Search Results (158)

The present review summarizes a vast body of literature on the subject of Membrane Distillation (MD), with a special emphasis on the existing results and correlations for the transmembrane transport of heat and mass. The issue of saltwater physical properties is also discussed in depth, whereas the advective transport of heat and salt concentration in the feed and permeate compartments is only briefly mentioned but is beyond the scope of this review. The paper does not aim to provide a complete treatment of the subject of MD, which can be found in other publications. Rather, it suggests the data and correlations most suitable for the range of operating conditions typically expected in actual units implementing Direct Contact Membrane Distillation (DCMD), including hollow fiber designs, with a view to assist model development. The focus is on MD for water desalination, although some results may apply well to other fields. Full article

Cystic fibrosis (CF) is caused by loss-of-function variants in the cystic fibrosis transmembrane conductance regulator (CFTR) chloride and bicarbonate channel and affects multiple organs, with pancreatic involvement showing very high penetrance. In pancreatic ducts, CFTR drives secretion of alkaline, bicarbonate-rich fluid that maintains intraductal patency, neutralises gastric acid and permits safe delivery of digestive enzymes. Selective impairment of CFTR-dependent bicarbonate transport, even in the presence of residual chloride conductance, is strongly associated with exocrine pancreatic insufficiency, recurrent pancreatitis and cystic-fibrosis-related diabetes. These clinical manifestations are captured by pharmacodynamic anchors such as faecal elastase-1, steatorrhoea, pancreatitis burden and glycaemic control, providing clinically meaningful benchmarks for CFTR-targeted therapies. In this review, we summarise the principal mechanisms underlying pancreatic pathophysiology and the current approaches to clinical management. We then examine in vitro pancreatic duct models that are used to evaluate small molecules and emerging therapeutics targeting CFTR. These experimental systems include native tissue, primary cultures, organoids, co-cultures and microfluidic devices, each of which has its own advantages and limitations. Intact micro-perfused ducts provide the physiological benchmark for studying luminal pH control and bicarbonate (HCO₃⁻) secretion. Primary pancreatic duct epithelial cells (PDECs) and pancreatic ductal organoids (PDO) preserve ductal identity, patient-specific genotype and key regulatory networks. Immortalised ductal cell lines grown on permeable supports enable scalable screening and structure activity analyses. Co-culture models and organ-on-chip devices incorporate inflammatory, stromal and endocrine components together with flow and shear and provide system-level readouts, including duct-islet communication. Across this complementary toolkit, we prioritise bicarbonate-relevant endpoints, including luminal and intracellular pH and direct measures of HCO₃⁻ flux, to improve alignment between in vitro pharmacology and clinical pancreatic outcomes. The systematic use of complementary models should facilitate the discovery of next-generation CFTR modulators and adjunctive strategies with the greatest potential to protect both exocrine and endocrine pancreatic function in people with CF. Full article

Background: The ACE Y215C mutation is a common, functionally damaging missense variant (~1.5% allele frequency) associated with reduced plasma ACE levels and increased Alzheimer’s disease (AD) risk. In CHO and HEK cell models, this mutation caused a ~3–6-fold decrease in ACE surface expression, soluble ACE levels, and ACE enzymatic activity compared to those of wild-type ACE. Methods: Circulating ACE levels and activity were measured in EDTA plasma obtained from 84 carriers of the ACE Y215C mutation using a set of mAbs to the ACE. The mAbs 5B3/1G12 binding ratio was revealed as a sensitive marker for the circulating Y215C ACE mutant. Whole-exome and whole-genome sequencing (WES/WGS) were performed to identify genetic variants potentially modifying circulating ACE levels. In parallel, published sequencing and proteomic data from 35,559 Icelanders participants were analyzed to identify genes influencing ACE shedding. Sequence comparison was performed between carriers with elevated and reduced ACE concentrations to identify the potential protective variants that may compensate for decreased ACE levels due to the Y215C mutation itself. Results: Most carriers of the Y215C ACE mutation demonstrated significantly decreased ACE levels (median is 62% of control ACE levels). However, substantial inter-individual variability was observed in plasma ACE activity among carriers. Comparative sequencing analysis revealed 9648 variants unique to individuals with elevated ACE, mapping to 5779 protein-coding genes and enriched for pathways related to intracellular and transmembrane transport. Conclusions: The presence of the damaging ACE mutation Y215C does not invariably result in low plasma ACE or, likely, elevated AD risk. Therefore, combined blood ACE phenotyping and whole-exome sequencing are recommended to more accurately assess ACE-related AD susceptibility in mutation carriers. Full article

