Membrane Extraction Research Articles

Development of Polarity-Reversed Endometrial Epithelial Organoids.

The uterine epithelium is composed of a single layer of hormone responsive polarized epithelial cells that line the lumen and form tubular glands. Endometrial epithelial organoids (EEO) can be generated from uterine epithelia and recapitulate cell composition and hormone responses in vitro. As such, the development of EEO represents a major advance for facilitating mechanistic studies in vitro. However, a major limitation for the use of EEO cultured in basement membrane extract and other hydrogels is the inner location of apical membrane, thereby hindering direct access to the apical surface of the epithelium to study interactions with the embryo or infectious agents such as viruses and bacteria. Here, a straightforward strategy was developed that successfully reverses the polarity of EEO. The result is an apical-out organoid that preserves a distinct apical-basolateral orientation and remains responsive to ovarian steroid hormones. Our investigations highlight the utility of polarity-reversed EEO to study interactions with E. coli and blastocysts. This method of generating apical-out EEO lays the foundation for developing new in vitro functional assays, particularly regarding epithelial interactions with embryos during pregnancy or other luminal constituents in a pathological or diseased state.

Targeted chemotherapy via HER2-based chimeric antigen receptor (CAR) engineered T-cell membrane coated polymeric nanoparticles

Cell membrane-derived nanoparticles (NPs) have recently gained popularity due to their desirable features in drug delivery such as mimicking properties of native cells, impeding systemic clearance, and altering foreign body responses. Besides NP technology, adoptive immunotherapy has emerged due to its promise in cancer specificity and therapeutic efficacy. In this research, we developed a biomimetic drug carrier based on chimeric antigen receptor (CAR) transduced T-cell membranes. For that purpose, anti-HER2 CAR-T cells were engineered via lentiviral transduction of anti-HER2 CAR coding lentiviral plasmids. Anti-HER2 CAR-T cells were characterized by their specific activities against the HER2 antigen and used for cell membrane extraction. Anti-cancer drug Cisplatin-loaded poly (D, l-lactide-co-glycolic acid) (PLGA) NPs were coated with anti-human epidermal growth factor receptor 2 (HER2)-specific CAR engineered T-cell membranes. Anti-HER2 CAR-T-cell membrane-coated PLGA NPs (CAR-T-MNPs) were characterized and confirmed via fluorescent microscopy and flow cytometry. Membrane-coated NPs showed a sustained drug release over the course of 21 days in physiological conditions. Cisplatin-loaded CAR-T-MNPs also inhibited the growth of multiple HER2+ cancer cells in vitro. In addition, in vitro uptake studies revealed that CAR-T-MNPs showed an increased uptake by A549 cells. These results were also confirmed via in vivo biodistribution and therapeutic studies using a subcutaneous lung cancer model in nude mice. CAR-T-MNPs localized preferentially at tumor areas compared to those of other studied groups and consisted of a significant reduction in tumor growth in tumor-bearing mice. In Conclusion, the new CAR modified cell membrane-coated NP drug-delivery platform has demonstrated its efficacy both in vitro and in vivo. Therefore, CAR engineered membrane-coated NP system could be a promising cell-mimicking drug carrier that could improve therapeutic outcomes of lung cancer treatments.

The effect of extracellular matrix on the precision medicine utility of pancreatic cancer patient-derived organoids.

The use of patient-derived organoids (PDOs) to characterize therapeutic sensitivity and resistance is a promising precision medicine approach, and its potential to inform clinical decisions is now being tested in several large multiinstitutional clinical trials. PDOs are cultivated in the extracellular matrix from basement membrane extracts (BMEs) that are most commonly acquired commercially. Each clinical site utilizes distinct BME lots and may be restricted due to the availability of commercial BME sources. However, the effect of different sources of BMEs on organoid drug response is unknown. Here, we tested the effect of BME source on proliferation, drug response, and gene expression in mouse and human pancreatic ductal adenocarcinoma (PDA) organoids. Both human and mouse organoids displayed increased proliferation in Matrigel compared with Cultrex and UltiMatrix. However, we observed no substantial effect on drug response when organoids were cultured in Matrigel, Cultrex, or UltiMatrix. We also did not observe major shifts in gene expression across the different BME sources, and PDOs maintained their classical or basal-like designation. Overall, we found that the BME source (Matrigel, Cultrex, UltiMatrix) does not shift PDO dose-response curves or drug testing results, indicating that PDO pharmacotyping is a robust approach for precision medicine.

