Recently, generation of 3D spheroids from mesenchymal stromal cells (MSCs) for tissue engineering has been proposed, in order to recapitulate in vivo-like conditions, namely mild hypoxia and proper cell-cell interactions. The aim of the current study was to develop a novel simple approach to generate multicellular spheroids directly from monolayer cell culture of mesenchymal stromal cells. For this purpose, cell self-aggregation induced by the cyclo-RGDfK(TPP) peptide (5-100 µM) has been proposed. The cells were shown to form spheroids after adding the peptide to culture medium within 72h. The spheroids were stained with vital dye Calcein AM, and cell viability was confirmed by confocal laser scanning microscopy. The spheroids were able to reverse into single-cell culture after peptide removal, which provided cell invasion into the collagen hydrogel. The spheroid-based cultivation approach could be promising for tissue engineering.

Freshwater plastisphere structure and succession on diverse polymer surfaces: Linking plastisphere ecology and surface alterations in a 210-day field experiment.

A novel approach for achieving high enrichment of anammox and nitrogen removal rate in municipal wastewater treatment: A pure biofilm process.

Multi-omics reveal the mechanism of Plantaricin BM-1 in controlling yoghurt post-acidification by affecting the membrane and metabolism of Lactobacillus delbrueckii subsp. bulgaricus.

Enhancing synergistic partial nitrification-anammox coupling via step feed and granular sludge technology for optimized nitrogen removal in municipal wastewater treatment.

Valeriana wallichii-loaded tin oxide nanoparticle gel for targeted melanoma therapy: cytotoxicity, antioxidant activity, and cellular uptake evaluation.

A FRET-based ratiometric nanosensor for monitoring Acetyl-CoA levels in living cells.

Protocol for visualizing microglial lysosomal content by immunohistochemistry in the rodent brain.

High expression of IRF8 in AChR-specific cells regulates the function of B cells.

Astragaloside IV mitigates myocardial ischemia/reperfusion injury by modulating the 14-3-3η/GPX4 axis, reducing mitochondrial dysfunction, ferroptosis, and apoptosis.

To compare graft survival and alloimmune responses in murine endothelial keratoplasty (EK) versus penetrating keratoplasty (PK) and to elucidate the immunological mechanisms that underlie the differential graft outcomes. Allogeneic EK and PK were performed in BALB/c recipient mice using fully disparate C57BL/6 donors; syngeneic EK recipients served as controls. Graft clarity was monitored over 16 weeks by slitlamp biomicroscopy and scored using standardized opacity grading. Anterior segment optical coherence tomography (AS-OCT) was used to measure central corneal thickness. Graft survival was assessed using Kaplan-Meier analysis. Immunohistochemistry and confocal microscopy were performed to evaluate corneal endothelial cell (CEnC) integrity through ZO-1 staining. T-cell-mediated alloimmunity was assessed using intracellular IFN-γ staining (flow cytometry) and ELISPOT assays targeting both direct and indirect antigen presentation pathways. PK allografts exhibited significantly higher corneal opacity and lower survival (50%) than allogeneic EK grafts (71.4%, P < 0.0001). AS-OCT showed that corneal edema was highest in rejected PK grafts at 4 weeks and in rejected EK grafts at 16 weeks, with EK displaying a more gradual increase in thickness. Flow cytometry revealed significantly greater frequencies of IFN-γ + CD4 + T cells in PK recipients compared with EK recipients ( P < 0.001). ELISPOT assays demonstrated a more robust Th1 response in PK through both the direct and indirect sensitization pathways. Corneal endothelial cell (CEnC) density was significantly reduced in rejected EK and PK grafts compared with their respective accepted counterparts ( P < 0.01), whereas CEnC density was comparable between accepted EK and PK grafts. EK grafts exhibit higher graft survival rates and significantly reduced activation of host T-cell responses compared with PK grafts, which may be attributed to lower frequencies of graft-borne antigen presenting cells, thus resulting in a milder Th1-mediated immune response.