Background: The profound heterogeneity of glioblastoma and the often-limited efficacy of conventional treatments, including arsenic trioxide (ATO), underscore the urgent and critical demand for innovative combination strategies specifically designed to overcome treatment resistance. Methods: We evaluated the therapeutic effects of ATO as a single agent and in combination with the MNK1 inhibitor AUM001 across patient-derived xenograft (PDX) models and investigated molecular determinants of sensitivity and synergy. Our results demonstrated that GBM models resistant to ATO, particularly those of the mesenchymal subtype, are more likely to show synergistic cytotoxicity when AUM001 is added. The combination significantly reduces the frequency of glioblastoma stem cells (GSCs) compared to either drug alone, especially in ATO-resistant models. Results: These observations suggest that targeting the MNK1 pathway in conjunction with ATO is a promising strategy to specifically eradicate GSCs, which are major drivers of GBM recurrence and therapeutic failure. Transcriptomic analyses revealed that ATO sensitivity correlated with activated translation-related pathways and cell cycle processes, while synergistic responses to the combination were driven by distinct molecular signatures in different GBM subtypes. Overall, synergistic response to the combination therapy is more associated with cellular organization, amino acid transmembrane transporter activity, ion channels, extracellular matrix organization and collagen formation. Conclusions: Our findings highlight that specific molecular pathways and their activities, including those involving translation, cell cycle and ion transport, appear to modulate the synergistic efficacy of the ATO and AUM001 combination, thereby offering potential biomarkers for improved patient stratification in future GBM clinical trials of such ATO-based treatments. Full article

This study aimed to characterize the transcriptional response of Porphyromonas gingivalis biofilms to treatment with xanthohumol and curcumin. A validated dynamic in vitro biofilm model, based on microbial growth under flow and shear conditions resembling the oral cavity, was used to develop mature biofilms of P. gingivalis on sterile ceramic calcium hydroxyapatite discs. Transcriptional profiles of biofilms, treated and untreated with both extracts, were obtained through RNA-Sequencing (RNA-Seq). The biofilm development and the lack of phenotypic effects from sublethal concentrations of xanthohumol and curcumin were confirmed via Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM). Reverse transcription quantitative PCR (RT-qPCR) was employed to verify differentially expressed genes identified by RNA-Seq. Xanthohumol and curcumin caused extensive reprogramming of P. gingivalis biofilm gene expression. Out of 1,973 genes, xanthohumol activated 173 and repressed 286, whereas curcumin activated 170 and repressed 163. These changes affected genes involved in membrane integrity, oxidative stress, transmembrane transport, and virulence, suggesting a mechanism of action that involves membrane disruption. Full article

Skin hydration is fundamental for maintaining epidermal barrier integrity and overall skin homeostasis. Beyond traditional moisturizing agents, recent research has highlighted the role of aquaporins (AQPs), transmembrane water channels, in regulating epidermal hydration, barrier function, and cellular signalling. Among them, aquaporin-3 (AQP3), predominantly expressed in keratinocytes, has attracted particular attention due to its involvement in water and glycerol transport. Dysregulation of AQP expression has been associated with impaired barrier function, inflammatory skin disorders, and ageing. Growing evidence suggests that specific cosmetic ingredients and bioactive compounds, including glycerol, glyceryl glucoside, isosorbide dicaprylate, urea, retinoids, bakuchiol, peptides, plant extracts, and bacterial ferments, can modulate AQP3 expression, thereby improving skin hydration and resilience. Despite promising in vitro data, clinical evidence remains limited, mainly due to methodological and ethical constraints associated with assessing aquaporin expression in vivo. Nonetheless, aquaporins represent promising molecular targets for innovative cosmetic strategies aimed at enhancing hydration, promoting regeneration, and counteracting photoageing. Furthermore, AQP modulation may improve dermal delivery of active substances, providing new perspectives for advanced skincare formulation design. While the available evidence supports their cosmetic potential, emerging discussions on the safety of long-term AQP upregulation highlight the need for continued research and careful evaluation of such ingredients. Future studies should focus on elucidating the molecular mechanisms underlying AQP regulation and validating these findings in human clinical models. Full article

