Explore the vast range of titles available.

The authors describe a chemical approach for imaging deep into fixed brain tissue using Sca l e, a solution that renders biological samples transparent, but preserves fluorescent signals. This technique allows for imaging at unprecedented depth and at subcellular resolution, and makes three-dimensional reconstruction of neural networks possible without serial sectioning. Optical methods for viewing neuronal populations and projections in the intact mammalian brain are needed, but light scattering prevents imaging deep into brain structures. We imaged fixed brain tissue using Sca l e, an aqueous reagent that renders biological samples optically transparent but completely preserves fluorescent signals in the clarified structures. In Sca l e-treated mouse brain, neurons labeled with genetically encoded fluorescent proteins were visualized at an unprecedented depth in millimeter-scale networks and at subcellular resolution. The improved depth and scale of imaging permitted comprehensive three-dimensional reconstructions of cortical, callosal and hippocampal projections whose extent was limited only by the working distance of the objective lenses. In the intact neurogenic niche of the dentate gyrus, Sca l e allowed the quantitation of distances of neural stem cells to blood vessels. Our findings suggest that the Sca l e method will be useful for light microscopy–based connectomics of cellular networks in brain and other tissues.

Changes in HeLa cell morphology, membrane permeability, and viability caused by the presence of Triton X-100 (TX100), a nonionic surfactant, were studied by scanning electrochemical microscopy (SECM). No change in membrane permeability was found at concentrations of 0.15 mM or lower during an experimental period of 30 to 60 min. Permeability of the cell membrane to the otherwise impermeable, highly charged hydrophilic molecule ferrocyanide was seen starting at concentrations of TX100 of about 0.17 mM. This concentration level of TX100 did not affect cell viability. Based on a simulation model, the membrane permeability for ferrocyanide molecules passing though the live cell membrane was 6.5± 2.0 × 10⁻⁶ m/s. Cells underwent irreversible permeabilization of the membrane and structural collapse when the TX100 concentration reached the critical micelle concentration (CMC), in the range of 0.19 to 0.20 mM. The impermeability of ferrocyanide molecules in the absence of surfactant was also used to determine the height and diameter of a single living cell with the aid of the approach curve and probe scan methods in SECM.

Tissue-specific extracellular matrix plays an important role in promoting tissue regeneration and repair. We hypothesized that decellularized annular fibrosus matrix may be an appropriate scaffold for annular fibrosus tissue engineering. We aimed to determine the optimal decellularization method suitable for annular fibrosus. Annular fibrosus tissue was treated with 3 different protocols with Triton X-100, sodium dodecyl sulfate (SDS) and trypsin. After the decellularization process, we examined cell removal and preservation of the matrix components, microstructure and mechanical function with the treatments to determine which method is more efficient. All 3 protocols achieved decellularization; however, SDS or trypsin disturbed the structure of the annular fibrosus. All protocols maintained collagen content, but glycosaminoglycan content was lost to different degrees, with the highest content with TritonX-100 treatment. Furthermore, SDS decreased the tensile mechanical property of annular fibrosus as compared with the other 2 protocols. MTT assay revealed that the decellularized annular fibrosus was not cytotoxic. Annular fibrosus cells seeded into the scaffold showed good viability. The Triton X-100-treated annular fibrosus retained major extracellular matrix components after thorough cell removal and preserved the concentric lamellar structure and tensile mechanical properties. As well, it possessed favorable biocompatibility, so it may be a suitable candidate as a scaffold for annular fibrosus tissue engineering.

Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300gL−1), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142gL−1 after 5 days, which corresponded to 0.47gg−1 yield and productivity of 1.1gL−1h−1. Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol.

