Pore Closure Research Articles

Gradient microstructures and mechanical properties of WAAMed and aged Al–Cu alloy after laser shock peening

The integration of wire-arc additive manufacture (WAAM) and laser shock peening (LSP) facilitated the study of the gradient microstructure in T6-treated WAAM Al–Cu alloy and the synergistic strengthening mechanisms involving the precipitation phase and LSP parameters. The results indicate that pore closures, increases in kernel average misorientation (KAM), and low angle grain boundary (LAGB) are positively associated with LSP energy density. Residual stress positively correlates with LSP energy density and inversely with spot diameter, where larger spots exacerbate non-uniformity. The values of dislocation density, microhardness and residual stress of the post-LSP specimen gradually increase with the increase of depth, and reach the maximum value at ~100 μm. As the depth continues to increase, the value begins to decrease gradually. Dislocation accumulation near grain boundaries, interacting with θ’-Al2Cu, contributes to strength enhancement. The main strengthening mechanisms enhanced by LSP involve creating a gradient microstructure through significant plastic deformation, pore closure, and the presence of high-density dislocations. Post-T6 treatment, θ’-Al2Cu significantly restricts dislocation slip, pins dislocations, and encourages the emergence of new dislocation sources, thereby enhancing the strengthening effect and stability induced by LSP.

Hybrid chitosan/HKUST-1 hydrogel with freezing and thawing modification as sustainable porous material for removal and selective separation of dye mixtures

A hybrid chitosan monolithic hydrogel (CS) was modified by incorporating with HKUST-1 and fabricated using an in-situ growth method. The resulting hydrogel CS/HKUST-1 was further modified to enhance the porous structure, in which the porous converted from close pores to interconnected porous structures. The chemical and physical characteristics of the modified hydrogel were examined using various characterization techniques like BET, SEM, EDX, FTIR, XRD, Zeta potential, and compression test to understand the effect of the modification. The outcomes displayed that this method provided homogeneous dispersing of HKUST-1 crystals within the chitosan hydrogel matrix, which can improve the material's specific surface area and adsorption capacity. The CS/HKUST-1 monolithic hybrid hydrogel's permeability is further enhanced by the freezing and thawing process and the produced hydrogel is called FCS/HKUST-1, which helps to open the hydrogel's pores and significantly increases its surface area. FCS/HKUST-1 hydrogels display higher mechanical properties without fracture after compression with elastic properties, whereas the CS/HKUST-1 hydrogel crashed at around 47 % compression strain. Consequently, the obtaining dual modified chitosan/ HKUST-1 hydrogel (FCS/HKUST-1) was evaluated as an adsorbent to eliminate Eosin Y (EY) dye from its aqueous solution and the adsorption mechanism was proposed as multilayer adsorption depending on the Freundlich isotherm model. The modified monolithic hydrogel displayed an excellent maximum adsorption capacity with EY dye about 625 mg·g−1. The FCS/HKUST-1 hydrogel showed extremely selective removal for anionic dye higher than cationic dye, which allows facile separation of the dyes with different charges in a mixture solution. The outcomes also demonstrate that the modified hydrogel would be an eco-friendly material, which shows excellent recyclability, indicating that this material considerable promising adsorbent in water treatment.

Fighting for Survival at the Stomatal Gate.

Stomata serve as the battleground between plants and plant pathogens. Plants can perceive pathogens, inducing closure of the stomatal pore, while pathogens can overcome this immune response with their phytotoxins and elicitors. In this review, we summarize new discoveries in stomata-pathogen interactions. Recent studies have shown that stomatal movement continues to occur in a close-open-close-open pattern during bacterium infection, bringing a new understanding of stomatal immunity. Furthermore, the canonical pattern-triggered immunity pathway and ion channel activities seem to be common to plant-pathogen interactions outside of the well-studied Arabidopsis-Pseudomonas pathosystem. These developments can be useful to aid in the goal of crop improvement. New technologies to study intact leaves and advances in available omics data sets provide new methods for understanding the fight at the stomatal gate. Future studies should aim to further investigate the defense-growth trade-off in relation to stomatal immunity, as little is known at this time.

