Membrane Orientation Research Articles

Engineered surface design of recognition site-ordered biomimetic sensor for efficient detection of circulating tumor cells

The oriented assembly of cell membrane coating plays an important role in advancing the application of this strategy in biomedical fields, particularly in detecting circulating tumor cells (CTCs). Unfortunately, there is a formidable challenge in achieving effective membrane orientation during the coating process owing to the asymmetric properties of cell membranes. Herein, magnetic vesicles released by tumor cells were designed to break down these barriers in the same way that microvesicles are actively secreted from cells, which completely inherited the orientation and characteristics of the parent cell membranes, exhibiting a satisfactory self-targeting ability for homologous cells. To cope with the complex application environment, spatially ordered aptamers were integrated into magnetic vesicles and combined with catalytic hairpin assembly (CHA) technology to construct a recognition site-ordered biomimetic sensor for high-performance detection of CTCs. In this strategy, the ordered arrangement of membrane proteins and aptamers markedly improved capture efficiency of traditional biomimetic strategy for CTCs. Additionally, CHA-induced fluorescence and colorimetric analysis ensured the detection accuracy and sensitivity, with a linear range of 0 to 104 cells mL−1 and a low detection limit of 3 cells mL−1 for fluorometry and 6 cells mL−1 for colorimetry. Overall, the biomimetic sensor offered broader possibilities for detecting rare CTCs and provided new insight to expand the application of cell membrane biomimetic strategies in biomedicine.

Dramatic changes in mitochondrial subcellular location and morphology accompany activation of the CO2 concentrating mechanism

Dynamic changes in intracellular ultrastructure can be critical for the ability of organisms to acclimate to environmental conditions. Microalgae, which are responsible for ~50% of global photosynthesis, compartmentalize their Ribulose 1,5 Bisphosphate Carboxylase/Oxygenase (Rubisco) into a specialized structure known as the pyrenoid when the cells experience limiting CO2 conditions; this compartmentalization is a component of the CO2 Concentrating Mechanism (CCM), which facilitates photosynthetic CO2 fixation as environmental levels of inorganic carbon (Ci) decline. Changes in the spatial distribution of mitochondria in green algae have also been observed under CO2 limitation, although a role for this reorganization in CCM function remains unclear. We used the green microalga Chlamydomonas reinhardtii to monitor changes in mitochondrial position and ultrastructure as cells transition between high CO2 and Low/Very Low CO2 (LC/VLC). Upon transferring cells to VLC, the mitochondria move from a central to a peripheral cell location and orient in parallel tubular arrays that extend along the cell's apico-basal axis. We show that these ultrastructural changes correlate with CCM induction and are regulated by the CCM master regulator CIA5. The apico-basal orientation of the mitochondrial membranes, but not the movement of the mitochondrion to the cell periphery, is dependent on microtubules and the MIRO1 protein, with the latter involved in membrane-microtubule interactions. Furthermore, blocking mitochondrial respiration in VLC-acclimated cells reduces the affinity of the cells for Ci. Overall, our results suggest that mitochondrial repositioning functions in integrating cellular architecture and energetics with CCM activities and invite further exploration of how intracellular architecture can impact fitness under dynamic environmental conditions.

Prion protein prevents the inhibition of large-conductance calcium-activated potassium channel by Tau peptide K18 oligomers