Type-P5 ATPases are the least characterized among the P-type ATPases and this is especially true in the case of the malaria parasite. In this study, Spf1, a subtype-P5A ATPase of yeast, and ATP13A2, a subtype-P5B ATPase of humans, were used as templates to extensively characterize the sequences and structural features of haemosporidian type-P5 ATPases. Malaria parasites have both subtype-P5A and subtype-P5B ATPase genes and the structural features of the proteins recapitulate the known structures of subtype-P5A and subtype-P5B ATPases. Detailed structural analysis detected an additional α-helix in the P-domain of subtype-P5A ATPases, which is not found in subtype-P5B ATPases. This feature may be an additional signature to distinguish subtype-P5A and subtype-P5B ATPases, in addition to the previously described differences in the membrane loops of the N-terminal domain, the arm in the P-domain of subtype-P5A, and substrate differences. A notable difference in the type-P5 ATPases from the malaria parasite, as compared to the templates, is the insertion of multiple variable and low-complexity regions that form intrinsically disorganized loops. These loops may form a shroud-like structure that protects the core ATPase structure and/or participates in low-affinity interprotein interactions. Homology modeling did not provide definitive answers about the substrate specificity of the haemosporidian type-P5 ATPases. However, the haemosporidian subtype-P5A ATPase is likely an ER transmembrane dislocase as are the other subtype-P5A ATPases. In contrast, the subtype-P5B ATPases of the malaria parasite are not likely to be polyamine transporters in lysosomes, as have been described in fungi and metazoans. This suggests that subtype-P5B ATPases have undergone lineage-specific divergence in regard to their function(s). Full article

Plant vacuolar-type Na⁺, K⁺/H⁺ antiporters (NHXs) play important roles in pH and K⁺ homeostasis and osmotic balance under normal physiological conditions. Under salt stress, vacuolar-type NHX enhances salt tolerance by compartmentalizing Na⁺ into vacuoles. However, the ion transport mechanism of vacuolar-type NHX remains poorly understood due to the absence of resolved protein crystal structures. To investigate the ion transport mechanism for vacuolar-type NHX, the three-dimensional structure of rice vacuolar-type NHX1 (OsNHX1) was established through homology modeling and AlphaFold3.0. The OsNHX1 model contains thirteen transmembrane segments according to hydrophobic characteristics and empirical and phylogenetic data. Furthermore, this study validated the OsNHX1 model via functional experiments, revealing a set of key charged amino acids essential for its activity. Mapping these amino acids onto the OsNHX1 model revealed that its pore domain exhibits a transmembrane charge-compensated pattern similar to that of NHE1 while also displaying a distinct charge distribution on either side of the pore domain. Comparative analysis of the key amino acid sites responsible for ion transport in the crystal structure of OsSOS1 and NHE1 revealed that OsNHX1 employs a unique ion transport mechanism. This study will enhance our understanding of the function and catalytic mechanism of OsNHX1 and other plant vacuolar-type NHXs. Full article

Modeling ion transport through membrane channels is crucial for understanding cellular processes, and Poisson–Nernst–Planck (PNP) equations provide a fundamental continuum framework for such ionic fluxes. We investigate a quasi-one-dimensional steady-state PNP system for two oppositely charged ion species, focusing on how large permanent charges within the channel and realistic boundary conditions impact ion transport. In contrast to classical models that impose ideal electroneutrality at the channel ends (a simplification that eliminates boundary layers near the membrane interfaces), we adopt relaxed neutral boundary conditions that allow small charge imbalances at the boundaries. Using asymptotic analysis treating the large permanent charge as a singular perturbation, we derive explicit first-order expansions for each ionic flux, incorporating boundary layer parameters $(\sigma ,\rho )$ to quantify slight deviations from electroneutrality. This analysis enables a qualitative characterization of individual cation and anion flux behaviors. Notably, we identify two critical transmembrane potentials, $V_{1c}$ and $V_{2c}$, at which the cation and anion fluxes, respectively, vanish, signifying flux-reversal thresholds that delineate distinct monotonic regimes in the flux-voltage response; these critical values depend on the permanent charge magnitude and the boundary layer parameters. We further show that both ionic fluxes exhibit saturation: as the applied voltage becomes extreme, each flux approaches a finite limiting value, with the saturation level modulated by the degree of boundary charge imbalance. Moreover, allowing even small boundary charge deviations reveals non-intuitive discrepancies in flux behavior relative to the ideal electroneutral case. For example, in certain parameter regimes, a large permanent charge that enhances an ionic current under strict electroneutral conditions will instead suppress that current under relaxed-neutral conditions (and vice versa). This new analytical framework exposes subtle yet essential nonlinear dynamics that classical electroneutral assumptions would otherwise obscure. It provides deeper insight into the interplay between large fixed charges and boundary-layer effects, emphasizing the importance of incorporating such realistic boundary conditions to ensure accurate modeling of ion transport through membrane channels. Numerical simulations are performed to provide more intuitive illustrations of our analytical results. Full article