Experimental and molecular dynamics multiscale study on sintering and interfacial reaction of Fe2O3/CaO particle system

High-temperature sintering and solid-state reaction within the intricate Fe2O3/CaO system are pivotal factors influencing material characteristics. The current work focused on the sintering and reaction behavior of Fe2O3/CaO particles at high temperatures with a multiscale study of in-situ high-temperature stage microscopy (HTSM) and molecular dynamics (MD). The laws governing atomic migration and the evolution of crystal structure during the sintering process were simulated at the nanoscale. The results elucidated that a heightened heating rate fosters particle shrinkage and concomitant reduction in surface area. The sintering activation energy (Esin) exhibited an increment proportional to the linear shrinkage of the particles. Furthermore, an augmented potential barrier in advanced stages impeded subsequent sintering behavior. Augmented temperature promoted the particle shrinkage and sintering neck formation, culminating in pore closure and a diminution in the free surface. However, this promotional effect attenuated with escalating temperatures. Particle shrinkage and sintering neck formation are contingent upon atomic migration and diffusion, with the growth of sintering necks primarily reliant on atom diffusion along the surface of the sintering necks. The atomic diffusion activation energy was determined to be 49.5 kJ/mol within the temperature range of 1073–1473 K. Within the Fe2O3/CaO system, the preferential generation of Ca2Fe2O5 (C2F) over CaFe2O4 (CF) was found, leading to a decrease in the atomic diffusion barrier and an enhancement of the diffusivity, with the latter dominating in later stages. The highest mobility of O2–, Fe3+, and Ca2+ was discerned in CF, and the sintering shrinkage of the particles predominantly hinges on the generation of CF.

Activation of Polycystin-1 Signaling by Binding of Stalk-derived Peptide Agonists.

Polycystin-1 (PC1) is the membrane protein product of the PKD1 gene whose mutation is responsible for 85% of the cases of autosomal dominant polycystic kidney disease (ADPKD). ADPKD is primarily characterized by the formation of renal cysts and potential kidney failure. PC1 is an atypical G protein-coupled receptor (GPCR) consisting of 11 transmembrane helices and an autocatalytic GAIN domain that cleaves PC1 into extracellular N-terminal (NTF) and membrane-embedded C-terminal (CTF) fragments. Recently, signaling activation of the PC1 CTF was shown to be regulated by a stalk tethered agonist (TA), a distinct mechanism observed in the adhesion GPCR family. A novel allosteric activation pathway was elucidated for the PC1 CTF through a combination of Gaussian accelerated molecular dynamics (GaMD), mutagenesis and cellular signaling experiments. Here, we show that synthetic, soluble peptides with 7 to 21 residues derived from the stalk TA, in particular, peptides including the first 9 residues (p9), 17 residues (p17) and 21 residues (p21) exhibited the ability to re-activate signaling by a stalkless PC1 CTF mutant in cellular assays. To reveal molecular mechanisms of stalk peptide-mediated signaling activation, we have applied a novel Peptide GaMD (Pep-GaMD) algorithm to elucidate binding conformations of selected stalk peptide agonists p9, p17 and p21 to the stalkless PC1 CTF. The simulations revealed multiple specific binding regions of the stalk peptide agonists to the PC1 protein including an "intermediate" bound yet inactive state. Our Pep-GaMD simulation findings were consistent with the cellular assay experimental data. Binding of peptide agonists to the TOP domain of PC1 induced close TOP-putative pore loop interactions, a characteristic feature of the PC1 CTF signaling activation mechanism. Using sequence covariation analysis of PC1 homologs, we further showed that the peptide binding regions were consistent with covarying residue pairs identified between the TOP domain and the stalk TA. Therefore, structural dynamic insights into the mechanisms of PC1 activation by stalk-derived peptide agonists have enabled an in-depth understanding of PC1 signaling. They will form a foundation for development of PC1 as a therapeutic target for the treatment of ADPKD.

