Triton X-100 Research Articles

Triton X-100-treated virus-based ELLA demonstrates discordant antigenic evolution of influenza B virus hemagglutinin and neuraminidase.

Neuraminidase (NA)-specific antibodies have been associated with protection against influenza and thus NA is considered a promising target for next-generation vaccines against influenza A (IAV) and B viruses (IBV). NA inhibition (NI) by antibodies is typically assessed using an enzyme-linked lectin assay (ELLA). However, ELLA can be confounded by anti-hemagglutinin (anti-HA) antibodies that block NA by steric hindrance (termed HA interference). Although strategies have been employed to overcome HA interference for IAV, similar approaches have not been assessed for IBV. We found that HA interference is common in ELLA using IBV, rendering the technique unreliable. Anti-HA antibodies were not completely depleted from sera by HA-expressing cell lines, and this approach was of limited utility. In contrast, we find that treatment of virions with Triton X-100, but not Tween-20 or ether, efficiently separates the HA and NA components and overcomes interference caused by anti-HA antibodies. We also characterize a panel of recombinant IBV NA proteins that further validated the results from Triton X-100-treated virus-based ELLA. Using these reagents and assays, we demonstrate discordant antigenic evolution between IBV NA and HA over the last 80 years. This optimized ELLA protocol will facilitate further in-depth serological surveys of IBV immunity as well as antigenic characterization of the IBV NA on a larger scale.IMPORTANCEInfluenza B viruses (IBVs) contribute to annual epidemics and may cause severe disease, especially in children. Consequently, several approaches are being explored to improve vaccine efficacy, including the addition of neuraminidase (NA). Antigen selection and assessment of serological responses will require a reliable serological assay to specifically quantify NA inhibition (NI). Although such assays have been assessed for influenza A viruses (IAVs), this has not been done of influenza B viruses. Our study identifies a readily applicable strategy to measure the inhibitory activity of neuraminidase-specific antibodies against influenza B virus without interference from anti-hemagglutinin (anti-HA) antibodies. This will aid broader serological assessment of influenza B virus-specific antibodies and antigenic characterization of the influenza B virus neuraminidase.

B-217 Effective Diagnostic Platform and Method to Differentiate the Infectious From Non-Infectious Viruses With Nucleic Acid Detection Workflow

Abstract Background While qPCR has demonstrated high sensitivity, specificity, and ability to deliver reliable quantitative data in clinical samples, such results do not differentiate between infectious and non-infectious viral particles. In view of the principle of the qPCR method itself, the amplification detects the nucleic acid (DNA/RNA) of the virus, not the virus itself. In many cases, it is still possible to detect DNA/RNA fragments from dead viruses in recovered patients, while they are not infectious anymore. This will lead to prolonged quarantine period or treatment for the patients, which could cause great psychological pressure to the patients and increased healthcare burden. Therefore, an effective detection method that can discriminate viable and inviable pathogens is of great significance. Methods SARS-CoV2 viral RNA, SARS-CoV-2 viruses, or clinical samples with SARS-CoV2 infection were diluted in Delta MTM (Molecular Transport Media) to the respective testing concentrations. The samples were incubated with intercalating dye (PMAxx 100nM) with or without Triton X-100 for 10 min. The mixture was then exposed to the photo-activation device for 5-15 min (@ temperature below 37° C). The samples were then subjected to the standard nucleic acid extraction & purification steps with Qiagen viral RNA extraction kit. The extracted RNA was subjected to amplification with SARS-CoV2 Resolute 2.0 & Fortunate 2.1 assays on BioRAD CFX96. Results The viability PCR (vPCR) efficiency was almost 100% for all SARS-CoV2 RNA input tested suggesting that without viral membrane protection, PMAxx dye effectively bound to the naked RNA after photoactivation process, and the bound RNA was not able to be amplified. The vPCR efficiency was further enhanced with addition of surfactant Triton X-100 suggesting the improved permeability of dye through the viral envelope facilitated the PMAxx binding to the nucleic acid. COVID19 positive clinical samples with viral load ≈ Ct 30 treated with or without PMAxx and photo-activation procedures showed distinctive ΔCt value ranging from 9.27±0.18 to 15.01±0.14. The Ct after dye treatment were all non-numeric (non-detectable) if cut-off Ct sets @ 42.0, indicating the complete inhibition after vPCR pretreatment. Since the clinical samples were undergone heat-inactivation pre-treatment, the result shown here indicating that all samples were non-infectious, which was consistent with the samples tested. Conclusions We developed an effective viability PCR detection method and proprietary vPCR device, which was specifically designed for effective dye photo-activation. In addition, the method takes less than 15 min and could be integrated into the existing standard nucleic acid extraction and amplification workflow. Comparing to the standard PCR test result, this method could provide the information not only on the positivity of the tested pathogen, but also its viability, which could be very essential information for public surveillance and diagnostic decisions for healthcare providers.