High-Temporal-Resolution In Situ Sensor for Oceanic CO2 Isotope Measurement Enabling Multidimensional Isotope Tracing Analysis (R13C, R18O, and R17O) via Laser Absorption Spectroscopy.

Combined in situ analysis of oceanic CO2 concentrations and diverse C and O isotope characteristics can offer a unique perspective with multiple isotopic tracing dimensions for identifying marine biogeochemical processes. Applying this strategy in marine environments is urgently required, yet it faces inherent challenges in terms of existing analytical methods and instruments, e.g., a lack of in situ sensors, limited detectable isotope variety, and low-temporal-resolution data. Here, we report an underwater in situ dissolved CO2 isotope sensor based on mid-infrared tunable diode laser absorption spectroscopy (MIR-TDLAS) and membrane extraction technology. Through the proposed targeted strategies, the sensor is capable of providing high-temporal-resolution in situ measurement of all monosubstituted isotopes of dissolved CO2 (16O13C16O, 18O12C16O, and 17O12C16O) at marine background concentrations. The sensor is demonstrated to provide comparable precision to that of isotope ratio mass spectrometry. At 400 ppmv, the precision for R13C, R18O, and R17O could achieve 0.084, 0.042, and 0.013‰, respectively, for a 1 s integration time. By enabling a high-frequency in situ analysis in fixed-point time-series field deployment, a 17O anomaly with strong regularity is observed, which is not obvious in 18O and 13C, and therefore, the superiority of the proposed multidimensional in situ isotope tracing strategy is demonstrated. The developed sensor has great potential to open up new prospects for advancing marine carbon research.

Multiplexed Live-Cell Imaging for Drug Responses in Patient-Derived Organoid Models of Cancer.

Patient-derived organoid (PDO) models of cancer are a multifunctional research system that better recapitulates human disease as compared to cancer cell lines. PDO models can be generated by culturing patient tumor cells in extracellular basement membrane extracts (BME) and plating them as three-dimensional domes. However, commercially available reagents that have been optimized for phenotypic assays in monolayer cultures often are not compatible with BME. Herein, we describe a method to plate PDO models and assess drug effects using an automated live-cell imaging system. In addition, we apply fluorescent dyes that are compatible with kinetic measurements to quantify cell health and apoptosis simultaneously. Image capture can be customized to occur at regular time intervals over several days. Users can analyze drug effects in individual Z-plane images or a Z Projection of serial images from multiple focal planes. Using masking, specific parameters of interest are calculated, such as PDO number, area, and fluorescence intensity. We provide proof-of-concept data demonstrating the effect of cytotoxic agents on cell health, apoptosis, and viability. This automated kinetic imaging platform can be expanded to other phenotypic readouts to understand diverse therapeutic effects in PDO models of cancer.

Loss of the yeast transporter Agp2 upregulates the pleiotropic drug-resistant pump Pdr5 and confers resistance to the protein synthesis inhibitor cycloheximide.

The transmembrane protein Agp2, initially shown as a transporter of L-carnitine, mediates the high-affinity transport of polyamines and the anticancer drug bleomycin-A5. Cells lacking Agp2 are hyper-resistant to polyamine and bleomycin-A5. In these earlier studies, we showed that the protein synthesis inhibitor cycloheximide blocked the uptake of bleomycin-A5 into the cells suggesting that the drug uptake system may require de novo synthesis. However, our recent findings demonstrated that cycloheximide, instead, induced rapid degradation of Agp2, and in the absence of Agp2 cells are resistant to cycloheximide. These observations raised the possibility that the degradation of Agp2 may allow the cell to alter its drug resistance network to combat the toxic effects of cycloheximide. In this study, we show that membrane extracts from agp2Δ mutants accentuated several proteins that were differentially expressed in comparison to the parent. Mass spectrometry analysis of the membrane extracts uncovered the pleiotropic drug efflux pump, Pdr5, involved in the efflux of cycloheximide, as a key protein upregulated in the agp2Δ mutant. Moreover, a global gene expression analysis revealed that 322 genes were differentially affected in the agp2Δ mutant versus the parent, including the prominent PDR5 gene and genes required for mitochondrial function. We further show that Agp2 is associated with the upstream region of the PDR5 gene, leading to the hypothesis that cycloheximide resistance displayed by the agp2Δ mutant is due to the derepression of the PDR5 gene.