Depth dewatering of kaolinite cake by SDBS/PEI synergy and a mesoscopic mechanism study

The objective of this study was to elucidate the synergistic dewatering mechanism of mixed sodium dodecylbenzene sulfonate (SDBS) and polyethyleneimine (PEI) on kaolinite filter cakes and to test the dewatering effectiveness of a custom-built pressure dewatering device. The dewatering experiments showed that while the reagent dose was reduced by 50%, the dewatering rate decreased to 19.78%, surpassing the optimal dose of a single reagent. During the natural compression stage of the filter cake, the deformation reached its maximum under the same pressure difference. During the transitional period, the closure of the filter cake pores occurred at a slower rate, facilitating better moisture drainage. Under strong compression, the ultimate deformation reached its maximum. PEI acted as a bridging agent for kaolin flocculation and served as a medium for SDBS on the kaolin surface. A small amount of PEI reduced the electrostatic repulsion of SDBS, increasing its adsorption capacity. PEI adsorbed on the kaolin surface through electrostatic and hydrogen bonding, while SDBS could adsorb onto the PEI reagent layer through electrostatic and polar interactions, exposing its alkyl chains on the surface and thereby facilitating flocculation and generating a hydrophobic reagent layer. This study could provide a new method and insights for the efficient pressurized dewatering of kaolin.

Experimental study on the dynamic evolution of rock pore structure in real-time monitoring based on nuclear magnetic resonance

The rock structure determines the stability and reliability of underground engineering. Great achievements have been made in the study of rock structural damage, but there are few studies on real-time monitoring of rock structural changes under different increasing and unloading decreasing pressures. In this paper, the dynamic evolution rule of pore structure in rock under the combined action of water pressure and different confining pressure of increasing and decreasing is monitored by NMR technique. The porosity and permeability of rock under different confining pressures are consistent. The increase of confining pressure leads to the closure of pores, which leads to the decrease of porosity and permeability, but the amplitude and time of change are different. During the process of decreasing confining pressure, the T2 spectral curve, porosity and permeability of the rock did not fully recover due to the reduction of confining pressure, indicating that plastic deformation occurs inside the rock at this time. The mathematical relationship between porosity and permeability is established, and the permeability of rock is not only related to the size of porosity, but also closely related to the pore structure of rock.

A novel hydro-elastoplastic constitutive model incorporating hydrostatic damage for predicting high-pressure performance of concrete under blast loading

The evolution of damage and failure modes of concrete under blast or impact loads are the outcomes of stress wave propagation. The stress state at a point within the concrete changes instantaneously with time and space. Establishing a constitutive model that can cover the nonlinear behavior of materials under complex loading paths and histories is an extremely challenging task. The main difficulty lies in the construction of the damage model, which has not yet been fully resolved in commonly used concrete models. This paper, based on the mechanisms of damage and failure of concrete under hydrostatic pressure, constructs a state equation that includes the description of crushing and compaction damage due to the collapse and closure of pores, as well as their interactive model, negative pressure degradation caused by hydrostatic pressure, and nonlinear volumetric behavior under loading/unloading/reloading. By introducing independent shear and tensile deviatoric damage scaling functions and a damage scaling function under isotropic tension, the coupling relationship between deviatoric and volumetric damage is considered. Corresponding strength models and element erosion criteria are developed based on the new state equation and damage models. Then, single-element numerical experiments of unconstrained uniaxial compression and tension, triaxial compression under different confining pressures, monotonic and cyclic hydrostatic compression, and isotropic tension were conducted to verify the predictive accuracy of the proposed model under single loading paths. Finally, numerical experiments of contact explosion on plain concrete thick and thin targets were carried out using the proposed model, revealing the propagation laws and failure processes of the loading compression wave, unloading tensile wave, and reflected tensile wave within the concrete target. The predicted final failure modes are consistent with the experimental results.

