Cells (HCAECs) were cultured on an OrganoPlate2-lane chip (Mimetas BV) combined with THP-1 monocytes under flow conditions.
METHODS
An aqueous aerosol extract from the 1R6F reference cigarette was compared with two categories of NGP, (a heated tobacco product (HTP) and an electronic nicotine delivery system (ENDS)), to assess relative effects on select atherogenic endpoints (oxidative stress, monocyte adhesion, ICAM-1 expression, and inflammatory markers). Following exposure of THP-1 monocytes with the aqueous extracts, the resulting conditioned medium was then added to the HCAEC vessels.
RESULTS
1R6F was consistently the most potent test article, eliciting observed responses at 4x lower concentrations than applied for both the HTP and ENDS. The HTP was more potent than the ENDS product across all endpoints, however, all test articles increased monocyte adhesion. ICAM-1 did not appear to be a main driver for monocyte adhesion, however, this could be due to replicate variability. Upon comparison to an extract-only control exposure, THP-1-medium pre-conditioning was an important mediator of the responses observed.
CONCLUSION
In conclusion, the data suggests that the NGP extracts, containing primary aerosol chemical constituents exhibit a marked reduction in biological activity in the early key events associated with atherogenesis when compared to a cigarette, adding to the weight of evidence for the tobacco harm reduction potential of such products.
PubMed: 38938662
DOI: 10.3389/ftox.2024.1395670 -
Cell Surface (Amsterdam, Netherlands) Jun 2024Host recognition of the pathogen-associated molecular pattern (PAMP), β-1,3-glucan, plays a major role in antifungal immunity. β-1,3-glucan is an essential component...
Host recognition of the pathogen-associated molecular pattern (PAMP), β-1,3-glucan, plays a major role in antifungal immunity. β-1,3-glucan is an essential component of the inner cell wall of the opportunistic pathogen . Most β-1,3-glucan is shielded by the outer cell wall layer of mannan fibrils, but some can become exposed at the cell surface. In response to host signals such as lactate, shaves the exposed β-1,3-glucan from its cell surface, thereby reducing the ability of innate immune cells to recognise and kill the fungus. We have used sets of barcoded and mutants to compare the impacts of the secreted β-glucanases Xog1 and Eng1 upon and . Flow cytometry of Fc-dectin-1-stained strains revealed that Eng1 plays the greater role in lactate-induced β-1,3-glucan masking. Transmission electron microscopy and stress assays showed that neither Eng1 nor Xog1 are essential for cell wall maintenance, but the inactivation of either enzyme compromised fungal adhesion to gut and vaginal epithelial cells. Competitive barcode sequencing suggested that neither Eng1 nor Xog1 strongly influence fitness during systemic infection or vaginal colonisation in mice. However, the deletion of enhanced fitness during gut colonisation. We conclude that both Eng1 and Xog1 exert subtle effects on the cell surface that influence fungal adhesion to host cells and that affect fungal colonisation in certain host niches.
PubMed: 38938582
DOI: 10.1016/j.tcsw.2024.100128 -
The Japanese Dental Science Review Dec 2024This review examined the efficacy of surface treatments and adhesive monomers for enhancing zirconia-resin bond strength. A comprehensive literature search in PubMed,... (Review)
Review
This review examined the efficacy of surface treatments and adhesive monomers for enhancing zirconia-resin bond strength. A comprehensive literature search in PubMed, Embase, Web of Science, Scopus, and the Cochrane Library yielded relevant in vitro studies. Employing pairwise and Bayesian network meta-analyses, 77 articles meeting inclusion criteria were analyzed. Gas plasma was found to be ineffective, while treatments including air abrasion, silica coating, laser, selective infiltration etching, hot etching showed varied effectiveness. Air abrasion with finer particles (25-53 µm) showed higher immediate bond strength than larger particles (110-150 µm), with no significant difference post-aging. The Rocatec silica coating system outperformed the CoJet system in both immediate and long-term bond strength. Adhesives containing 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP) were superior to other acidic monomers. The application of 2-hydroxyethyl methacrylate and silane did not improve bonding performance. Notably, 91.2 % of bonds weakened after aging, but this effect was less pronounced with air abrasion or silica coating. The findings highlight the effectiveness of air abrasion, silica coating, selective infiltration etching, hot etching, and laser treatment in improving bond strength, with 10-MDP in bonding agents enhancing zirconia bonding efficacy.
PubMed: 38938474
DOI: 10.1016/j.jdsr.2024.05.004 -
Journal of Extracellular Biology Sep 2023Extracellular vesicles (EVs) recently emerged as important players in the pathophysiology of parasitic infections. While the protist parasite can produce EVs, their...