Cytotoxicity of dilutions of bioceramic materials in stem cells of human exfoliated deciduous teeth.

Several materials have been developed to preserve pulp vitality. They should have ideal cytocompatibility characteristics to promote the activity of stem cells of human exfoliated deciduous teeth (SHED) and thus heal pulp tissue. To evaluate the cytotoxicity of different dilutions of bioceramic material extracts in SHED. SHED were immersed in αMEM + the material extract according to the following experimental groups: Group 1 (G1) -BBio membrane, Group 2 (G2) - Bio-C Repair, Group 3 (G3) - MTA Repair HP, Group 4 (G4) - TheraCal LC, and Group 5 (G5) - Biodentine. Positive and negative control groups were maintained respectively in αMEM + 10% FBS and Milli-Q Water. The methods to analyze cell viability and proliferation involved MTT and Alamar Blue assays at 24, 48, and 72H after the contact of the SHED with bioceramic extracts at 1:1 and 1:2 dilutions. Data were analyzed by the three-way ANOVA, followed by Tukey's test (p<0.05). At 1:1 dilution, SHED in contact with the MTA HP Repair extract showed statistically higher cell viability than the other experimental groups and the negative control (p<0.05), except for TheraCal LC (p> 0.05). At 1:2 dilution, BBio Membrane and Bio-C showed statistically higher values in intra- and intergroup comparisons (p<0.05). BBio Membrane, Bio-C Repair, and Biodentine extracts at 1:1 dilution showed greater cytotoxicity than 1:2 dilution in all periods (p<0.05). MTA HP Repair showed the lowest cytotoxicity even at a 1:1 dilution. At a 1:2 dilution, the SHED in contact with the BBio membrane extract showed high cell viability. Thus, the BBio membrane would be a new non-cytotoxic biomaterial for SHED. Results offer possibilities of biomaterials that can be indicated for use in clinical regenerative procedures of the dentin-pulp complex.

Extraction of Alpha-cypermethrin Pesticides from Aqueous Solution by Emulsion Liquid Membrane

This research examines the emulsion liquid membrane (ELM) extraction efficiency and stability to abstraction of alpha-cypermethrin pesticides from aqueous solution when blended surfactants are used in the liquid membrane composition. The stability of the membrane was examined in the aspect of the emulsion breakdown time and distribution of droplet size. The ingredients employed in ELM were Span80 (sorbitan monooleate) and Tween80 (polyoxyethylene sorbitan monooleate) as surfactants and a mixture of kerosene, sunflower oil (3:7) as diluents and stripping agent was used sodium hydroxide (NaOH). Some parameters were assessed, including the value of Hydrophilic Lipophilic Balance (HLB), blended surfactant concentration, homogenizer speed, emulsification time, and NaOH concentration as a stripping agent. According to the results, a stable emulsion was created at HLB (5.37), blended surfactant 3 % (w/v), 12,700 rpm of homogenizer speed, 9 min of emulsification time, and 0.05 N of NaOH. Under these circumstances, 89.2 % of the alpha-cypermethrin was extracted with 1.8 % emulsion breakage. As a result, the combinations of (Span80 with Tween80) have enormous potential as surfactants and they can improve the stability of the emulsion. Additionally, completed was mass transfer investigation and extraction kinetics

Generation and characterization of nanobodies targeting GPCR.