Mechanical Strain Induces and Increases Vesicular Release Monitored by Microfabricated Stretchable Electrodes

AbstractExocytosis involving the fusion of intracellular vesicles with cell membrane, is thought to be modulated by the mechanical cues in the microenvironment. Single‐cell electrochemistry can offer unique information about the quantification and kinetics of exocytotic events; however, the effects of mechanical force on vesicular release have been poorly explored. Herein, we developed a stretchable microelectrode with excellent electrochemical stability under mechanical deformation by microfabrication of functionalized poly(3,4‐ethylenedioxythiophene) conductive ink, which achieved real‐time quantitation of strain‐induced vesicular exocytosis from a single cell for the first time. We found that mechanical strain could cause calcium influx via the activation of Piezo1 channels in chromaffin cell, initiating the vesicular exocytosis process. Interestingly, mechanical strain increases the amount of catecholamines released by accelerating the opening and prolonging the closing of fusion pore during exocytosis. This work is expected to provide revealing insights into the regulatory effects of mechanical stimuli on vesicular exocytosis.

Mechanical Strain Induces and Increases Vesicular Release Monitored by Microfabricated Stretchable Electrodes.

Exocytosis involving the fusion of intracellular vesicles with cell membrane, is thought to be modulated by the mechanical cues in the microenvironment. Single-cell electrochemistry can offer unique information about the quantification and kinetics of exocytotic events; however, the effects of mechanical force on vesicular release have been poorly explored. Herein, we developed a stretchable microelectrode with excellent electrochemical stability under mechanical deformation by microfabrication of functionalized poly(3,4-ethylenedioxythiophene) conductive ink, which achieved real-time quantitation of strain-induced vesicular exocytosis from a single cell for the first time. We found that mechanical strain could cause calcium influx via the activation of Piezo1 channels in chromaffin cell, initiating the vesicular exocytosis process. Interestingly, mechanical strain increases the amount of catecholamines released by accelerating the opening and prolonging the closing of fusion pore during exocytosis. This work is expected to provide revealing insights into the regulatory effects of mechanical stimuli on vesicular exocytosis.

Clathrin mediates membrane fission and budding by constricting membrane pores

Membrane budding, which underlies fundamental processes like endocytosis, intracellular trafficking, and viral infection, is thought to involve membrane coat-forming proteins, including the most observed clathrin, to form Ω-shape profiles and helix-forming proteins like dynamin to constrict Ω-profiles’ pores and thus mediate fission. Challenging this fundamental concept, we report that polymerized clathrin is required for Ω-profiles’ pore closure and that clathrin around Ω-profiles’ base/pore region mediates pore constriction/closure in neuroendocrine chromaffin cells. Mathematical modeling suggests that clathrin polymerization at Ω-profiles’ base/pore region generates forces from its intrinsically curved shape to constrict/close the pore. This new fission function may exert broader impacts than clathrin’s well-known coat-forming function during clathrin (coat)-dependent endocytosis, because it underlies not only clathrin (coat)-dependent endocytosis, but also diverse endocytic modes, including ultrafast, fast, slow, bulk, and overshoot endocytosis previously considered clathrin (coat)-independent in chromaffin cells. It mediates kiss-and-run fusion (fusion pore closure) previously considered bona fide clathrin-independent, and limits the vesicular content release rate. Furthermore, analogous to results in chromaffin cells, we found that clathrin is essential for fast and slow endocytosis at hippocampal synapses where clathrin was previously considered dispensable, suggesting clathrin in mediating synaptic vesicle endocytosis and fission. These results suggest that clathrin and likely other intrinsically curved coat proteins are a new class of fission proteins underlying vesicle budding and fusion. The half-a-century concept and studies that attribute vesicle-coat contents’ function to Ω-profile formation and classify budding as coat-protein (e.g., clathrin)-dependent or -independent may need to be re-defined and re-examined by considering clathrin’s pivotal role in pore constriction/closure.