The Interaction Of Homodimer Styrylcyanine Dyes With Sodium Dodecyl Sulfate And Triton X-100.

Solubilization of the styrylcyanine dye Sbt ((E)-2-(4-(dimethylamino)styryl)-3-methylbenzo[d]thiazol-3-ium iodide) and its homodimers Dbt-5 and Dbt-10 in aqueous solution of sodium dodecyl sulfate and Triton X-100 has been studied by steady state and picosecond time-resolved fluorescence spectroscopy. At low concentration of sodium dodecyl sulfate in solution, between Sbt, Dbt-5 dyes molecules and surfactant ion pairs are formed followed by the formation non-luminescent H-aggregates. The nature of the interaction between molecules of dyes and surfactants has been revealed. The binding constants Ks of the dyes to the surfactants, free energy changes (ΔG0), the number of dye molecules (n) included in a single micelle and photophysical parameters have been determined. The degree of solubilization of dyes in micellar solution of Triton X-100 is higher as compared to sodium dodecyl sulfate and depends on the molecular weight and size of both dye molecules and micelles.

Permeabilization of Calendula officinalis L. hairy root cultures for the release of accumulated triterpenoid saponins

Triterpenoid saponins, which are glycosidic derivatives of oleanolic acid, demonstrate numerous pharmacological properties. The hairy root cultures of marigold accumulate these phytoanticipins mainly in vacuoles, which may contain up to 40% of the triterpene glycosides synthesized in the cytoplasm. Dimethyl sulfoxide, Tween 20 (T20), Tween 80 (T80), and Triton X-100 (Tx100) were used as potential surfactants, allowing for an increase in the release of saponin into the culture medium. T20 at concentrations of 0.3–1.0% (v/v) caused a higher saponin content in the liquid medium, from 52 to 61 times for CH9, 15 to 22 times higher for CC16, and seven to twelve times higher for CH2, compared to the respective control cultures. DMSO was efficient toward the CC16 line, providing a 3–7 times higher saponin content for 0.5% and 1.0% (v/v) surfactant concentrations, respectively. The suitability of Triton X-100 for triterpenoid saponin could not be determined with the method used in this experiment, and there was a serious contamination of the analyzed samples. The ultrasound method accelerated surfactant action, and only for the CH9 line did it result in an increase in the secretion of glycosides to almost three-fold in the case of T80 and over two-fold in the case of T20. The conditions that were least harmful for the roots were: ultrasound, Tween 80, and T80 interacting with US. The permeabilized cultures, after 30 days of growth in the new medium, obtained fresh biomasses similar to the control or a reduction by the maximum of one-fifth for CH9. For the second line (CC16), the growth parameters were reduced twice. Polyoxoethylene sorbitan monolaurate was found to be the most powerful surfactant, and a proposed concentration and time of action allowed for culture viability only for the CC16 line. The young parts of the root tips generated a new culture, with the growth being reduced by 77% (FW) and 82% (DW).