Alzheimer's disease (AD) is a tauopathy characterized by the deposition of amyloid aggregates of hyperphosphorylated Tau protein and amyloid-β peptide (Aβ) in the brain. Nevertheless, a soluble, oligomeric forms of Tau and Aβ are considered to be the most neurotoxic species responsible for neurodegenerative processes in AD. The mechanism of action of these oligomers remains largely unclear. Previously, we demonstrated the inhibition of the large-conductance calcium-activated potassium channel (BKCa) by Aβ. Therefore, in the present study we investigated the effect of Tau protein on the BKCa activity. Furthermore, since prion protein (PrP) interacts with Tau and the N-terminal fragment of PrP, called N1, can be neuroprotective in tauopathies, we checked whether N1 can also act at the level of BKCa channel.In the studies we used monomers, oligomers and amyloid fibrils of aggregation-prone Tau fragment, called K18, carrying tauopathy-associated mutation - deletion of Lys280 (K18Δ280). Additionally, to induce formation of neurotoxic oligomers, K18Δ280 was phosphorylated by protein kinase A (PKA). The activity of the plasma membrane BKCa of hippocampal neurons was recorded using single-channel patch-clamp technique in both inside-out and outside-out modes, exposing the cytosolic or extracellular surface of the membrane, respectively.In the outside-out mode - performing the extracellular application of the neurotoxic oligomers of phosphorylated K18Δ280, we observed a significant and concentration-dependent decrease in the probability of opening (Po) of BKCa. The Po of BKCa was fully recovered after washing the oligomers out. In the case of the inside-out patch-clamp configuration, we found that the Po of BKCa was not affected by the oligomers. In contrast to the oligomers, the monomers and amyloid fibrils of K18Δ280 had no effect on the channel activity, analyzed in inside-out as well as outside-out modes. Noteworthy, upon incubation with N1, the oligomers did not inhibit BKCa channel.The BKCa channel inhibition, dependent on the outside-out membrane orientation, implies specific interaction of the oligomers with the extracellular part of the channel. Moreover, our results suggest that N1 can convert the neurotoxic oligomers of Tau into a form which is not able to inhibit the channel, and indicate novel possible neuroprotective mechanism of PrP action in AD and other tauopathies.

Covalent Organic Framework Membranes with Regulated Orientation for Monovalent Cation Sieving.

Continuous covalent organic framework (COF) thin membranes have garnered broad concern over the past few years due to their merits of low energy requirements, operational simplicity, ecofriendliness, and high separation efficiency in the application process. This study marks the first instance of fabricating two distinct, self-supporting COF membranes from identical building blocks through solvent modulation. Notably, the precision of the COF membrane's separation capabilities is substantially enhanced by altering the pore alignment from a random to a vertical orientation. Within these confined channels, the membrane with vertically aligned pores and micron-scale stacking thickness demonstrates rapid and selective transportation of Li+ ions over Na+ and K+ ions, achieving Li+/K+ and Li+/Na+ selectivity ratios of 38.7 and 7.2, respectively. This research not only reveals regulated orientation and layer stacking in COF membranes via strategic solvent selection but also offers a potent approach for developing membranes specialized in Li+ ion separation.

Response of Staphylococcus aureus physiology and Agr quorum sensing to low-shear modeled microgravity.

Staphylococcus aureus is commonly isolated from astronauts returning from spaceflight. Previous analysis of omics data from S. aureus low Earth orbit cultures indicated significantly increased expression of the Agr quorum sensing system and its downstream targets in spaceflight samples compared to ground controls. In this current study, the rotary cell culture system (RCCS) was used to investigate the effect of low-shear modeled microgravity (LSMMG) on S. aureus physiology and Agr activity. When cultured in the same growth medium and temperature as the previous spaceflight experiment, S. aureus LSMMG cultures exhibited decreased agr expression and altered growth compared to normal gravity control cultures, which are typically oriented with oxygenation membrane on the bottom of the high aspect rotating vessel (HARV). When S. aureus was grown in an inverted gravity control orientation (oxygenation membrane on top of the HARV), reduced Agr activity was observed relative to both traditional control and LSMMG cultures, signifying that oxygen availability may affect the observed differences in Agr activity. Metabolite assays revealed increased lactate and decreased acetate excretion in both LSMMG and inverted control cultures. Secretomics analysis of LSMMG, control, and inverted control HARV culture supernatants corroborated these results, with inverted and LSMMG cultures exhibiting a decreased abundance of Agr-regulated virulence factors and an increased abundance of proteins expressed in low-oxygen conditions. Collectively, these studies suggest that the orientation of the HARV oxygenation membrane can affect S. aureus physiology and Agr quorum sensing in the RCCS, a variable that should be considered when interpreting data using this ground-based microgravity model.IMPORTANCES. aureus is commonly isolated from astronauts returning from spaceflight and from surfaces within human-inhabited closed environments such as spacecraft. Astronaut health and immune function are significantly altered in spaceflight. Therefore, elucidating the effects of microgravity on S. aureus physiology is critical for assessing its pathogenic potential during long-term human space habitation. These results also highlight the necessity of eliminating potential confounding factors when comparing simulated microgravity model data with actual spaceflight experiments.