Transient pores in hemifusion diaphragms

Exchange of material across two membranes, as in the case of synaptic neurotransmitter release from a vesicle, involves the formation and poration of a hemifusion diaphragm (HD). The nontrivial geometry of the HD leads to environment-dependent control, regarding the stability and dynamics of the pores required for this kind of exocytosis. This work combines particle simulations, field-based calculations, and phenomenological modeling to explore the factors influencing the stability, dynamics, and possible control mechanisms of pores in HDs. We find that pores preferentially form at the HD rim, and that their stability is sensitive to a number of factors, including the three line tensions, membrane tension, HD size, and the ability of lipids to “flip-flop” across leaflets. Along with a detailed analysis of these factors, we discuss ways that vesicles or cells may use them to open and close pores and thereby quickly and efficiently transport material.

New insights into the domain of unknown function (DUF) of EccC5, the pivotal ATPase providing the secretion driving force to the ESX-5 secretion system.

Type VII secretion (T7S) systems, also referred to as ESAT-6 secretion (ESX) systems, are molecular machines that have gained great attention due to their implications in cell homeostasis and in host-pathogen interactions in mycobacteria. The latter include important human pathogens such as Mycobacterium tuberculosis (Mtb), the etiological cause of human tuberculosis, which constitutes a pandemic accounting for more than one million deaths every year. The ESX-5 system is exclusively found in slow-growing pathogenic mycobacteria, where it mediates the secretion of a large family of virulence factors: the PE and PPE proteins. The secretion driving force is provided by EccC5, a multidomain ATPase that operates using four globular cytosolic domains: an N-terminal domain of unknown function (EccC5DUF) and three FtsK/SpoIIIE ATPase domains. Recent structural and functional studies of ESX-3 and ESX-5 systems have revealed EccCDUF to be an ATPase-like fold domain with potential ATPase activity, the functionality of which is essential for secretion. Here, the crystal structure of the MtbEccC5DUF domain is reported at 2.05 Å resolution, which reveals a nucleotide-free structure with degenerated cis-acting and trans-acting elements involved in ATP binding and hydrolysis. This crystallographic study, together with a biophysical assessment of the interaction of MtbEccC5DUF with ATP/Mg2+, supports the absence of ATPase activity proposed for this domain. It is shown that this degeneration is also present in DUF domains from other ESX and ESX-like systems, which are likely to exhibit poor or null ATPase activity. Moreover, based on an in silico model of the N-terminal region of MtbEccC5DUF, it is hypothesized that MtbEccC5DUF is a degenerated ATPase domain that may have retained the ability to hexamerize. These observations draw attention to DUF domains as structural elements with potential implications in the opening and closure of the membrane pore during the secretion process via their involvement in inter-protomer interactions.

Architecture and functions of stomatal cell walls in eudicots and grasses.

Like all plant cells, the guard cells of stomatal complexes are encased in cell walls that are composed of diverse, interacting networks of polysaccharide polymers. The properties of these cell walls underpin the dynamic deformations that occur in guard cells as they expand and contract to drive the opening and closing of the stomatal pore, the regulation of which is crucial for photosynthesis and water transport in plants. Our understanding of how cell wall mechanics are influenced by the nanoscale assembly of cell wall polymers in guard cell walls, how this architecture changes over stomatal development, maturation and ageing and how the cell walls of stomatal guard cells might be tuned to optimize stomatal responses to dynamic environmental stimuli is still in its infancy. In this review, we discuss advances in our ability to probe experimentally and to model the structure and dynamics of guard cell walls quantitatively across a range of plant species, highlighting new ideas and exciting opportunities for further research into these actively moving plant cells.

Investigation of pore structure evolution and damage characteristics of high temperature rocks subjected to liquid nitrogen cooling shock