Cystic fibrosis (CF), the most common hereditary lung disease in Caucasians, is caused by dysfunction of the cystic fibrosis transmembrane conductance regulator (CFTR). We evaluated CFTR function using a newly developed Ussing chamber system, the Multi Trans Epithelial Current Clamp (MTECC), in an in vitro model of human airway epithelia. Air–liquid interface (ALI) cultures were established from nasal brushings of healthy controls (HC) and CF patients with biallelic CFTR variants. ALI layer thickness was similar between groups (HC: 62 ± 13 µm; CF: 55 ± 9 µm). Immunofluorescence showed apical CFTR expression in HC, but reduced or absent signal in CF cultures. MTECC enabled continuous measurement of transepithelial resistance (Rt), potential difference (PD), and conductance (G_t). G_t was significantly reduced in CF cultures compared to HC (0.825 ± 0.024 vs. −0.054 ± 0.016 mS/cm²), indicating impaired cAMP-inducible ion transport by CFTR. Treatment of CF cultures with elexacaftor, tezacaftor, and ivacaftor (Trikafta^®) increased G_t, reflecting partial restoration of CFTR function. These findings demonstrate the utility of MTECC in detecting functional differences in CFTR activity and support its use as a platform for evaluating CFTR-modulating therapies. Our model may contribute to the development of personalized treatment strategies for CF patients. Full article

Bestrophinopathies are a group of inherited retinal diseases caused by mutations in the BEST1 gene. The protein encoded by this gene, bestorphin-1 (hBest1), is a calcium-dependent transmembrane channel localized on the basolateral membrane of retinal pigment epithelial (RPE) cells. We have already demonstrated the surface behavior and organization of recombinant hBest1 and its interactions with membrane lipids such as 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), sphingomyelin (SM) and cholesterol (Chol) in models of biological membranes, which affect the hBest1 structure–function relationship. The main aim of our current investigation is to integrate pure hBest1 protein into lipid bilayer nanostructures. We synthesized and characterized various hBest1-containing nanostructures based on 1,2-Dipalmitoylphosphatidylcholine (DPPC), SM, glycerol monooleate (GMO) and Chol in different ratios and determined their cytotoxicity and incorporation into cell membranes and/or cells by immunofluorescence staining. Our results show that these newly designed nanoparticles are not cytotoxic and that their incorporation into MDCK II cell membranes (used as a model system) may provide a mechanism that could be applied to RPE cells expressing mutated hBest1 in order to restore their ion transport functions, affected by mutated and malfunctioning hBest1 molecules. Full article

Zygotic genome activation (ZGA) marks the critical transition from reliance on maternal transcripts to the initiation of embryonic transcription early in development. Despite extensive characterization in model species, the regulatory framework of ZGA in sheep remains poorly defined. Here, we applied single-cell RNA sequencing (Smart-seq2) to in vivo- and in vitro-derived sheep embryos at the 8-, 16-, and 32-cell stages. Differential expression analysis revealed 114, 1628, and 1465 genes altered in the 8- vs. 16-, 16- vs. 32-, and 8- vs. 32-cell transitions, respectively, with the core pluripotency factors SOX2, NANOG, POU5F1, and KLF4 upregulated during major ZGA. To uncover coordinated regulatory modules, we constructed a weighted gene co-expression network using WGCNA, identifying the MEred module as most tightly correlated with developmental progression (r = 0.48, p = 8.6 × 10⁻¹⁴). The integration of MERed genes into the STRING v11 protein–protein interaction network furnished a high-confidence scaffold for community detection. Louvain partitioning delineated two discrete communities: Community 0 was enriched in ER–Golgi vesicle-mediated transport, transmembrane transport, and cytoskeletal dynamics, suggesting roles in membrane protein processing, secretion, and early signaling; Community 1 was enriched in G2/M cell-cycle transition and RNA splicing/processing, indicating a coordinated network for accurate post-ZGA cell division and transcript maturation. Together, these integrated analyses reveal a modular regulatory architecture underlying sheep ZGA and provide a framework for dissecting early embryonic development in this species. Full article