Topology and functional characterization of major outer membrane proteins of Treponema maltophilum and Treponema lecithinolyticum.

Numerous Treponema species are prevalent in the dysbiotic subgingival microbial community during periodontitis. The major outer sheath protein is a highly expressed virulence factor of the well-characterized species Treponema denticola. Msp forms an oligomeric membrane protein complex with adhesin and porin properties and contributes to host-microbial interaction. Treponema maltophilum and Treponema lecithinolyticum species are also prominent during periodontitis but are relatively understudied. Msp-like membrane surface proteins exist in T. maltophilum (MspA) and T. lecithinolyticum (MspTL), but limited information exists regarding their structural features or functionality. Protein profiling reveals numerous differences between these species, but minimal differences between strains of the same species. Using protein modeling tools, we predict MspA and MspTL monomeric forms to be large β-barrel structures composed of 20 all-next-neighbor antiparallel β strands which most likely adopt a homotrimer formation. Using cell fractionation, Triton X-114 phase partitioning, heat modifiability, and chemical and detergent release assays, we found evidence of amphiphilic integral membrane-associated oligomerization for both native MspA and MspTL in intact spirochetes. Proteinase K accessibility and immunofluorescence assays demonstrate surface exposure of MspA and MspTL. Functionally, purified recombinant MspA or MspTL monomer proteins can impair neutrophil chemotaxis. Expressions of MspA or MspTL with a PelB leader sequence in Escherichia coli also demonstrate surface exposure and can impair neutrophil chemotaxis in an in vivo air pouch model of inflammation. Collectively, our data demonstrate that MspA and MspTL membrane proteins can contribute to pathogenesis of these understudied oral spirochete species.

Engineered Microbial Consortium for De Novo Production of Sclareolide.

Sclareolide, a natural product with bioactive and fragrant properties, is not only utilized in the food, healthcare, and cosmetics industries but also serves as a precursor for the production of ambroxide and some bioactive compounds. Currently, there are three primary methods for producing sclareolide: direct extraction from plants, chemical synthesis using sclareol as a precursor, and the biotransformation of sclareol. Here, we established a platform for producing sclareolide through a modular coculture system with Saccharomyces cerevisiae and Cryptococcus albidus ATCC 20918. S. cerevisiae was engineered for de novo sclareol biosynthesis from glucose, while C. albidus enabled the production of sclareolide via sclareol biotransformation. To enhance the supply of sclareol, a recombinant yeast strain was constructed through metabolic engineering to produce 536.2 mg/L of sclareol. Further improvement of the coculture system for sclareolide production was achieved by incorporating Triton X-100 facilitated intermediate permeability, inoculation proportion adjustment, and culture temperature optimization. These refinements culminated in a sclareolide yield of 626.3 mg/L. This study presents a novel streamlined and efficient approach for sclareolide preparation, showcasing the potential of the microbial consortium in sustainable bioproduction.

Potentiometry in determining the antioxidant capacity of compounds of different hydrophilicity in the joint presence using mixed solvents and surfactants.

In this work, for the first time, the potentiometric method was used to determine the antioxidant capacity (AOC) of compounds of different hydrophilicity in the joint presence using mixed solvents and surfactants. The AOC of model solutions of antioxidants of different hydrophilicity was determined separately and in the joint presence in the media of phosphate buffer-surfactant and mixed solvents-surfactant, using as an example the ascorbic acid and the α-tocopherol. It was shown that the surfactant Triton X-100 is able to solubilize α-tocopherol under the selected conditions, allows to obtain reproducible and accurate results, and has less effect on the equilibrium rate of the K3[Fe(CN)6]/K4[Fe(CN)6] system. Phosphate buffer-ethanol and phosphate buffer-acetonitrile media in the 3:2 ratio with Triton X-100 (5mmol/dm3) were chosen as the analysis conditions (RSD ≤ 6%). The range of determined concentrations was (0.006-0.5)m mmol/dm3 in phosphate buffer-ethanol and (0.006-0.3)m mmol/dm3 in phosphate buffer-acetonitrile. AOC of raw materials infusions was determined in selected media. The positive correlation with the well-known Folin-Ciocalteu assay was obtained.