Extracellular vesicles (EVs) recently emerged as important players in the pathophysiology of parasitic infections. While the protist parasite can produce EVs, their role in giardiasis remains obscure. can disrupt gut microbiota biofilms and transform commensal bacteria into invasive pathobionts at sites devoid of colonizing trophozoites via unknown mechanisms. We hypothesized that EVs could modify gut bacterial behaviour via a novel mode of trans-kingdom communication. Our findings indicate that EVs exert bacteriostatic effects on HB101 and TW1, increasing their swimming motility. EVs also decreased the biofilm-forming ability of HB101 but not by TW1, supporting the hypothesis that these effects are, at least in part, bacteria-selective. HB101 and TW1 exhibited increased adhesion/invasion onto small intestine epithelial cells when exposed to EVs. EVs labelled with PKH67 revealed colocalization with HB101 and TW1 bacterial cells. Small RNA sequencing revealed a high abundance of ribosomal RNA (rRNA)- and transfer RNA (tRNA)-derived small RNAs, short-interfering RNAs (siRNAs) and micro-RNAs (miRNAs) within EVs. Proteomic analysis of EVs uncovered the presence of RNA chaperones and heat shock proteins that can facilitate the thermal stability of EVs and its sRNA cargo, as well as protein-modifying enzymes. In vitro, RNase heat-treatment assays showed that total RNAs in EVs, but not proteins, are responsible for modulating bacterial swimming motility and biofilm formation. small RNAs of EVs, but not proteins, were responsible for the increased bacterial adhesion to intestinal epithelial cells induced upon exposure to EVs. Together, the findings indicate that EVs contain a heat-stable, RNase-sensitive cargo that can trigger the development of pathobiont characteristics in Enterobacteria, depicting a novel trans-kingdom cross-talk in the gut.
PubMed: 38938375
DOI: 10.1002/jex2.109 -
Journal of Translational Medicine Jun 2024Over the last two decades, tumor-derived RNA expression signatures have been developed for the two most commonly diagnosed tumors worldwide, namely prostate and breast...
BACKGROUND
Over the last two decades, tumor-derived RNA expression signatures have been developed for the two most commonly diagnosed tumors worldwide, namely prostate and breast tumors, in order to improve both outcome prediction and treatment decision-making. In this context, molecular signatures gained by main components of the tumor microenvironment, such as cancer-associated fibroblasts (CAFs), have been explored as prognostic and therapeutic tools. Nevertheless, a deeper understanding of the significance of CAFs-related gene signatures in breast and prostate cancers still remains to be disclosed.
METHODS
RNA sequencing technology (RNA-seq) was employed to profile and compare the transcriptome of CAFs isolated from patients affected by breast and prostate tumors. The differentially expressed genes (DEGs) characterizing breast and prostate CAFs were intersected with data from public datasets derived from bulk RNA-seq profiles of breast and prostate tumor patients. Pathway enrichment analyses allowed us to appreciate the biological significance of the DEGs. K-means clustering was applied to construct CAFs-related gene signatures specific for breast and prostate cancer and to stratify independent cohorts of patients into high and low gene expression clusters. Kaplan-Meier survival curves and log-rank tests were employed to predict differences in the outcome parameters of the clusters of patients. Decision-tree analysis was used to validate the clustering results and boosting calculations were then employed to improve the results obtained by the decision-tree algorithm.
RESULTS
Data obtained in breast CAFs allowed us to assess a signature that includes 8 genes (ITGA11, THBS1, FN1, EMP1, ITGA2, FYN, SPP1, and EMP2) belonging to pro-metastatic signaling routes, such as the focal adhesion pathway. Survival analyses indicated that the cluster of breast cancer patients showing a high expression of the aforementioned genes displays worse clinical outcomes. Next, we identified a prostate CAFs-related signature that includes 11 genes (IL13RA2, GDF7, IL33, CXCL1, TNFRSF19, CXCL6, LIFR, CXCL5, IL7, TSLP, and TNFSF15) associated with immune responses. A low expression of these genes was predictive of poor survival rates in prostate cancer patients. The results obtained were significantly validated through a two-step approach, based on unsupervised (clustering) and supervised (classification) learning techniques, showing a high prediction accuracy (≥ 90%) in independent RNA-seq cohorts.
CONCLUSION
We identified a huge heterogeneity in the transcriptional profile of CAFs derived from breast and prostate tumors. Of note, the two novel CAFs-related gene signatures might be considered as reliable prognostic indicators and valuable biomarkers for a better management of breast and prostate cancer patients.
Topics: Humans; Prostatic Neoplasms; Male; Breast Neoplasms; Female; Cancer-Associated Fibroblasts; Gene Expression Regulation, Neoplastic; Prognosis; Transcriptome; Gene Expression Profiling; Cluster Analysis; Treatment Outcome; Middle Aged; Kaplan-Meier Estimate
PubMed: 38937754
DOI: 10.1186/s12967-024-05413-2 -
BMC Oral Health Jun 2024The aim of this study was to evaluate the adhesion of Candida glabrata, Candida albicans, Candida krusei, Candida parapsilosis and Candida tropicalis yeasts to...
BACKGROUND
The aim of this study was to evaluate the adhesion of Candida glabrata, Candida albicans, Candida krusei, Candida parapsilosis and Candida tropicalis yeasts to disk-shaped resin materials produced from resin which used in the production of surgical guide with 0, 45 and 90-degrees printing orientations by Liquid Crystal Display additive manufacturing technology.