Liquid nitrogen (LN2) can be utilized in the development of enhanced geothermal systems, as well as for deep/ultra-deep hydrocarbon reservoir stimulation, fire suppression, and other high-temperature geological projects. It is a crucial issue in the utilization of LN2 to investigate the pore structure evolution, permeability, and damage characteristics of high-temperature rocks under the influence of LN2 cooling shock. These rocks were first slowly heated to 150∼600°C and held for 2 h, followed by LN2 or natural cooling. The evolution of pore volume in high-temperature rocks affected by liquid nitrogen cooling was quantified. T2 cutoff values were determined through centrifugal tests, while the contents of irreducible and mobile fluids were estimated. Based on the aforementioned analysis as well as changes in irreducible fluid saturation, pore throat, tortuosity, and permeability, this study examines the closure and development of pores along with permeability behavior. The findings suggest that, despite a more pronounced decrease in porosity at lower heating temperatures, LN2 cooling specimens exhibit superior pore connectivity and permeability compared to those cooled naturally. LN2 stimulation not only induces crack initiation and propagation but also results in further cooling induced densification based on heating densification. 225°C is considered to be the optimal temperature for cooling contraction induced densification in this study. At higher heating temperatures, the damage to rock cooled with LN2 is more severe than that of naturally cooled. This results in a greater increase in porosity, movable fluid content and proportion, and permeability of LN2 cooled specimens compared to naturally cooled specimens. The damage mechanism can be better understood by the constructed damage model that coordinates the pore increase/decrease and mutual pore transformation from the perspective of pore evolution.

Moisture barriers used in firefighters' protective clothing: Effect of accelerated thermal aging on their mechanical and barrier performance

AbstractMoisture barriers are an essential component in firefighters' protective clothing as they allow water vapor to exit and prevent liquids from entering into the garment. However, they may be sensitive to thermal aging. This study investigates the effect of accelerated thermal aging at temperatures between 95 and 240°C on three different moisture barriers used in firefighters' protective clothing. Variations in their tear force, water vapor transmission rate (WVTR), and apparent contact angle were observed after thermal aging. These variations were influenced by factors such as the base fabric structure, fiber content, adhesive characteristics, and presence of an additional flame‐resistant polyurethane coating on top of the expanded polytetrafluoroethylene membrane. Adhesive degradation and delamination were observed in one moisture barrier, whilst different extents of base fabric degradation occurred in all three of them, leading to tear force reduction. Depending on the moisture barrier and aging temperature, the membranes experienced crack formation or pore closure, which affected the WVTR. The water‐repellent finish on the fabric side degraded in two moisture barriers but was not affected in the other. These findings highlight the importance of considering the constitutive materials and the structure of moisture barriers to understand their aging behavior and improve firefighters' safety.

Exploring Carbonization Temperature to Create Closed Pores for Hard Carbon as High-Performance Sodium-Ion Battery Anodes.

Biomass-derived porous carbon materials are meaningful to employ as a hard carbon precursor for anode materials of sodium-ion batteries (SIBs) from a sustainability perspective. Here, a straightforward approach is proposed to develop rich closed pores in pinenut-derived carbon, with the aim of improving Na+ plateau storage by adjusting the pyrolysis temperature. The optimized sample, namely the pinenut-derived carbon at 1300 °C, demonstrates remarkable reversible specific capacity of 278 mAh g-1, along with a high initial Coulomb efficiency of 85% and robust cycling stability (with a capacity retention of 89% after 800 cycles at 0.2 A g-1). In situ and ex situ analyses unveil that the developed closed pores play a significant role in enhancing the plateau capacity, providing compelling evidence for the "adsorption-filling" mechanism. Moreover, the corresponding full-cell achieves a high energy density of 245.7Wh kg-1 (based on the total weight of both electrode active materials) and exhibits outstanding rate capability (191.4 mAh g-1 at 3 A g-1).

A comprehensive study on the effects of surface post-processing on fatigue performance of additively manufactured AlSi10Mg: An augmented machine learning perspective on experimental observations