Prediction of difficult round window visibility during cochlear implantation via a reformatted CT facial recess view: A retrospective study with surgical correlation.

Cochlear implant surgery is performed commonly through the facial recess via the round window (RW) approach. This study aims to evaluate the utility of reformatting the pre-operative CT temporal bone scan into a CT facial recess view in alerting surgeons to a potentially difficult surgery with poorly visualized round window. This is a retrospective study of 41 patients (43 ears), who had undergone cochlear implant surgery. Intraoperative findings of round window position relative to 2nd genu-mastoid portion of facial nerve, and round window membrane orientation were recorded by the surgeons. Pre-operative CTs were analyzed by two radiologists in axial and a reformatted facial recess plane that simulates the surgeon's view via the facial recess. Radiological assessment markers include the facial nerve-chorda tympani nerve width (FN-CTN) measured 1.2mm inferior to the exit point of the chorda tympani nerve into the tympanic cavity, round window position relative to 2nd genu-mastoid segment of the facial nerve and RW membrane's angle from the vertical axis. The best predictor for difficult round window intraoperative visibility is the RW position relative to the 2nd genu-mastoid segment of the facial nerve lying lateral to it on CT facial recess reformatted images. A RW that lies partially to completely posterior to the posterior border of the 2nd genu-mastoid segment of the facial nerve had up to 55.6% risk of encountering difficult access, while those positioned anterior to or partially anterior to the anterior edge of the 2nd genu-mastoid segment of the facial nerve had 0% risk of difficult access (p<0.05). There are substantial agreements in the intra-rater (Kappa=0.751, p<0.001) and inter-rater reliability (Kappa= 0.698, p<0.001). There is no significant association between surgical difficulty and facial nerve-chorda tympani distance or RW angle (p>0.05). Identification of round window positions in the reformatted CT facial recess view is a useful tool in predicting potentially difficult round window access in cochlear implant surgery. RW= Round window, FN-CTN= facial nerve to chorda tympani nerve width.

Electric field-driven assembly of (110) oriented metal–organic framework ZIF-8 monolayer with high hydrogen selectivity

Metal-organic framework (MOF) membranes have showed great potential in gas separation. To date, it is still a great challenge to precisely control the optimal orientation of MOF membranes with an enhanced gas separation performance. Herein, we prepared a compact and (110) oriented ZIF-8 membrane with a thickness of about 265 nm on graphite substrate within 1 h at room temperature via electric-field-driven strategy. Since the membrane is an intergrown one-crystal layer, grain boundary retardation of the flux is eliminated. Furthermore, due to the reduction of the transport pathways through the oriented structure, the (110) oriented ZIF-8 membrane exhibits a high gas separation performance. At 100 °C and 1 bar for the separation of equimolar binary gas mixtures H2/CO2, H2/CH4 and H2/C3H8, separation factors of 49.5, 90.9 and 121.8 can be obtained, with H2 permeance of about 1.7 × 10-7 mol·m−2·s−1·Pa−1. The developed strategy exhibits excellent reproducibility and scalability, and a large area ZIF-8 membrane can be consistently prepared on the graphite substrate, thus facilitating the preparation and application of the ZIF-8 membrane on a large scale. Further, the electric-field-driven strategy is also helpful for the preparation oriented ZIF-67 and ZIF-L membranes, confirming its versatility for the preparation of oriented MOF membranes.