The purpose of this study was to characterize three recurrent, cancer-associated missense mutations in ErbB2, R143Q, R678Q, and V842I, located in the extracellular, juxtamembrane and kinase domains, respectively, to determine how single amino acid substitutions affect receptor organization and dynamics. Using confocal microscopy, Förster resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP), we assessed homo- and heteroassociation, lateral diffusion and tyrosine phosphorylation of ErbB2 cancer-associated variants either alone or co-expressed with EGFR. The only individual biophysical property that differentiated the mutation-activated ErbB2 variants from the wild-type was an accelerated diffusion in the absence of EGFR coexpression. Correlative analysis of the biophysical properties of ErbB2 revealed that ErbB2-activating mutations, including the cancer-associated R143Q, R678Q and V842I mutations, may promote a pre-dimerized receptor state associated with accelerated lateral mobility, but without full-scale activation implied by the lack of growth factor-independent tyrosine phosphorylation. The faster mobility of mutation-activated ErbB2 contrasted with the EGF-induced slowing down of its lateral diffusion. In summary, single amino acid substitutions across ErbB2 domains may modulate receptor dynamics, organization, and responsiveness.

To evaluate in vitro the antifungal activity, mechanism of action, drug association, and anti-biofilm effect of the essential oil of Lippia sidoides Cham. (EO-LS) against Candida albicans and Candida auris. EO-LS was extracted, and its chemical profile was determined by GC-MS. Assays were performed to determine the Minimum Inhibitory Concentration (MIC), Minimum Fungicidal Concentration (MFC), mechanism of action, drug interaction with nystatin and ketoconazole, time-kill kinetics, and biofilm inhibition by confocal microscopy. The compound exhibited fungicidal activity, with MIC and MFC values of 125 µg/mL against C. albicans and 62.5 µg/mL against C. auris. The exogenous ergosterol assay indicated that EO-LS exerts antifungal activity by interfering with fungal plasma membrane functions. The combination of EO-LS with ketoconazole demonstrated pharmacological synergism, while its combination with nystatin resulted in an antagonistic effect. EO-LS significantly inhibited fungal growth (p < 0.005) and promoted a marked reduction of mature C. auris biofilm at a concentration of 125 µg/mL (p < 0.0001), as confirmed by confocal microscopy imaging. The essential oil of Lippia sidoides exhibits antifungal activity against Candida spp., including C. auris, likely through an action on the cell membrane, and shows potential as an anti-biofilm agent with pharmacological synergism when combined with ketoconazole.

Zn2+-dependent modulation of the mitochondrial Ca2+ uniporter underlies resveratrol-mediated protection against myocardial ischemia-reperfusion injury.

The first pyrido[1,2-a]benzimidazole-fused porphyrazine photosensitizers with submicromolar photodynamic activity against cancer cells.

Protocol to study synapse density or volume-SynDOVE-in brain using confocal microscopy and Imaris three-dimensional surface rendering software.

Dry surface biofilm of Salmonella and Cronobacter sakazakii: a real concern for the low moisture food industry.

Microvilli are conserved actin-based protrusions that expand surface area and absorptive capacity in epithelial tissues. In humans, the intestinal tract grows 300 trillion microvilli per day, yet the molecular mechanisms defining where and when microvilli grow remain unclear. Using a combination of live-cell confocal and super-resolution microscopy to investigate native intestinal tissues and epithelial cell culture models, we found that microvilli grow from Arp2/3-generated branched actin networks that form on the surface of clathrin-coated pits. These transient networks are stabilized by cortactin and localize to the apical plasma membrane minutes before reorganizing into the linear core actin bundles that support microvilli. Moreover, subpixel precision localization of coated pit and microvilli markers revealed that F-actin asymmetrically localizes to one side of a nascent clathrin-coated pit, and that microvilli grow from the side with the highest F-actin density. These findings support a model where the asymmetric accumulation of F-actin and growing barbed ends on the surface of a clathrin-coated pit offers a filament source to support microvilli formation.

Chitosan enhances antimicrobial efficiency of ceftazidime against Burkholderia pseudomallei in an ex vivo skin model and cellular infections

Atractylodin ameliorates obesity-associated hepatic steatosis by regulating the PLIN2-ATGL/CPT1A axis-mediated lipid droplet-mitochondria interactions.