Search Results (58)

Leukodystrophies (LDs) constitute a heterogeneous group of genetic diseases in which mutations in glial cell genes lead to alterations in myelin formation and/or maintenance, ultimately causing white matter dysfunction. Increasing evidence on the genetic basis of LDs has revealed that proteins expressed not only by myelin-forming oligodendrocytes, but also by other glial cell types, play essential roles in myelination. By elucidating disease mechanisms, these studies have uncovered novel cellular and molecular contributors to myelin biogenesis and function, including ion channels. This is exemplified by the recent identification of mutations in the TMEM63A gene, which encodes the homonymous mechanosensitive channel, as the causative factor of the rare hypomyelinating LD HLD19 and by mutations in the chloride channel ClC-2 as responsible for the development of the vacuolating ClC2 LD. Together, this evidence has opened new perspectives on the crucial role of mechanosensitivity and ionic homeostasis for proper myelin development and structural integrity. In this review, we summarize recent advances on the role of glial ion channels in healthy white matter development and preservation, as well as their direct and indirect contributions to LD pathomechanisms. Finally, we discuss emerging therapeutic implications of these studies for LDs and other demyelinating conditions and emphasize the considerable potential of a cross-pathological, integrative approach to uncover shared and disease-specific mechanisms of demyelination. Full article

Variants in CLCN3 and CLCN4, encoding the neuronal endosomal Cl⁻/H⁺ antiporters ClC-3 and ClC-4, are linked to neurodevelopmental disorders with broad phenotypic variability. Over sixty CLCN4 variants have been functionally characterized, showing gain- or loss-of-function (GoF or LoF) effects. While ClC-3 can function as a homodimer, ClC-4 depends on heterodimerization with ClC-3 for efficient endosomal trafficking. CLCN4, located on the X chromosome, exhibits diverse pathogenic outcomes: complete LoF variants often cause non-syndromic presentations in hemizygous males and are asymptomatic in heterozygous females, whereas certain missense variants with partial or complete LoF produce severe syndromic phenotypes in both sexes. Here, we demonstrate dominant effects of three CLCN4 variants within ClC-3/ClC-4 heterodimers using two-electrode voltage-clamp recordings in Xenopus laevis oocytes and whole-cell patch-clamp recordings in mammalian cells co-expressing both proteins via a bicistronic IRES construct. Our findings provide the first evidence of dominant-negative CLCN4 effects within ClC-3/ClC-4 complexes and establish a platform for functional analysis of additional disease-associated variants. Full article

Soil salinization poses a significant threat to global agricultural productivity. Among crops, soybean (Glycine max), an important source of oil and protein, is more susceptible to salt stress compared to other major crops such as wheat (Triticum aestivum) and rice (Oryza sativa). To better utilize saline land resources, understanding the mechanisms underlying salt tolerance in soybean is essential for developing new salt-tolerant soybean varieties that contribute to food security. This review synthesizes current knowledge on the molecular mechanisms of salt tolerance in soybean, with a focus on ion homeostasis, osmotic adjustment, oxidative balance restoration, structural adaptations, and transcriptional regulatory networks. Key findings highlight the critical roles of ion transporters—such as GmNHX1, GmSOS1, GmHKT1, and GmCLC1—in maintaining Na⁺/K⁺ and Cl⁻ balance; the accumulation of osmoprotectants like proline and LEA proteins to alleviate osmotic stress; and the activation of antioxidant systems—including SOD, CAT, and APX—to scavenge reactive oxygen species (ROS). Additionally, structural adaptations, such as salt gland-like features observed in wild soybean (Glycine soja), and transcriptional regulation via ABA-dependent and independent pathways (e.g., GmDREB, GmbZIP132, GmNAC) further enhance tolerance. Despite these advances, critical gaps remain regarding Cl⁻ transport mechanisms, rhizosphere microbial interactions, and the genetic basis of natural variation in salt tolerance. Future research should integrate genomic tools, omics-based breeding, genome editing techniques such as CRISPR-Cas9, microbial technologies, and traditional breeding methods to develop salt-tolerant soybean varieties, providing sustainable solutions for the utilization of saline–alkali soils and enhancing global food security. Full article

Chloride channel proteins (CLCs) are essential anion transporters involved in plant growth, osmotic regulation, and ion homeostasis. However, their genome-wide characterization in tetraploid alfalfa (Medicago sativa L.) remains unexplored. In this study, a total of 35 CLC family members were identified and underwent comprehensive bioinformatic analyses. Phylogenetic and structural analyses divided them into six subfamilies and two subclasses based on conserved residues such as GxGIPE. Members within the same subclass shared conserved domains and similar motif patterns. Analysis of duplication events indicated that 48 segmental duplications were the primary driving force behind the expansion of this gene family. Promoter analysis revealed abundant light, hormone, and stress-responsive cis-elements, suggesting multiple regulatory functions. Gene expression profiling demonstrated that salt, drought stress, and ABA treatment significantly induced the expression levels of some genes. Among them, MsCLC2 and MsCLC18 from Group c exhibited more than fivefold upregulation under both salt and drought stress, significantly higher than other members. Subcellular localization confirmed MsCLC18 on the plasma membrane, potentially regulating Cl⁻ efflux through a Cl⁻/H⁺ antiporter mechanism to alleviate Cl⁻ toxicity. These findings provide a theoretical foundation for the function study of CLC genes in alfalfa and offer new insights into the molecular evolution of polyploid plants under abiotic stress. Full article