A synthetic method to assay polycystin channel biophysics.

Ion channels are biological transistors that control ionic flux across cell membranes to regulate electrical transmission and signal transduction. They are found in all biological membranes and their conductive state kinetics are frequently disrupted in human diseases. Organelle ion channels are among the most resistant to functional and pharmacological interrogation. Traditional channel protein reconstitution methods rely upon exogenous expression and/or purification from endogenous cellular sources which are frequently contaminated by resident ionophores. Here we describe a fully synthetic method to assay functional properties of polycystin channels that natively traffic to primary cilia and endoplasmic reticulum organelles. Using this method, we characterize their oligomeric assembly, membrane integration, orientation and conductance while comparing these results to their endogenous channel properties. Outcomes define a novel synthetic approach that can be applied broadly to investigate channels resistant to biophysical analysis and pharmacological characterization.

Automated Piezo-Assisted Sperm Immobilization

Sperm immobilization is a crucial procedure in clinical cell surgery for infertility treatment. Current immobilization is implemented by tapping the sperm tail with a glass micropipette, but its effectiveness is restricted by sperm orientation and ineffective membrane ablation. Ineffective ablation leads to limited release of oocyte activating factors and lowers fertilization rate; and sperm swim in small angles relative to the micropipette tip cannot be tapped due to the risk of damaging the sperm’s genetic materials contained in the sperm head. This paper reports automated piezo-assisted sperm immobilization with enhanced efficacy of cell membrane ablation and sperm orientation control. The designed piezo drill consists of two orthogonal vibration modules to generate controlled micropipette vibration along axial and lateral axes. Through stiffness modeling, the flexure joints guide the motion of the central beam of each vibration module. To achieve sperm orientation control, whirl flow is induced by both axial and lateral vibration of the micropipette tip. To immobilize sperm, only micropipette’s axial vibration is generated to prevent lateral vibration from damaging sperm head. A visual servoing scheme is developed by decoupling sperm wiggling from positioning error for immobilization. Experimental results showed that sperm orientation control by the piezo drill achieved an error of 1.4 <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$^{\circ}$</tex-math> </inline-formula> and a time cost of 2.5 s. Visual servoing with sperm wiggling decoupling achieved a positioning error of 1.7 <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\mu$</tex-math> </inline-formula> m. Furthermore, the piezo-assisted sperm immobilization technique led to effective membrane ablation. With membrane-impermeable stains, it took 5.6 s for the immobilized sperm to be stained after piezo-assisted immobilization, significantly less than 49.2 s by conventional micropipette tapping. <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">Note to Practitioners</i> —This work tackled the challenge of ineffective membrane ablation and orientation limit in clinical cell surgeries. Conventional manual immobilization suffers from low membrane ablation efficacy, which leads to limited release of oocyte activating factors and lowers fertilization rate. Moreover, sperm swim in small angles relative to the micropipette tip cannot be tapped due to the risk of damaging the sperm’s genetic materials contained in the sperm head. In this paper, we propose automation techniques for effective membrane ablation and orientation control of sperm. A clinically compatible piezo drill is developed to generate controllable micropipette motion along both axial and lateral directions. The whirl flow generated by micropipette vibration is employed to rotate sperm, which greatly increased the number of available sperm for immobilization. A visual servoing controller is developed to keep the sperm at the center of field of view for immobilization by decoupling sperm wiggling from positioning error. The developed methods can be generalized to the manipulation of other types of cells. The piezo drill can be used for effective membrane ablation of oocyte, embryo, yeast cell and so on. The orientation control strategy leveraging piezo-induced whirl flow is applicable to non-contact rotation of a variety of microorganism.