Exploration of micellization and phase separation nature of surfactants in metformin hydrochloride + additives media: An experimental and DFT study

The study explores the aggregation and phase separation of a newly synthesized cationic surfactant − octadecyltriethylammonium bromide (OTEAB) and triton X-100 (TX-100) respectively with/without metformin hydrochloride (MMH) drug in absence/presence of NaCl. The conductivity measurements were performed to investigate the aggregation of cationic OTEAB at temperatures between 298.15 and 323.15 K with a 5 K gap. The micelle development of OTEAB in pure state and with MMH drug has been assessed by determining the critical micelle concentration (CMC). The reduction of CMC of OTEAB has been detected owing to the introduction of the MMH drug. The CMC was further decreased in attendance of NaCl. The CMC values also demonstrate the dependency on the change of temperature in all situations investigated. The decrease of CP values of triton X-100 and MMH mixture in aqueous NaCl was attained with the augmentation of the concentration of NaCl. The negative and positive values of free energy of micellization (ΔGm0) and clouding (ΔGc0) indicate the spontaneous and nonspontaneous nature of the respective processes. The enthalpy (ΔHm0) and entropy(ΔSm0) changes of the micellization were negative (exothermic) and positive, respectively. The enthalpy (ΔHc0) and entropy(ΔSc0) changes of the clouding were negative (exothermic) and positive, respectively, at lesser and higher concentrations of NaCl. The values of ΔHm0/ΔHc0 and ΔSm0/ΔSc0 designated that both hydrophobic and electrostatic interaction are present as important forces among the components under investigation. The thermodynamics of transfer and the enthalpy-entropy compensation parameters for the two processes were also computed and explained appropriately. Herein, we have also performed the computational analysis to find out the optimized structures, HOMO-LUMO orbitals, and the band gaps of OTEAB, MMH, and OTEAB+MMH mixture.

Unveiling the role of solubilization of metformin hydrochloride assimilated in nonionic surfactants mediated mixed micellar assemblies

This study examines the distribution of an antidiabetic drug metformin hydrochloride (MNH) between water and mixed micellar environments produced by nonionic surfactants Tween 80 (TW-80) and Triton X-100 (TX-100). In our research, we described the UV–visible absorption spectra of a drug at various surfactant concentrations above and below critical micelle concentration (CMC). When employing TX-100, the absorbance spectra appeared to shift towards shorter wavelengths (blue shift) and there was an increase in absorbance intensity (hyperchromic shift), while using Tween 80, absorbance intensity increased (hyperchromic shift). We estimated the partition coefficient (Kx) from differential absorption data and utilized it to derive the free energy of partition (∆Gp). The partition coefficient in a single micellar system was found to be 7.82×106 and 1.79×106 in the presence of TW-80 and TX-100, respectively. The partition coefficient for the MNH/TW-80 system in a mixed micellar system reaches its maximum value of 1.08×107 when TX-100 is present at 1.26×106 M, indicating a considerable increase in the solubilizing power of micelles. The results showed that mixed micelles of TX-100/TW-80 were more effective for the solubilization of MNH than their individual micelles. It was crystal clear from the negative values of ∆Gb that binding is also spontaneous.