G protein-coupled receptors (GPCRs) are a large family of cell membrane proteins that are important targets for drug discovery. Nanobodies, also known as VHH (variable domains of heavy chain-only antibodies, HcAbs) antibodies, are small antibody fragments derived from camelids that have gained significant attention as potential therapeutics for targeting GPCRs due to their advantages over conventional antibodies. However, there are challenges in developing nanobodies targeting GPCRs, among which epitope accessibility is the most significant because the cell membrane partially shields the GPCR surface. We developed a universal protocol for making nanobodies targeting GPCRs using the cell membrane extract of GPCR-overexpressing HEK293 cells as the llama/alpaca immunization antigen. We constructed an immune VHH library and identified nanobodies by phage display bio-panning. The monoclonal nanobodies were recombinantly expressed in Escherichia coli (E. coli) and purified to characterize their binding potency.

Comparative analyses of adsorbed circulating proteins in the PMMA and PES hemodiafilters in patients on predilution online hemodiafiltration.

Acute and chronic inflammation are common in patients with end-stage kidney disease (ESKD). So, the adsorption of pro-inflammatory cytokines by the hollow fiber of the dialysis membrane has been expected to modify the inflammatory dysregulation in ESKD patients. However, it remains to be determined in detail what molecules of fiber materials can preferably adsorb proteins from the circulating circuit. We aimed this study to analyze directly the adsorbed proteins in the polymethyl methacrylate (PMMA) and polyethersulfone (PES) membranes in patients on predilution online hemodiafiltration (OL-HDF). To compare the adsorbed proteins in the PMMA and PES hemodiafilters membrane, we initially performed predilution OL-HDF using the PES (MFX-25Seco) membrane while then switched to the PMMA (PMF™-A) membrane under the same condition in three patients. We extracted proteins from the collected hemodiafilters by extraction, then SDS-PAGE of the extracted sample, protein isolation, in-gel tryptic digestion, and nano-LC MS/MS analyses. The concentrations of adsorbed proteins from the PMMA and PES membrane extracts were 35.6±7.9 μg/μL and 26.1±9.2 μg/μL. SDS-PAGE analysis revealed distinct variations of adsorbed proteins mainly in the molecular weight between 10 to 25 kDa. By tryptic gel digestion and mass spectrometric analysis, the PMMA membrane exhibited higher adsorptions of β2 microglobulin, dermcidin, retinol-binding protein-4, and lambda-1 light chain than those from the PES membrane. In contrast, amyloid A-1 protein was adsorbed more potently in the PES membrane. Western blot analyses revealed that the PMMA membrane adsorbed interleukin-6 (IL-6) approximately 5 to 118 times compared to the PES membrane. These findings suggest that PMMA-based OL-HDF therapy may be useful in controlling inflammatory status in ESKD patients.

Biological lipid hydration: distinct mechanisms of interfacial water alignment and charge screening for model lipid membranes.

Studying lipid monolayers as model biological membranes, we demonstrate that water molecules interfacing with different model membranes can display preferential orientation for two distinct reasons: due to charges on the membrane, and due to large dipole fields resulting from zwitterionic headgroups. This preferential water orientation caused by the charge or the dipolar field can be effectively neutralized to net-zero water orientation by introducing monolayer counter-charges (i.e. lipids with oppositely charged headgroups). Following the Gouy-Chapman model, the effect of monolayer surface charge on water orientation is furthermore strongly dependent on the electrolyte concentration and thus on the counterions in solution. In contrast, the effect of ions in the subphase on the dipolar alignment of water is zero. As a result, the capability of monolayer counter-charges to null the effect of dipolar orientation is strongly electrolyte-dependent. Notably, the different effects are additive for mixed charged/zwitterionic lipid systems occurring in nature. Specifically, for an E. coli lipid membrane extract consisting of both zwitterionic and negatively charged lipids, the water orientation can be explained by the sum of the constituents. Our results can be quantitatively reproduced using Gouy-Chapman theory, revealing the relatively straightforward electrostatic effects on the hydration of complex membrane interfaces.