METHODS
Disk-shaped specimens were printed with surgical guide resin using the Liquid Crystal Display production technique in 3 printing orientations (0, 45 and 90-degrees). Surface roughness and contact angle values were evaluated. Real-Time PCR analysis was performed to evaluate Candida adhesion (C. glabrata, C. albicans, C. krusei, C. parapsilosis and C. tropicalis) Field emission scanning electron microscope (FESEM) images of the materials were obtained.
RESULTS
Specimens oriented at 45-degrees demonstrated higher surface roughness (P < .05) and lower contact angle values than other groups. No significant difference was found in the adhesion of C. glabrata, C. albicans, and C. parapsilosis among specimens printed at 0, 45, and 90-degrees orientations (P > .05). A higher proportion of C. krusei and C. tropicalis was found in the specimens printed at orientation degrees of 45 = 90 < 0 with statistical significance. Analyzing the adhesion of all Candida species reveals no statistical disparity among the printing orientations.
CONCLUSIONS
The surface roughness, contact angle, and adhesion of certain Candida species are affected by printing orientations. Hence, careful consideration of the printing orientation is crucial for fabricating products with desirable properties. In 45-degree production, roughness increases due to the layered production forming steps, whereas in 0-degree production, certain Candida species exhibit high adhesion due to the formation of porous structures. Consequently, considering these factors, it is advisable to opt for production at 90-degrees, while also considering other anticipated characteristics.
Topics: Surface Properties; Candida; Microscopy, Electron, Scanning; Candida glabrata; Candida tropicalis; Candida parapsilosis; Humans; Candida albicans; Materials Testing; Cell Adhesion; Real-Time Polymerase Chain Reaction; Printing, Three-Dimensional; Resins, Synthetic; Wettability
PubMed: 38937749
DOI: 10.1186/s12903-024-04505-1 -
Scientific Reports Jun 2024The composite attachment loss during orthodontic clear aligner therapy is an adverse event that commonly happens in clinical practice and can affect the overall outcome...
The composite attachment loss during orthodontic clear aligner therapy is an adverse event that commonly happens in clinical practice and can affect the overall outcome and length of treatment. The aim of our research is to provide a basis for the further study of an innovative digital protocol and application method for orthodontic aligner attachments. Two 3D models were designed, one based on the proposed protocol and the other on the conventional method for aligner attachment application. Four attachment shapes were used to identify the maximum values for the von Mises equivalent stresses, the maximum displacements values and the areas in which these values were recorded through FEM analysis. The results of the mechanical simulation show lower values of von Mises stress recorded in the 3D printed attachments assemblies, independent of their shape, when simulated under the same boundary and load conditions. The trapezoidal prism shaped 3D printed model has a 3.7 times smaller displacement value (0.088 [mm]) compared to the adhesive resin model (0.326 [mm]). In conclusion, the proposed protocol for aligner attachments and the introduction of innovative materials is a promising method of solving conventional attachment problems in current orthodontic treatments.
Topics: Printing, Three-Dimensional; Finite Element Analysis; Humans; Stress, Mechanical; Tooth Movement Techniques; Orthodontic Appliance Design
PubMed: 38937650
DOI: 10.1038/s41598-024-65864-x -
Scientific Reports Jun 2024Both hypoxia and the complement lectin pathway (CLP) are involved in atherosclerosis and atherosclerosis-related stroke and acute myocardial infarction (AMI). We have...
Both hypoxia and the complement lectin pathway (CLP) are involved in atherosclerosis and atherosclerosis-related stroke and acute myocardial infarction (AMI). We have previously shown that mannose-binding lectin-associated serine protease-1 (MASP-1), the most abundant enzyme of CLP, induces an inflammatory phenotype of endothelial cells (ECs) by cleaving protease activated receptors (PARs). In the absence of data, we aimed to investigate whether hypoxia and MASP-1 interact at the level of ECs, to better understand their role in atherosclerosis-related diseases. Hypoxia attenuated the wound healing ability of ECs, increased ICAM-1 and decreased ICAM-2 expression and upregulated PAR2 gene expression. Hypoxia and MASP-1 increased GROα and IL-8 production, and endothelial permeability without potentiating each other's effects, whereas they cooperatively disrupted vascular network integrity, activated the Ca, CREB and NFκB signaling pathways, and upregulated the expression of E-selectin, a crucial adhesion molecule in neutrophil homing. VCAM-1 expression was not influenced either by hypoxia, or by MASP-1. In summary, hypoxia potentiates the effect of MASP-1 on ECs, at least partially by increasing PAR expression, resulting in interaction at several levels, which may altogether exacerbate stroke and AMI progression. Our findings suggest that MASP-1 is a potential drug target in the acute phase of atherosclerosis-related diseases.
Topics: Humans; Mannose-Binding Protein-Associated Serine Proteases; Atherosclerosis; Endothelial Cells; Signal Transduction; Cell Hypoxia; NF-kappa B; Receptor, PAR-2; Human Umbilical Vein Endothelial Cells; Intercellular Adhesion Molecule-1; E-Selectin; Interleukin-8
PubMed: 38937560
DOI: 10.1038/s41598-024-64479-6