Background/Objectives: Patients with liver cirrhosis (LC) present hematological abnormalities with multiple etiologies. As liver function deteriorates, these abnormalities become more evident, conferring a higher risk of morbidity and mortality. The objective of this study was to determine the hematological alterations present in patients with compensated vs. decompensated liver cirrhosis. Methods: This study is a prospective study conducted in a tertiary hospital with patients diagnosed with compensated liver cirrhosis monitored by the Gastroenterology Department, in addition to patients hospitalized for decompensated liver cirrhosis. Serum samples were taken after an informed consent form was signed, and clinical and biochemical data were recorded. Results: Seventy-three percent of patients with decompensated liver cirrhosis (DLC) suffered from anemia and thrombocytopenia at the start of the study. Patients with LC are at greater risk of developing leukopenia/lymphopenia if they are in a more advanced stage of the disease, and the erythrocyte sedimentation rate (ESR) and C-reactive protein levels are higher in this group than in patients with compensated LC. Twenty-eight percent (4/14) of patients with DLC were admitted due to evidence of acute gastrointestinal bleeding; the remaining 18 patients suffering from CLC and DLC were recorded as having an average hemoglobin level of 11 g/dL with no evidence of bleeding. Conclusions: Hematological abnormalities are common in patients with LC, particularly in the advanced stages of the disease, and these abnormalities increase the risk of morbidity and mortality. The authors have read the CONSORT 2010 Statement, and the manuscript was prepared and revised in accordance. Full article

Psoriasis is a complex inflammatory disease related to cardiometabolic disorders (CMDs). Galectin-4 (gal-4) is involved in biological processes such as lipid raft stabilization, intestinal inflammation and tumorigenesis. Charcot-Leyden crystals (CLCs), inter alia, Charcot-Leyden crystal/galectin-10 (CLC/gal-10), are involved in eosinophil-derived diseases. To date, neither of these galectins has been investigated in the context of psoriasis. The study aimed to evaluate serum galectin-4 and -10 levels in psoriatic patients and explore potential relationships with disease activity, metabolic or inflammatory indices. Blood samples were collected from 60 patients with plaque-type psoriasis and 30 healthy volunteers and evaluated using an Enzyme-Linked Immunosorbent Assay (ELISA). Morphological and biochemical indices were measured using routine laboratory techniques. Plasma gal-4 and gal-10 concentrations were significantly higher in patients with psoriasis than in the control group (p < 0.05). Galectins did not correlate with the Psoriasis Area Severity Index (PASI) nor age (p > 0.5); however, gal-4 showed a significant positive correlation with Body Mass Index (BMI), psoriasis duration (p = 0.03), and transaminase activity. Both proteins were the highest in obese psoriatics (p < 0.05). The results indicate that galectin-4 and galectin-10 may be involved in the pathophysiological mechanisms underlying CMDs in psoriatics. These galectins represent promising candidates for biomarkers of metabolically driven inflammation, with galectin-4, in particular, emerging as a potential indicator of hepatic dysfunction in psoriatic patients. Full article

Lymphoma is the most common neoplasm of lymphoid tissues in dogs. Exportin 1 (XPO1) is an important major nuclear receptor for exporting proteins and RNA species. The XPO1 upregulation can eliminate some tumor suppressor proteins (TSPs) function upon their nuclear–cytoplasmic export. The XPO1 inhibitor, KPT-335, blocks the translocation of TSPs and restores their function to induce cell cycle arrest, apoptosis, and cell proliferation. This in vitro study aimed to evaluate the XPO1 mRNA and protein expression in canine lymphoma cell lines and confirm the relevance with KPT-335. XPO1 mRNA and protein levels were quantified, and the effect of KPT-335 was assessed by a cell proliferation assay. The results indicated that XPO1 mRNA and protein were highly expressed in 17-71, CLBL-1, CLC, CLGL-90, and UL-1, and were moderately expressed in GL-1, Ema, and Nody-1. All canine lymphoma cell lines showed dose-dependent growth inhibition and decreased cell viability in response to KPT-335, with IC₅₀ concentrations ranged from 89.8–418 nM. The expression levels of XPO1 mRNA and protein were related; however, no correlation was found between those expression levels and the efficacy of KPT-335. These findings suggest that XPO1 may represent a promising target for therapeutic intervention in canine lymphoma. Full article