Generation of empty cell envelopes of Streptococcus pyogenes using biosurfactants

BackgroundBacterial ghost cells (BGCs) are cell envelopes that devoid of cytoplasmic and genetic contents in purpose of variable applications, including their great potential as vaccine candidates and their effectiveness as delivery systems for drugs and proteins. To our knowledge, this is the first study to produce Gram-positive BGCs by treating Streptococcus pyogenes (S. pyogenes) ATCC 19615 with Tween80 (TW80) or TritonX-100 (TX100), followed by preliminary testing of their antigenicity and safety in NIH/Ola-Hsd mice. The produced BGCs were confirmed by the presence of intact cells under a light microscope, the absence of growth signs upon re-cultivation. The transmembrane tunnels were visualized using a scanning electron microscope, and subsequently, considerable quantities of released DNA and protein were detected in the culture supernatant of the BGCs. The antigenicity of the produced BGCs was tested through three intra-nasal immunization doses followed by infection. Afterward, the opsonic activity and the IgG levels were measured, followed by a comprehensive histopathological examination for selected tissues and organs.ResultsThe sera of immunized mice exhibited a significant rise in both opsonic activity (TW80 produced BGC = 68% and TX100 produced BGC = 75%) and IgG levels (TW80 produced BGC = a threefold increase and TX100 produced BGC = a fourfold increase) when compared to the positive control group "non-immunized challenged with ATCC 19615." Histopathological analysis revealed that the BGCs produced by TW80 are relatively safer and have a less severe impact than those produced by TX100.ConclusionThe study's findings suggest that Sp-BGC/TW80 is initially effective and safe in vivo. However, further pre-clinical studies are necessary to confirm its effectiveness and ensure complete safety, specifically in terms of the absence of autoimmunity and antibody cross-reactivity with myosin proteins in human cardiac tissues.

Effect of Triton X-100 surfactant and agitation on tetramethylammonium hydroxide wet etching for microneedle fabrication

Solid silicon (Si) microneedles have many applications such as skin pretreatment to form micrometer-sized holes in the skin surface in transdermal drug delivery systems. Wet etching based on tetramethylammonium hydroxide (TMAH) is an efficient method to fabricate solid microneedles. However, it is challenging to increase the density of microneedle arrays due to the faster lateral etching than the vertical etching that requires a large initial mask size. In this work, we used wet etching based on TMAH to fabricate solid Si microneedles. One kind of nonionic surfactant, Triton X-100, was introduced into the TMAH solution to suppress the lateral etching. When Triton X-100 was added into TMAH for a given etching condition, the maximum height (attained right before the mask fell off) of microneedles could reach ∼230 μm for 600 μm square-shaped mask size and 700 μm array period, compared to microneedles of maximum 152 μm height for the same mask size and period without surfactant addition. Correspondingly, when the target heights of microneedles were the same as ∼230 μm, denser (down to 700 μm period, 600 μm mask size) microneedle arrays were achieved with the help of Triton X-100, in comparison to arrays down to 900 μm period (800 μm mask size) without surfactant addition. Furthermore, agitation by a magnetic stirring bar is important for the fabrication of dense solid Si microneedle arrays based on TMAH. The microneedle structures were rhombic pyramid in shape with Triton X-100 and agitation. But microneedle structures obtained with Triton X-100 yet without agitation were octagonal pyramid in shape with a much less steep side surface.

ZASP: A Highly Compatible and Sensitive ZnCl2 Precipitation-Assisted Sample Preparation Method for Proteomic Analysis

Universal sample preparation for proteomic analysis that enables unbiased protein manipulation, flexible reagent use, and low protein loss is required to ensure the highest sensitivity of downstream liquid chromatography-mass spectrometry (LC-MS) analysis. To address these needs, we developed a ZnCl2 precipitation-assisted sample preparation method (ZASP) that depletes harsh detergents and impurities in protein solutions prior to trypsin digestion via 10 min of ZnCl2 and methanol-induced protein precipitation at room temperature (RT). ZASP can remove trypsin digestion and LC-MS incompatible detergents such as SDS, Triton X-100, and urea at high concentrations in solution and unbiasedly recover proteins independent of the amount of protein input. We demonstrated the sensitivity and reproducibility of ZASP in an analysis of samples with 1 μg to 1000 μg of proteins. Compared to commonly used sample preparation methods such as SDC-based in-solution digestion, acetone precipitation, FASP, and SP3, ZASP has proven to be an efficient approach. Here, we present ZASP, a practical, robust, and cost-effective proteomic sample preparation method that can be applied to profile different types of samples.