Explicit expression for water permeation flux in forward osmosis desalination process

Forward osmosis (FO) is a membrane process of water separation and purification. FO uses the osmotic pressure difference across a semipermeable membrane. The effective osmotic pressure at the membrane–solution interface on both the feed and permeate sides of the membrane is the main driving force for the generation of the water flux. The major hindrance to the permeation of the water flux is the prevalence of concentration polarization on both sides of the membrane. Concentration polarization inhibits permeate flux by increasing the osmotic pressure at the membrane active layer interface on the feed side of the membrane. This work focused on the development of a mathematical model for water flux in the FO process. Combined film theory model and diffusion transport through the membrane were utilized. The effect of internal concentration polarization and external concentration polarization on the flux was studied. Both internal and external concentration polarization were taken into consideration in both membrane orientations, i.e., active layer facing the feed solution and active layer facing the draw solution. The obtained explicit expression for water permeation flux in forward osmosis desalination process shows excellent agreement with the literature data.

Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology

PIN proteins establish the auxin concentration gradient, which coordinates plant growth. PIN1-4 and 7 localized at the plasma membrane (PM) and facilitate polar auxin transport while the endoplasmic reticulum (ER) localized PIN5 and PIN8 maintain the intracellular auxin homeostasis. Although an antagonistic activity of PIN5 and PIN8 proteins in regulating the intracellular auxin homeostasis and other developmental events have been reported, the membrane topology of these proteins, which might be a basis for their antagonistic function, is poorly understood. In this study we optimized digitonin based PM-permeabilizing protocols coupled with immunocytochemistry labeling to map the membrane topology of PIN5 and PIN8 in Arabidopsis thaliana root cells. Our results indicate that, except for the similarities in the orientation of the N-terminus, PIN5 and PIN8 have an opposite orientation of the central hydrophilic loop and the C-terminus, as well as an unequal number of transmembrane domains (TMDs). PIN8 has ten TMDs with groups of five alpha-helices separated by the central hydrophilic loop (HL) residing in the ER lumen, and its N- and C-terminals are positioned in the cytoplasm. However, the topology of PIN5 comprises nine TMDs. Its N-terminal end and the central HL face the cytoplasm while its C-terminus resides in the ER lumen. Overall, this study shows that PIN5 and PIN8 proteins have a divergent membrane topology while introducing a toolkit of methods for studying membrane topology of integral proteins including those localized at the ER membrane.

Automated quantification of photoreceptor outer segments in developing and degenerating retinas on microscopy images across scales

The functionality of photoreceptors, rods, and cones is highly dependent on their outer segments (POS), a cellular compartment containing highly organized membranous structures that generate biochemical signals from incident light. While POS formation and degeneration are qualitatively assessed on microscopy images, reliable methodology for quantitative analyses is still limited. Here, we developed methods to quantify POS (QuaPOS) maturation and quality on retinal sections using automated image analyses. POS formation was examined during the development and in adulthood of wild-type mice via light microscopy (LM) and transmission electron microscopy (TEM). To quantify the number, size, shape, and fluorescence intensity of POS, retinal cryosections were immunostained for the cone POS marker S-opsin. Fluorescence images were used to train the robust classifier QuaPOS-LM based on supervised machine learning for automated image segmentation. Characteristic features of segmentation results were extracted to quantify the maturation of cone POS. Subsequently, this quantification method was applied to characterize POS degeneration in “cone photoreceptor function loss 1” mice. TEM images were used to establish the ultrastructural quantification method QuaPOS-TEM for the alignment of POS membranes. Images were analyzed using a custom-written MATLAB code to extract the orientation of membranes from the image gradient and their alignment (coherency). This analysis was used to quantify the POS morphology of wild-type and two inherited retinal degeneration (“retinal degeneration 19” and “rhodopsin knock-out”) mouse lines. Both automated analysis technologies provided robust characterization and quantification of POS based on LM or TEM images. Automated image segmentation by the classifier QuaPOS-LM and analysis of the orientation of membrane stacks by QuaPOS-TEM using fluorescent or TEM images allowed quantitative evaluation of POS formation and quality. The assessments showed an increase in POS number, volume, and membrane coherency during wild-type postnatal development, while a decrease in all three observables was detected in different retinal degeneration mouse models. All the code used for the presented analysis is open source, including example datasets to reproduce the findings. Hence, the QuaPOS quantification methods are useful for in-depth characterization of POS on retinal sections in developmental studies, for disease modeling, or after therapeutic interventions affecting photoreceptors.