In this study, the efficacy of 21 distinct surface post-processing methods on the fatigue behavior of an additively manufactured (AM) material was investigated. Various treatments, encompassing single-step processes like sand blasting (SB), conventional shot peening (CSP), severe shot peening (SSP), gradient severe shot peening (GSSP), ultrasonic shot peening (USP), severe vibratory peening (SVP), ultrasonic nanocrystal surface modification (UNSM), laser shock peening (LSP), laser polishing (LP), tumble finishing (TF), chemical polishing (CP), electro-chemical polishing (ECP), and machining (M), as well as hybrid treatments, were systematically investigated for their impact on the fatigue behavior of hourglass laser powder based fused (LB-PBF) AlSi10Mg specimens. A comprehensive series of experimental tests were carried out, covering surface texture analyses, microstructural characterizations, porosity measurements, hardness, residual stresses (RS), monotonic tensile tests, and rotating bending fatigue tests at four stress levels. Following the experimental investigations, the acquired data were leveraged to develop a machine learning (ML)-based model to correlate the fatigue life with the influencing factors and conduct parametric and sensitivity analyses. The model utilized a deep learning approach to predict the fatigue response of LB-PBF AlSi10Mg, incorporating various artificial neural networks (ANNs) and considering input parameters such as yield strength, ultimate tensile strength, elongation to fracture, porosity, depth of pore closure, surface hardness, depth of hardness variation, surface RS, maximum compressive residual stresses (CRS), depth of the CRS field, top surface grain size, surface layer mean grain size, surface roughness, and stress amplitude. Depending on the life regime, the factors influencing fatigue behavior can vary significantly. In higher stresses, dominated by plastic strains, surface residual stress emerges as the primary factor. Conversely, in lower stresses, dominated by elastic strains, the significance of surface roughness becomes more pronounced, followed by surface residual stress, surface hardness and other factors. These findings underscore the contextual importance of different influencing factors for enhancing fatigue resistance based on varying loading conditions.

A novel RyR2 mutation associated with co-morbid catecholaminergic polymorphic ventricular tachycardia (CPVT) and benign epilepsy with centrotemporal spikes (BECTS)

In this case report, we describe a 14-year-old patient with a novel RyR2 gene mutation (c.6577G > T/p.Val2193Leu), identified through a comprehensive review of medical history, examination findings, and follow-up data. The pathogenic potential of this mutation, which results in the loss of some interatomic forces and compromises the closure of the RyR2 protein pore leading to calcium leakage, was analyzed using the I-TASSER Suite to predict the structural changes in the protein. This mutation manifested clinically as co-morbid catecholaminergic polymorphic ventricular tachycardia (CPVT) and benign epilepsy with centrotemporal spikes (BECTS), a combination not previously documented in the same patient. While seizures were successfully managed with levetiracetam, the patient's exercise-induced syncope episodes could not be controlled with metoprolol, highlighting the complexity and challenge in managing CPVT associated with this novel RyR2 variation.

Dual enzyme compartmentalization in a pH-responsive membrane: A way to tune enzymatic reactions in biocatalytic membranes

In this work, the use of a pH-responsive isoporous membrane allowed the compartmentalization of two different enzymes, one on the surface and the other in the pores of the membrane. This intensified and compartmentalized enzyme-loaded membrane system allowed the two enzymatic reactions to be modulated based on a pH shift. The chosen enzymes are pectinase and β-glucosidase, enzymes commonly used in biorefinery, whose optimum pH (3 and 6.5, respectively) is in the range where membrane pore closing (pH < 4) and opening is observed. Pectinase is covalently bound to the membrane surface at pH 3 while the pores are closed and is used to control the membrane fouling due to pectin accumulation during plant material processing. β-glucosidase is immobilized in the membrane pores at pH 6.5 (open pores) and is used to catalyze the reaction of oleuropein into the phytotherapeutic oleuropein aglycone. The different localization of the two enzymes was demonstrated by AFM, immunolocalization and SEM, confocal microscopy and XPS. In the open pore configuration, a very high conversion of pectin and oleuropein is simultaneously obtained (93% and 95%, in 165 h, respectively); whereas, in the closed pore configuration, an increased and exclusive conversion (95% in 24 h) of pectin was obtained. This innovative strategy of artificial ordered compartmentalization allows tuning the enzymatic reactions based on pH, preserving labile biomolecules (e.g.β-glucosidase) from deactivation at acidic pH and cleaning the membrane avoiding aggressive solvents. This makes it possible to increase the stability of enzymes, which is an obstacle to the development of biocatalytic membrane reactors, and proposes an unconventional and efficient way to synchronize the action of complementary enzymes in a case, or to selectively enhance the action of one of them by simply shifting the pH.