Radiogenic 32P as a new dating tool to determine groundwater ages of 50–1000 years

Groundwater age has significant hydrogeological implication for groundwater renewal and solute transport. The period between 50 to 1000 years, marked by increased human activity, which is also a gap of groundwater dating. Cosmogenic 32Si with a half-life of 144±11 a, is the only viable dating tool for this range but is seldom used due to complex pretreatment and time-consuming and expensive detection. Radiogenic 32P, as a daughter isotope of 32Si, decays by emitting high-energy β- particles and is easily detected. 32Si and 32P reaches radioactive equilibrium within 3 months, meaning they maintain consistent activity levels in groundwater aged between 50 and 1000 years. Therefore, 32P can be used to date groundwater in the range of 50–1000 a. 32P is enriched and extracted using Mg(OH)2 co-precipitation method. The 32P solution is then mixed with a solvent mixture containing Triton X-100 and Permafluor and detected using an ultra-low background liquid scintillation spectrometer. The counting time ranges from 2500 to 3000 min with an efficiency of 65 %, and the 32P count peak surface is observed between channels 600 and 900. Comparing the 32P ages with the 14C and 32Si ages of groundwater samples from the Jianghan Plain, Central China, reveals that 32P ages and 14C ages exhibit similar trends, and the 32P ages and 32Si ages are approximately consistent in the range of 50–1000 a. Therefore, radiogenic 32P dating groundwater in the range of 50–1000 a is feasible, convenient and reliable.

BEAD SIZE AND RHEOLOGICAL PROPERTIES OF SEBS-BASED ELASTIC BEADS

ABSTRACT Styrene–ethylene–butylene–styrene (SEBS) is a thermoplastic elastomer that has applications in robotics and shock absorption. Although SEBS as a bulk material as well as an additive to solid composites has been extensively studied, this work focuses on developing SEBS-based beads to enhance material elasticity. SEBS bead mixtures were developed by mixing SEBS elastomer, water, and surfactant (Triton X-100) at high temperature. Stability, rheology, and microscopy of SEBS bead mixtures were studied as a function of neat SEBS concentration in SEBS elastomer, SEBS elastomer concentration, and surfactant concentration. Resulting bead mixtures were classified as creamed, homogenous and stable, or aggregated based on the mixture’s tendency to separate into layers and its ability to disperse in excess water. Microscopic studies suggest that although bead mixtures exhibit size polydispersity, the average bead size is a strong function of neat SEBS, SEBS elastomer, and surfactant concentrations. Rheological studies suggest that all the bead mixtures exhibit shear thinning behavior, and the overall viscosity of a given bead mixture is a function of both SEBS elastomer and surfactant concentration. The developed SEBS elastic beads can be used as additives to enhance the viscoelastic properties of fluid-based systems such as magnetorheological and damping fluids.

Development and Optimization of a Bromothymol Blue-Based PLA2 Assay Involving POPC-Based Self-Assemblies.

Phospholipase A2 (PLA2) is a superfamily of phospholipase enzymes that dock at the water/oil interface of phospholipid assemblies, hydrolyzing the ester bond at the sn-2 position. The enzymatic activity of these enzymes differs based on the nature of the substrate, its supramolecular assemblies (micelle, liposomes), and their composition, reflecting the interfacial nature of the PLA2s and requiring assays able to directly quantify this interaction of the enzyme(s) with these supramolecular assemblies. We developed and optimized a simple, universal assay method employing the pH-sensitive indicator dye bromothymol blue (BTB), in which different POPC (3-palmitoyl-2-oleoyl-sn-glycero-1-phosphocholine) self-assemblies (liposomes or mixed micelles with Triton X-100 at different molar ratios) were used to assess the enzymatic activity. We used this assay to perform a comparative analysis of PLA2 kinetics on these supramolecular assemblies and to determine the kinetic parameters of PLA2 isozymes IB and IIA for each supramolecular POPC assembly. This assay is suitable for assessing the inhibition of PLA2s with great accuracy using UV-VIS spectrophotometry, being thus amenable for screening of PLA2 enzymes and their substrates and inhibitors in conditions very similar to physiologic ones.