Extraction of acid orange 7 from water by emulsion liquid membrane using Tri-dodecyl-amine as extractant

The main objective of this study was to investigate the feasibility of extracting Acid Orange 7 (AO7), a complex reactive dye, from aqueous solutions using a stabilized emulsion liquid membrane (ELM) technique. The ELM formulation comprised tri-dodecylamine (TDA) as the extracting agent, kerosene as the diluting medium, and SPAN 80 as the surfactant. We systematically examined various experimental variables that affect the extraction of AO7. These variables included the concentrations of the extractant and surfactant, internal phase concentrations, the ratio of the membrane phase to the internal phase, the choice of diluting medium and its grade, stirring speed, treatment ratio, and the initial concentration of AO7. Furthermore, we investigated the impact of sodium chloride concentration in the external phase. Our research findings indicate that the optimal conditions for AO7 extraction using the ELM method were as follows: a TDA concentration of 3% (w/w), a SPAN 80 concentration of 9% (w/w), an oil-to-aqueous phase ratio (O/A) of 1:1, kerosene as the diluting medium, H2SO4 as the internal phase, a stirring speed of 200 rpm, a treatment ratio of 1:10, and an initial AO7 concentration of 10 mg/L. Under these precise conditions, an exceptional extraction efficiency of 98.49% was achieved in just 15 min. Under these carefully controlled conditions, an exceptional extraction efficiency of 98.49% was achieved in just 15 min. Consequently, the emulsified liquid membrane extraction process demonstrates significant potential as an advanced separation technique for removing hazardous dyes, particularly in dilute solutions.

Extraction/transportation of Co2+ metal ions with bis(2-ethylhexyl) phosphate dissolved in kerosene using a multidropped liquid membrane technique.

The transport properties of Co2+ ions from the aqueous donor phase to aqueous acceptor phase with the recently developed multidroplet liquid membrane (MDLM) extraction system were studied. This system serves as a continuous process for the transportation of ions and requires fewer reagents for starting and conducting the procedure. Moreover, the procedure results in fewer waste chemicals and mixtures in comparison to traditional extraction methods. During extraction, bis(2-ethylhexyl) phosphate (D2EHPA) was used as a carrier material and 5% potassium thiocyanate (KSCN) solution was used to obtain a colored complex for UV-Vis detection. By means of several experiments, the optimum D2EHPA concentration, pH range for both donor and acceptor phases, and temperature range effect on transport kinetics were investigated. In the extraction of cobalt ions with the MDLM system, the activation energy was calculated as Ea = 13.80 kcal mol-1, and it was found that the extraction was chemically controlled since it was greater than 10 kcal mol-1.

Subcellular RNA distribution and its change during human embryonic stem cell differentiation

The spatial localization of RNA within cells is closely related to its function and also involved in cell fate determination. However, the atlas of RNA distribution within cells and dynamic changes during the developmental process are largely unknown. In this study, five subcellular components, including cytoplasmic extract, membrane extract, soluble nuclear extract, chromatin-bound nuclear extract, and cytoskeletal extract, were isolated and the rules of subcellular RNA distribution in human embryonic stem cells (hESCs) and its change during hESC differentiation are summarized for the first time. The overall distribution patterns of coding and non-coding RNAs are revealed. Interestingly, some developmental genes are found to be transcribed but confined to the chromatin in undifferentiated hESC. Unexpectedly, alternative splicing and polyadenylation endow spatial heterogeneity among different isoforms of the same gene. Finally, the dynamic pattern of RNA distribution during hESC differentiation is characterized, which provides new clues for a comprehensive understanding hESC pluripotency and differentiation.

A stochastic variance reduction gradient-based GSO-ANFIS optimized method for maximum power extraction of proton exchange membrane fuel cell