Perfluorooctanoic acid-contaminated wastewater treatment by forward osmosis: Performance analysis

Perfluorooctanoic acid (PFOA) is a persistent compound, raising considerable global apprehension due to its resistance to breakdown and detrimental impacts on human health and aquatic environments. Pressure-driven membrane technologies treating PFAS-contaminated water are expensive and prone to fouling. This study presented a parametric investigation of the effectiveness of cellulose triacetate membrane in the forward osmosis (FO) membrane for removing PFOA from an aqueous solution. The study examined the influence of membrane orientation modes, feed pH, draw solution composition and concentration, and PFOA concentration on the performance of FO. The experimental results demonstrated that PFOA rejection was 99 % with MgCl2 and slightly >98 % with NaCl draw solutions due to the mechanism of PFOA binding to the membrane surface through Mg2+ ions. This finding highlights the crucial role of the draw solution's composition in PFOA treatment. Laboratory results revealed that membrane rejection of PFOA was 99 % at neutral and acidic pH levels but decreased to 95 % in an alkaline solution at pH 9. The decrease in membrane rejection is attributed to the dissociation of the membrane's functional groups, consequently causing pore swelling. The results were confirmed by calculating the average pore radius of the CTA membrane, which increased from 27.94 nm at pH 5 to 30.70 nm at pH 9. Also, variations in the PFOA concentration from 5 to 100 mg/L did not significantly impact the membrane rejection, indicating the process's capability to handle a wide range of PFOA concentrations. When seawater was the draw solution, the FO membrane rejected 99 % of PFOA concentrations ranging from 5 mg/L to 100 mg/L. The CTA FO treating PFOA-contaminated wastewater from soil remediation achieved a 90 % recovery rate and water flux recovery of 96.5 % after cleaning with DI water at 40 °C, followed by osmotic backwash. The results suggest the potential of using abundant and cost-effective natural solutions in the FO process, all without evident membrane fouling.

Different lateral packing stress in acyl chains alters KcsA orientation and structure in lipid membranes

The molecular structures of the various intrinsic lipids in membranes regulate lipid-protein interactions. These different lipid structures with unique volumes produce different lipid molecular packing stresses/lateral stresses in lipid membranes. Most studies examining lipid packing effects have used phosphatidylcholine and phosphatidylethanolamine (PE), which are the main phospholipids of eukaryotic cell membranes. In contrast, Gram-negative or Gram-positive bacterial membranes are composed primarily of phosphatidylglycerol (PG) and PE, and the physical and thermodynamic properties of each acyl chain in PG at the molecular level remain unresolved. In this study, we used 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (POPG, 16:0–18:1 PG) and 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (PAPG, 16:0–20:4 PG) to prepare lipid bilayers (liposome) with the rod-type fluorescence probe DPH. We measured the lipid packing conditions by determining the rotational freedom of DPH in POPG or PAPG bilayers. Furthermore, we investigated the effect of different monoacyl chains on a K+ channel (KcsA) structure when embedded in POPG or PAPG membranes. The results revealed that differences in the number of double bonds and carbon chain length in the monoacyl chain at sn-2 affected the physicochemical properties of the membrane and the structure and orientation of KcsA.