Fibrotic liver extracellular matrix induces cancerous phenotype in biomimetic micro-tissues of hepatocellular carcinoma model

BackgroundDespite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma (HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of prolonged liver fibrosis, resulting in the formation of an intricate premalignant microenvironment. The accumulation of extracellular matrix (ECM) is a hallmark of premalignant microenvironment. Given the critical role of different matrix components in regulating cell phenotype and function, this study aimed to elucidate the interplay between the fibrotic matrix and malignant features in HCC. MethodsLiver tissues from both control (normal) and carbon tetrachloride (CCl4)-induced fibrotic rats were decellularized using sodium dodecyl sulfate (SDS) and Triton X-100. The resulting hydrogel from decellularized ECM was processed into micro-particles via the water-in-oil emulsion method. Micro-particles were subsequently incorporated into three-dimensional liver biomimetic micro-tissues (MTs) comprising Huh-7 cells, human umbilical vein endothelial cells (HUVECs), and LX-2 cells. The MTs were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay at day 11, immunofluorescence staining, immunoblotting, and spheroid migration assay at day 14 after co-culture. ResultsFibrotic matrix from CCl4-treated rat livers significantly enhanced the growth rate of the MTs and their expression of CCND1 as compared to the normal one. Fibrotic matrix, also induced the expression of epithelial-to-mesenchymal transition (EMT)-associated genes such as TWIST1, ACTA2, MMP9, CDH2, and VIMENTIN in the MTs as compared to the normal matrix. Conversely, the expression of CDH1 and hepatic maturation genes HNF4A, ALB, CYP3A4 was decreased in the MTs when the fibrotic matrix was used. Furthermore, the fibrotic matrix increased the migration of the MTs and their secretion of alpha-fetoprotein. ConclusionsOur findings suggest a regulatory role for the fibrotic matrix in promoting cancerous phenotype, which could potentially accelerate the progression of malignancy in the liver.

A simple optical sensor for the simultaneous determination of trace amounts of silver and gold nanoparticles in water by cloud point microextraction

Nowadays, metal nanoparticles are widely used in many fields due to their excellent properties. This has led to an increasing presence of these species in water, which, although in very low concentrations, can cause damage to ecosystems. These nanomaterials are currently considered as emerging pollutants in water, so there is a need to know their concentrations. In particular, gold and silver nanoparticles are among the most widely used. Therefore, the development of simple methods that allow their determination even at trace concentrations is of interest. In this work, a method has been developed for the simultaneous determination of low concentrations of silver and gold nanoparticles (AgNPs and AuNPs, respectively) in water by cloud point microextraction using Triton X-114 as extractant and UV–Vis molecular spectrophotometry. It is a simple, environmentally friendly method with an easy-to-use portable instrument that requires only a microdrop of the extracted phase for the analysis of both species. Ther detection limits reached were 17 and 15 µg L−1 for AgNPs and AuNPs, respectively. The enrichment factors were 160 for AgNPs and 165 for AuNPs.

Design of green DES-based aqueous two phase systems for lipolytic enzymes extraction

This study pioneers a novel green and cost-effective separation process using reline deep eutectic solvent (DES) for extracting Thermomyces lanuginosus lipase (TlL) from aqueous solutions. For the first time, it has been shown that various potassium organic salts combined with reline DES can induce phase separation in aqueous solutions of the biodegradable non-ionic surfactant Tergitol 15-S9, presenting a groundbreaking alternative to the banned surfactant Triton X-100. This work uniquely applies three different equations to describe the experimental solubility curves. The selection of reline DES is particularly innovative as it avoids significant deactivation effects with three commercial lipases: Candida antarctica lipases A and B, and TlL, with the latter undergoing a detailed study. This includes an unprecedented analysis of its thermodeactivation kinetics in reline DES aqueous solutions. Additionally, a novel investigation of immiscibility regions at different temperatures, involving comprehensive Tie-Line characterizations (slope and length), preceded the evaluation of lipase extraction. Remarkably, the study demonstrates that optimizing the feed composition to contain higher water concentrations can achieve exceptionally high extraction efficiencies (approximately 90%), suggesting a transformative potential for future applications in real aqueous cultures.