Triton X-100 (TX-100) is a widely used detergent to prevent viral contamination of manufactured biologicals and biopharmaceuticals, and acts by disrupting membrane-enveloped virus particles. However, environmental concerns about ecotoxic byproducts are leading to TX-100 phase out and there is an outstanding need to identify functionally equivalent detergents that can potentially replace TX-100. To date, a few detergent candidates have been identified based on viral inactivation studies, while direct mechanistic comparison of TX-100 and potential replacements from a biophysical interaction perspective is warranted. Herein, we employed a supported lipid bilayer (SLB) platform to comparatively evaluate the membrane-disruptive properties of TX-100 and a potential replacement, Simulsol SL 11W (SL-11W), and identified key mechanistic differences in terms of how the two detergents interact with phospholipid membranes. Quartz crystal microbalance-dissipation (QCM-D) measurements revealed that TX-100 was more potent and induced rapid, irreversible, and complete membrane solubilization, whereas SL-11W caused more gradual, reversible membrane budding and did not induce extensive membrane solubilization. The results further demonstrated that TX-100 and SL-11W both exhibit concentration-dependent interaction behaviors and were only active at or above their respective critical micelle concentration (CMC) values. Collectively, our findings demonstrate that TX-100 and SL-11W have distinct membrane-disruptive effects in terms of potency, mechanism of action, and interaction kinetics, and the SLB platform approach can support the development of biophysical assays to efficiently test potential TX-100 replacements.

Tissue-derived decellularized biomaterials are ideal for tissue engineering applications as they mimic the biochemical composition of the native tissue. These materials can be used as hydrogels for cell encapsulation and delivery. The decellularization process can alter the composition of the extracellular matrix (ECM) and thus influence the hydrogels characteristics. The aim of this study was to examine the impact of decellularization protocols in ECM-derived hydrogels obtained from porcine corneas. Porcine corneas were isolated and decellularized with SDS, Triton X-100 or by freeze-thaw cycles. All decellularization methods decreased DNA significantly when measured by PicoGreen and visually assessed by the absence of cell nuclei. Collagen and other ECM components were highly retained, as quantified by hydroxyproline content and sGAG, by histological analysis and by SDS-PAGE. Hydrogels obtained by freeze-thaw decellularization were the most transparent. The method of decellularization impacted gelation kinetics assessed by turbidimetric analysis. All hydrogels showed a fibrillary and porous structure determined by cryoSEM. Human corneal stromal cells were embedded in the hydrogels to assess cytotoxicity. SDS decellularization rendered cytotoxic hydrogels, while the other decellularization methods produced highly cytocompatible hydrogels. Freeze-thaw decellularization produced hydrogels with the overall best properties.

Surfactants are used for a variety of applications such as emulsifiers, solubilizers, or foaming agents. Their intensive production and use in pharmaceutical, cosmetic and agricultural products have resulted in their continuous discharge in the environment, especially via wastewaters. Surfactants have become a threat to living organisms as they interact with, and disrupt, cell membranes and macromolecules. Their effects have mainly been studied in aquatic species; however, terrestrial organisms are also threatened by these emerging contaminants. This study investigates the effects of two widely used nonionic surfactants, Tween-20 and Triton X-100, on key traits of larvae and adults of the fruit fly Drosophila melanogaster. We assessed the toxicity of the two surfactants on viability, development time, body size and food intake of the flies. The results revealed that both surfactants induced toxic effects on the drosophila flies leading to decreased viability, delayed development and lowered food consumption at the highest tested concentrations. Both surfactants proved to be toxic to flies, and, for all tested traits, Triton X-100 appeared more toxic than Tween-20. Our results might extend to other invertebrates. The widespread use of these substances, which then end up in the environment, should be regulated to mitigate their impacts on biodiversity and ecosystems.