The interaction among the various Ca²⁺ transporters complicates the assessment of isolated systems in an intact cell. This article proposes the functionally isolated SR model (FISRM), a hybrid (experimental and mathematical) approach to study Ca²⁺ cycling between the cytosol and the sarcoplasmic reticulum (SR), the main source of Ca²⁺ for contraction in mammalian cardiomyocytes. In FISRM, the main transmembrane Ca²⁺ transport pathways are eliminated by using a Na⁺, Ca²⁺-free extracellular medium, and SR Ca²⁺ release is elicited by a train of brief caffeine pulses. Two compounds that exert opposite effects on the SR Ca²⁺ uptake were characterized by this approach in isolated rat ventricular cardiomyocytes. The experimental FISRM was simulated with a simple mathematical model of Ca²⁺ fluxes across the SR membrane, based on a previous model adapted to the present conditions. To a fair extent, the theoretical model could reproduce the experimental results, and confirm the main assumption of the experimental model: that the only relevant Ca²⁺ fluxes occur across the SR membrane. Thus, the FISRM seems to be a valuable framework to investigate the SR Ca²⁺ transport in intact cardiomyocytes under physiological and pathophysiological conditions, and to test therapeutic approaches targeting SR proteins. Full article

Edge-capping modified MXene membranes with new channels created by lateral nanosheets are of great research significance. After introducing tripolyphosphate (STPP) to Ti edges of Ti₃C₂T_x nanosheets and fabricating the STPP-MXene membranes edge-capping method, this research investigated the performance optimization mechanism of STPP-modified MXene membranes in terms of salt permeability (NaCl, Na₂SO₄, MgCl₂, and MgSO₄) and transmembrane energy barriers (E_salt) through the concentration gradient permeation test. Experimental results demonstrated an approximately 1.86-fold enhancement in salt flux (J_s) compared to the MXene membranes. The solution–diffusion model was also introduced to evaluate the salt solubility (K_s) and diffusivity (D_s) during permeation. Furthermore, analysis of transmembrane energy barriers revealed that STPP modification induced significantly larger reductions in activation energy for magnesium salts (MgSO₄: 55.1%; MgCl₂: 47.4%) compared to sodium salts (NaCl: 30.5%; Na₂SO₄: 30.9%). This phenomenon indicated the weakened electrostatic interactions between high-valent Mg²⁺ and the modified lateral membrane Ti edges, whereas the limited charge density of Na⁺ resulted in relatively modest optimization. The results highlight the contribution of STPP capping on the edges of adjacent lateral nanosheets. Therefore, the modification increased the transportation rate of cations across the MXene membrane by more than twice, thus advancing the application of 2D MXene membranes in resource recovery. Full article

There are two phenotypes in the natural populations of Marsupenaeus japonicus, which is an ideal model for studying the formation of markings and body color in crustaceans. In a previous study, we used comparative transcriptome technology to screen some functional genes related to body color regulation. Here, high-throughput sequencing technology was used to perform microRNA (miRNA) sequencing analysis on the exoskeleton of M. japonicus with two types of carapace markings, and functional studies of related genes were performed. A total of 687 mature miRNAs belonging to 135 miRNA families were identified in this study, and 111 novel miRNAs were found. Through stringent screening conditions, a total of 18 differentially expressed miRNAs were identified, including 14 with upregulated expression and 4 with downregulated expression. Multiple target genes were predicted for almost all of the differentially expressed miRNAs. The expression levels of several target genes, such as those related to cytoplasmic microtubule organization, transmembrane transportation, and signal transduction, were confirmed using qRT-PCR. This study revealed that both the CRCN A2 and CRCN C1 genes were highly expressed in type I individuals, while the expression levels of their related miRNAs in type I individuals were lower than those in type II individuals, which is consistent with the mechanism of miRNAs negatively regulating mRNA expression. Through interference with the CRCN A2 and CRCN C1 genes, a clear regulatory relationship was found between the two genes, and the dendritic xanthophores in the carapace of M. japonicus gradually changed from bright yellow to dark black, with obvious shrinkage. In summary, our studies provide references for the regulatory mechanisms of marking formation in M. japonicus. Full article