Quantification of tungsten in real samples spectrophotometrically using optical sensor

A new methodology was devised to detect tungsten in aqueous samples, employing a sensor membrane crafted by physically incorporating a tungsten-selective compound, 5-(2 -bromophenylazo)-6-hydroxypyrimidine-2,4-dione (BPAHPD), into a plasticized poly(vinyl chloride). The addition of Triton X-114 was observed to be advantageous in augmenting the absorption of HWO4− ions from the liquid phase into the membrane phase, thereby amplifying sensor absorption intensity. The method established a linear dynamic span of 15–350 ng mL−1 for W(VI), exhibiting a notable correlation coefficient of 0.9992. Quantification and detection limits were determined to be 14.85 and 4.45 ng mL−1, correspondingly, under optimized conditions. The Sandell responsiveness and molar absorptivity were calculated at 0.0455 ng cm−2 and 4.04 × 108 L/mol cm−1, respectively. The sensor demonstrated superb selectivity, as demonstrated by the negligible influence of potential interfering species. Statistical analyses, encompassing variance ratio F-tests and Student’s t-tests, indicated no substantial differences. The efficacy of the methodology was authenticated through the scrutiny of genuine samples, with statistical comparison to the ICP-AES method.

Novel Triton X-114 coated Fe3O4 magnetic nanoparticle adsorbent for Cd(II) and Co(II) ions preconcentration in honey samples using magnetic solid-phase extraction: Determination by ICP-OES

This work focuses on creating and studying the effectiveness of Fe3O4 nanoparticles coated with (3-aminopropyl)triethoxysilane (APTES) and polyethylene glycol tert-octylphenyl ether (TX-114), respectively, as adsorbent. Fe3O4@APTES@TX-114 adsorbent extracts heavy metal cations Cd2+ and Co2+ using a magnetic solid phase extraction technique. The synthesized magnetic nanoparticles (MNPs) were characterized by Field Emission Scanning Electron Microscope (FESEM), Energy Dispersive X-Ray Analysis (EDX), Fourier Transform Infrared Spectroscopy (FT-IR), and Thermogravimetric Analysis (TGA). After this stage, Cd2+ and Co2+ determinations were made with the ICP-OES device. The optimum conditions for recovering Cd2+ and Co2+ cations were pH 7, an adsorbent dosage of 2 g/L, and a contact time of 120 min. While the method was compatible with Langmuir and Freundlich isotherm models, the pseudo-second-order kinetic equation was more compatible than the pseudo-first-order. Acidic and basic media were investigated for the desorption of Cd2+ and Co2+ ions from the adsorbent. At the same time, maximum recovery was achieved for both cations at a concentration of 1.5 mol/L HCl, but no significant recovery was obtained in the NaOH medium. The common ion study observed that Cd2+ and Co2+ cations were recovered over 95 % in the presence of many cations and anions at different concentrations. The relative standard deviation (RSD), limit of quantification (LOQ), limit of detection (LOD), and enrichment factor (EF) were calculated as 1.40 %, 2.963 µg/L, 0.889 µg/L, 20.99 for Cd2+ and 3.05 %, 1.88 µg/L, 0.564 µg/L, 109.41 for Co2+, respectively. The magnetic solid phase extraction method realized quantitative recovery of Cd2+ and Co2+ cations in different honey samples. It was found that the heavy metal contents in honey samples from the Sakarya region are below permissible levels.