Proton Exchange Membrane Fuel Cells (PEMFCs) play a pivotal role in the clean energy landscape, yet their efficiency is contingent upon effective power optimization. This paper presents Maximum PowerPoint Tracking (MPPT) control schemes for PEMFCs, focusing on a ground-breaking methodology. Traditional MPPT controllers are instrumental in maintaining optimal performance; however, they often struggle with dynamic operating conditions. In response to this challenge, this research work presents a pioneering MPPT control scheme employing a stochastic variance reduction gradient system. The novelty of this approach lies in its fusion with the Glow Swarm Optimization (GSO) and the Adaptive Neuro Fuzzy Inference System (ANFIS), resulting in a robust hybrid controller. In the pursuit of optimizing the PEMFC system, the proposed GSO-ANFIS controller is subjected to rigorous testing under dynamic variations in both PEMFC temperature and load. Notably, PEMFCs, due to fluctuations in pressure and temperature, exhibit stochastic behaviour, forming a Gaussian surface. In this research, the popular Perturb and Observe (P&O) and Incremental Conductance methods are evaluated alongside the newly introduced GSO-ANFIS model. The proposed GSO-ANFIS controller outperforms its counterparts, showcasing an impressive accuracy level of 89.97%. In contrast, the Artificial Neural Network (ANN) achieves 80.33% accuracy, and the standalone ANFIS controller attains 86.5% accuracy. This disparity underscores the efficacy and potential of the novel hybrid approach, which not only adeptly handles the stochastic nature of PEMFCs but also significantly enhances accuracy in power optimization. This research not only contributes a valuable addition to the field of MPPT control but also offers a promising trajectory for the future development of efficient and reliable PEMFC systems.

Parallel artificial liquid membrane extraction coupled with UPLC-ESI-MS/MS method for high-throughput quantitation of repaglinide in diabetic patients

A high-throughput therapeutic monitoring method was developed for repaglinide (RPG) in diabetic patients, combining parallel artificial liquid membrane extraction (PALME) with ultraperformance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). PALME was performed using a 96-well donor plate comprising a donor solution containing a plasma sample, 50 mM phosphate buffer (pH = 8.0), and cetirizine (CTZ) as internal standard. A polypropylene (PP) porous membrane served as a selective support for the liquid membrane (SLM), preventing nonspecific binding produced by other membranes. The extraction was accomplished across SLM made of PP membrane with dodecyl acetate and 1 % trioctylamine (w/w), and the acceptor solution comprised DMSO and 200 mM formic acid (50:50, v/v). The simple workflow for PALME provided analyte enrichment, highly efficient sample cleanup, high throughput analysis, and excellent reproducibility. Method validation met FDA criteria, with a linear plasma calibration range (0.1–100 ng mL−1, r = 0.9995) and a lower limit of quantitation (LLOQ) of 0.1 ng mL−1. Recovery results at 98.9 % affirmed method reliability. The ability to analyze 198 samples per hour, coupled with a reduced amount of solvents, underscores the method's high throughput and eco-friendly profile. The PALME-UPLC-ESI-MS/MS method was successfully applied to therapeutic drug monitoring of RPG in diabetic patients following 2 mg RPG tablet administration, establishing its effectiveness.

Formulation and Evaluation of Emulgel loaded with leaf extract of Artemisia princeps for its Anti-Inflammatory potential

Indian Systems of Medicine have their roots in ayurveda. The potential of naturally occurring bioactive components is of tremendous interest to researchers. Different plant extracts have remedial properties.North India, Japan, China, Korea and Nepal are among the countries where Artemisia princeps can be found.Apigenin, diosmetin, eriiodicytol, eupafolin, kaempferol, vitex, jaceosidine, and quercetins have all been discovered in plant extracts of Artemisia princeps. It is used to treat fever, apoptosis, pain, ulcers, inflammation, and other conditions as well as Diarrhoea and Asthma.Recent in-vivo and in-vitro research have shown that it has anti-inflammatory and analgesic properties. The objectives of proposed research work are to formulate, evaluate and optimize the Emulgel loaded with optimised extract of Artemisia princepsand assess its In-Vitro Anti-Inflammatory Activity. Collected leaves of Artemisia princeps was extracted using hydro-alcoholic solvent. On the basis of their Phytochemical evaluation, the extract was optimized. Additional 6 batches of Emulgel were created utilizing a chosen extract. The prepared Emulgel was assessed for pH, spreadability, viscosity, consistency, appearance, colour, infrared spectral assignment, and appearance. Using the HRBC Membrane stabilisation assay, extract and formulation were tested for in vitro anti-inflammatory efficacy. The suggested study project's findings support the potential for future topical application of the newly developed Emulgel, which contains Artemisia princeps leaf extract, to reduce inflammation.