Synthesis optimization and separation mechanism of ZSM-5 zeolite membranes for pervaporation dehydration of organic solvents

Pervaporation (PV), as an energy-efficient mixture separation technology, plays an important role in the chemical industry. In this work, no organic templates were needed to produce high-performance ZSM-5 membranes with an extremely low Si/Al ratio of 3.3 on α-Al2O3 tubular supports using 100 nm nanoseeds. The effects of preparation parameters on the crystalline phase structures, micromorphologies, and PV separation performance of ZSM-5 membranes were comprehensively investigated. The results revealed that the Si/Al ratio of gels significantly affected both the Si/Al ratio and the crystal orientation of the final ZSM-5 membrane. The optimized ZSM-5 membrane with a thickness of 1.8 μm was utilized to dehydrate various organic solvents via PV, and the influence of the operating parameters on PV dehydration performance was evaluated and is described herein. Furthermore, the permeation behaviors of single gases and PV were examined using permeate molecules within a similar size range to reveal the PV mechanism of the ZSM-5 membrane. The results demonstrated that gas permeation followed Knudsen diffusion, while PV permeation was decreased with decreases in the affinity of molecules, revealing an adsorption-diffusion mechanism that dominated PV dehydration through the ZSM-5 membrane. Moreover, the as-synthesized ZSM-5 membrane had good water permselectivity for water/acetone (e.g., total flux = 1.03 kg/(m2 h), α = 307) and for water/isopropanol (e.g., total flux = 1.49 kg/(m2 h), α = 1070) mixtures compared with other membranes reviewed in the literature. The synthesized ZSM-5 membrane also exhibited excellent reproducibility, high stability, and attractive PV separation performance, demonstrating its significant potential application in the PV dehydration of organic solvents.

Synthesis and characterization of an innovative sodium alginate/polyvinyl alcohol bioartificial hydrogel for forward-osmosis desalination.

Recently, hydrogels have been widely applied as draw agents in forward osmosis (FO) desalination. This work aims to synthesize bioartificial hydrogel from a blend of sodium alginate (SA) and polyvinyl alcohol (PVA) using epichlorohydrin (ECH) as a crosslinker. Then this prepared hydrogel was applied as a draw agent with cellulose triacetate membrane in a batch (FO) cell. The effects of the PVA content in the polymer blend and the crosslinker dose on the hydrogel's swelling capacity were investigated to optimize the hydrogel's composition. Furthermore, the water flux and the reverse solute flux of the optimum SA/PVA hydrogel were evaluated in a batch (FO) unit under the effect of the hydrogel's particle size, feed solution (FS) temperature, FS concentration, and membrane orientation. Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD) and compression strength tests were used to characterize the prepared hydrogel. Results revealed that the equilibrium swelling ratio (%) of 5228 was achieved with a hydrogel that had 25% PVA and a crosslinking ratio of 0.8. FO experiments revealed that the maximum water flux of 0.845 LMH achieved, when distilled water was used as FS, average hydrogel's particle size was 60µm, and the FS temperature was 40°C.

Nonthermal concentration of skimmed goat milk by forward osmosis: The insights into process performance and product quality

Nonthermal processing technologies have been gained increasing attention in the dairy industry recently. In this study, the feasibility and effects of forward osmosis (FO) process as a nonthermal technique in the concentration of skimmed goat milk (SGM) were assessed and compared with thermal evaporation. The effects of operational conditions including membrane orientation, crossflow rate, concentration of draw solution (DS), and temperature of feed solution (FS) on process performance and concentration efficiency were explored. The total solids content of raw SGM can be increased from 9.07% to 18.78% through a single-stage FO process to reach a concentration factor of 2.0. Membrane fouling could be mitigated by improving the crossflow rate. Osmotic backwash proved to be the most effective cleaning approach to restore membrane performance with a flux recovery rate reaching 100%. The nonthermal FO process showed the superior performance to traditional thermal evaporation in retaining the natural characteristics of milk. Economic and energy evaluation indicated that such FO process with a pilot scale could be promising in the milk concentration at a low cost of $2.69/m3 with a low energy consumption of 4.80 kWh/m3. It could be an effective and promising concentration approach for producing concentrated milk with promoted quality.