To investigate the mechanism of Triton X-100 (TX-100) reducing the Ag + -resistance of Enterococcus faecalis ( E. faecalis ), and evaluate the antibacterial effect of TX-100 + Ag + against the induced Ag + -resistant E. faecalis (AR Ef ). The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of AgNO 3 against E. faecalis with/without TX-100 were determined to verify the enhanced antibacterial activity. Transmission electron microscopy (TEM) was used to observe the morphological changes of E. faecalis after treatment. The intra- and extracellular concentration of Ag + in treated E. faecalis was evaluated using inductively coupled plasma mass spectrometer (ICP-MS). The changes in cell membrane potential and integrity of treated E. faecalis were also observed using the flow cytometer. Moreover, AR Ef was induced through continuous exposure to sub-MIC of Ag + and the antibacterial effect of TX-100 + Ag + on AR Ef was further evaluated. The addition of 0.04% TX-100 showed maximal enhanced antibacterial effect of Ag + against E. faecalis . The TEM and ICP-MS results demonstrated that TX-100 could facilitate Ag + to enter E. faecalis through changing the membrane structure and integrity. Flow cytometry further showed the effect of TX-100 on membrane potential and permeability of E. faecalis . In addition, the enhanced antibacterial effect of TX-100 + Ag + was also confirmed on induced AR Ef . TX-100 can facilitate Ag + to enter E. faecalis through disrupting the membrane structure and changing the membrane potential and permeability, thus reducing the Ag + -resistance of E. faecalis and enhancing the antibacterial effect against either normal E. faecalis or induced AR Ef .

Soil washing is an effective approach to the removal of heavy metals from contaminated soil. In this study, the effects of the surfactants sodium dodecyl sulfate, Triton X-100, and non-ionic polyacrylamide (NPAM) on oxalic acid, tartaric acid, and citric acid used to remove zinc from contaminated soils were investigated. The Zn removal efficiencies of all washing solutions showed a logarithmic increase with acid concentrations from 0.5 to 10.0 g/L, while they decreased as pH increased from 4 to 9. Increasing the reaction time enhanced the effects of surfactants on Zn removal efficiencies by the acids during washing and significantly (P < 0.05) improved the removal under some mixed cases. Oxalic acid suffered antagonistic effects from the three surfactants and seriously damaged soil nutrients during the removal of soil Zn. Notably, the three surfactants caused synergistic effects on tartaric and citric acid during washing, with NPAM leading to an increase in Zn removal by 5.0 g/L citric acid of 10.60 % (P < 0.05) within 2 h. NPAM also alleviated the loss of cation exchange capacity of washed soils and obviously improved soil nitrogen concentrations. Overall, combining citric acid with NPAM offers a promising approach to the removal of zinc from contaminated soil.

Liver extracellular matrix (ECM)-based hydrogels have gained considerable interest as biomimetic 3D cell culture environments to investigate the mechanisms of liver pathology, metabolism, and toxicity. The preparation of current liver ECM hydrogels, however, is based on time-consuming thermal gelation and limits the control of mechanical properties. In this study, we used detergent-based protocols to produce decellularized porcine liver ECM, which in turn were solubilized and functionalized with methacrylic anhydride to generate photocrosslinkable methacrylated liver ECM (LivMA) hydrogels. Firstly, we explored the efficacy of two protocols to decellularize porcine liver tissue using varying combinations of commonly used chemical agents such as Triton X-100, Sodium Dodecyl Sulphate (SDS) and Ammonium hydroxide. Then, we demonstrated successful formation of stable, reproducible LivMA hydrogels from both the protocols by photocrosslinking. The LivMA hydrogels obtained from the two decellularization protocols showed distinct mechanical properties. The compressive modulus of the hydrogels was directly dependent on the hydrogel concentration, thereby demonstrating the tuneability of mechanical properties of these hydrogels. Immortalized Human Hepatocytes cells were encapsulated in the LivMA hydrogels and cytocompatibility of the hydrogels was demonstrated after one week of culture. In summary, the LivMA hydrogel system provides a simple, photocrosslinkable platform, which can potentially be used to simulate healthy versus damaged liver for liver disease research, drug studies and cancer metastasis modelling.