Flower-like calix[6]arene-based covalent organic framework for membrane extraction of sulfonamides in animal-derived food through host-guest interaction prior to determination with ultra-high performance liquid chromatography-tandem mass spectrometry

Supramolecular macrocycle-based covalent organic frameworks (COFs) are promising adsorbents for adsorption of hazards due to their host-guest recognition property. However, most supramolecular macrocycles are conformationally flexible, making them challenging to introduce into COFs. In this work, a calix[6]arene-based COF (CX6-BD COF) was fabricated with a unique flower-like morphology and high crystallinity. Especially, the cavity of CX6 exhibited host-guest inclusion interaction for sulfonamides (SAs), which was verified by quantum chemistry calculation. The integration of the porosity of COFs with the recognition cavity of CX6 made CX6-BD COF display excellent enrichment performance for SAs, with good enrichment factors (EFs) between 77 and 96. The material was employed as an adsorbent for COF membrane filter extraction, coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to simultaneously enrich and determine seven SAs in animal-derived food. The analytical method showed a wide linear range (0.01–100 µg/L and 0.05–100 µg/L) and low detection limits (3–10 ng/L). The established method was successfully applied to sensitively determine SAs in chicken, pork and beef samples, which achieved satisfactory recoveries (73.8–113%). These results demonstrated CX6-BD COF has good application potential in determination of trace and ultra-trace SAs in complex food matrices as an adsorbent.

Membrane extraction in native lipid nanodiscs reveals dynamic regulation of Cdc42 complexes during cell polarization

Embryonic development requires the establishment of cell polarity to enable cell fate segregation and tissue morphogenesis. This process is regulated by Par complex proteins, which partition into polarized membrane domains and direct downstream polarized cell behaviors. The kinase aPKC (along with its cofactor Par6) is a key member of this network and can be recruited to the plasma membrane by either the small GTPase Cdc42 or the scaffolding protein Par3. Although in vitro interactions among these proteins are well established, much is still unknown about the complexes they form during development. Here, to enable the study of membrane-associated complexes ex vivo, we used a maleic acid copolymer to rapidly isolate membrane proteins from single C. elegans zygotes into lipid nanodiscs. We show that native lipid nanodisc formation enables detection of endogenous complexes involving Cdc42, which are undetectable when cells are lysed in detergent. We found that Cdc42 interacts more strongly with aPKC/Par6 during polarity maintenance than polarity establishment, two developmental stages that are separated by only a few minutes. We further show that Cdc42 and Par3 do not bind aPKC/Par6 simultaneously, confirming recent in vitro findings in an ex vivo context. Our findings establish a new tool for studying membrane-associated signaling complexes and reveal an unexpected mode of polarity regulation via Cdc42.

A new approach for cobalt (II) removal from simulated wastewater using electro membrane extraction with a flat sheet supported liquid membrane

The aim of this work was to efficiently remove cobalt (Co) from aqueous solutions by using a novel Electromembrane Extraction (EME) technique. This novel electrochemical cell design featured two distinct glass chambers, incorporating a Supported Liquid Membrane (SLM) composed of a polypropylene flat membrane saturated with 1-octanol and a carrier substance, as well as electrodes constructed from graphite and stainless steel. The investigation covered an exploration of various effective parameters like, carrier type, voltage across the cell, donor solution pH, and the initial Co concentration, with the overarching goal of comprehending their individual effect on Co removal efficiency. Notably, two different carriers, tris(2-ethylhexyl) phosphate (TEHP) and bis(2-ethylhexyl) phosphate (DEHP), were systematically evaluated in combination with 1-octanol. The findings underscored the pivotal role of the cell voltage in significantly enhancing the mass transfer rate of cobalt across the membrane, thereby advancing the effectiveness of the removal process. After a comprehensive optimization process, the optimal operating conditions were established as follows: employing 1-octanol with 1.0 % v/v bis(2-ethylhexyl) phosphate as a carrier, applying a voltage of 60 V, maintaining an initial pH of 5, utilizing an initial cobalt concentration of 15 mg/L, conducting an extraction for 6 h, and employing a stirring rate of 1000 rpm. Remarkably, these conditions led to the attainment of an impressive removal efficiency of 87 %. In stark contrast, when no voltage was applied, the removal efficiency did not surpass 40 %. This underscores the pivotal role of the applied voltage in enhancing the cobalt removal process under the specified conditions.