Conserved allosteric perturbation of the GTPase domains by region 1 of Ras hypervariable regions

Ras proteins are important intracellular signaling hubs that can interact with numerous downstream effectors and upstream regulators through their GTPase domains (G-domains) anchored to plasma membranes by the C-terminal hypervariable regions (HVRs). The biological functions of Ras were proposed to be regulated at multiple levels including the intramolecular G-domain-HVR interactions, of which the exact mechanism and specificity are still controversial. Here, we demonstrate that the HVRs, instead of having direct contacts, can weakly perturb the G-domains via an allosteric interaction that is restricted to a ∼20 Å range and highly conserved in the tested Ras isoforms (HRas and KRas4B) and nucleotide-bound states. The origin of this allosteric perturbation has been localized to a short segment (residues 167–171) coinciding with region 1 of HVRs, which exhibits moderate to weak α-helical propensities. A charge-reversal mutation (E168K) of KRas4B in region 1, previously described in the Catalog of Somatic Mutations in Cancer database, was found to induce similar chemical shift perturbations as truncation of the HVR does. Further membrane paramagnetic relaxation enhancement (mPRE) data show that this region 1 mutation alters the membrane orientations of KRas4B and moderately increases the relative population of the signaling-compatible state.

Organization of Carotenoid Aggregates in Membranes Studied Selectively using Resonance Raman Optical Activity.

In living organisms, carotenoids are incorporated in biomembranes, remarkably modulating their mechanical characteristics, fluidity, and permeability. Significant resonance enhancement of Raman optical activity (ROA) signals of carotenoid chiral aggregates makes resonance ROA (RROA), a highly selective tool to study exclusively carotenoid assemblies in model membranes. Hence, RROA is combined with electronic circular dichroism (ECD), dynamic light scattering (DLS), molecular dynamics, and quantum-chemical calculations to shed new light on the carotenoid aggregation in dipalmitoylphosphatidylcholine (DPPC) liposomes. Using representative members of the carotenoid family: apolar α-carotene and more polar fucoxanthin and zeaxanthin, the authors demonstrate that the stability of carotenoid aggregates is directly linked with their orientation in membranes and the monomer structures inside the assemblies. In particular, polyene chain distortion of α-carotene molecules is an important feature of J-aggregates that show increased orientational freedom and stability inside liposomes compared to H-assemblies of more polar xanthophylls. In light of these results, RROA emerges as a new tool to study active compounds and drugs embedded in membranes.

Covalent Organic Network Membranes with Tunable Nanoarchitectonics from Macrocycle Building Blocks for Graded Molecular Sieving.

Traditional piperazine-based polyamide membranes usually suffer from the intrinsic trade-off relationship between selectivity and permeance. The development of macrocycle membranes with customized nanoscale pores is expected to address this challenge. Herein, we introduce 1,4-diazacyclohexane (2N), 1,4,7-triazacyclononane (3N), and 1,4,8,11-tetraazacyclotetradecane (4N) as molecular building blocks to construct the nanoarchitectonics of polyamide membranes prepared from interfacial polymerization (IP). The permeance of covalent organic network membranes follows the trend of 4N-TMC > 3N-TMC > 2N-TMC, while the molecular weight cutoff (MWCO) also follows the same trend of 4N-TMC > 3N-TMC > 2N-TMC, according to their nanopore size of the membranes. The microporosity, orientation, and surface chemistry of covalent organic network membranes can be rationally designed by macrocycle building units. The ordered nanoarchitectonics allows the membranes to attain an excellent performance in graded molecular sieving. Importantly, the novel covalent organic network membranes with tunable nanoarchitectonics prepared from macrocycle building units exhibited high water permeance (32.5 LMH/bar) and retained long-term stability after 100 h of test and bovine serum albumin fouling. These results reveal the enormous potential of 3N-TMC and 4N-TMC membranes in saline textile wastewater treatments and precise molecular sieving.