Oxidative stress is a key mediator of physiological dysfunction in aquatic organisms under environmental challenges, yet its comprehensive impacts on gill physiology require further clarification. This study investigated the molecular and cellular responses of Eriocheir sinensis gills to hydrogen peroxide (H₂O₂)-induced oxidative stress, integrating antioxidant defense, ion transport regulation, and stress-induced cell apoptosis and autophagy. Morphological alterations in the gill filaments were observed, characterized by septum degeneration, accumulation of haemolymph cells, and pronounced swelling. For antioxidant enzymes like catalase (CAT) and glutathione peroxidase (GPx), activities were enhanced, while superoxide dismutase (SOD) activity was reduced following 48 h of exposure. Overall, the total antioxidant capacity (T-AOC) showed a significant increase. The elevated concentrations of malondialdehyde (MDA) and H₂O₂ indicated oxidative stress. Ion transport genes displayed distinct transcription patterns: Na⁺-K⁺-2Cl⁻ co-transporter-1 (NKCC1), Na⁺/H⁺ exchanger 3 (NHE3), aquaporin 7 (AQP7), and chloride channel protein 2 (CLC2) were significantly upregulated; the α-subunit of Na⁺/K⁺-ATPase (NKAα) and carbonic anhydrase (CA) displayed an initial increase followed by decline; whereas vacuolar-type ATPase (VATP) consistently decreased, suggesting compensatory mechanisms to maintain osmotic balance. Concurrently, H₂O₂ triggered apoptosis (Bcl2, Caspase-3/8) and autophagy (beclin-1, ATG7), likely mediated by MAPK and AMPK signaling pathways. These findings reveal a coordinated yet adaptive response of crab gills to oxidative stress, providing new insights into the mechanistic basis of environmental stress tolerance in crustaceans. Full article

Background: The pathogenesis of neuropsychiatric lupus erythematosus (NPSLE) is very complex and is associated with neuroinflammation and blood–brain barrier compromise. Experimental investigations of NPSLE have classically relied on spontaneous models. Recently, TLR7 agonist-induced lupus has been shown to exhibit similar neuropsychiatric manifestations to spontaneous ones. Cinnamon is a widespread spice and natural flavoring agent. It has been proven to modulate vascular endothelial tight junctions, neuroinflammation, and autoimmunity pathways, but it has never been tested in relation to lupus. Hypothesis/Purpose: In this pilot study, we aimed to explore the disease-modifying effect of Cinnamomum cassia on NPSLE in a TLR7 agonist-induced model. Study Design: An experimental design was followed in this study. Methods: Lupus was induced in C57BL/6J female mice via the direct application of imiquimod, a TLR7 agonist (5% imiquimod cream, 1.25 mg three times weekly), to the skin. Mice were divided into five groups (n = 8 per group): a sham group (S), a sham group supplemented with cinnamon (SC), an imiquimod-treated group (L), an imiquimod-treated group supplemented with cinnamon starting from induction (LC), and an imiquimod-treated group supplemented with cinnamon beginning two weeks prior to induction (CLC). This protocol was followed for six consecutive weeks. Cinnamomum cassia powder was administered orally at 200 mg/kg, 5 days per week. Results: Behavioral alterations were significantly ameliorated in the CLC group compared to lupus mice. Neuronal shrinkage and nuclear chromatin condensation were visible in the hippocampal cornu ammonis and dentate gyrus zones of lupus mice, with an increased expression of TLR7 and NLRP3, versus significantly less neurodegeneration and TLR7 and NLRP3 expression in the CLC group. In addition, the expression of the blood–brain barrier endothelial cell tight junction proteins claudin-1, occludin, and ZO-1 was abnormally modified in lupus mice and was restored in the CLC group. Moreover, while the cell–cell border delocalization of claudin-1 was documented in cultured blood–brain barrier endothelial cells treated with the plasma of lupus mice to a punctate intracytoplasmic fluorescence pattern, only cells treated with the plasma of the CLC group exhibited a complete reversal of this redistribution of claudin-1. Finally, cinnamaldehyde seemed to interact with TLR7 at multiple sites. Conclusions:Cinnamomum cassia seems to alleviate the pathogenesis of NPSLE. Supplementation with Cinnamomum cassia could be of great interest to modulate the activity and severity of the disease. Full article

Artemisia dracunculus/(tarragon) is a perennial plant used in traditional medicine and the food industry. The plant is known to have beneficial effects on health, such as antibacterial, antifungal, antiseptic, carminative, anti-inflammatory, antipyretic, anthelmintic, etc. In this study, the compounds present in the highest concentrations in the essential oils obtained by different extraction methods from tarragon found on the Romanian market were identified by gas chromatography–mass spectrometry. The biological activity of these compounds was predicted using the computational tools ADMETlab3.0, admetSAR3.0, CLC-Pred2.0, and AntiBac-Pred. Also, the main molecular target of these compounds was predicted and the interactions with this protein were evaluated using molecular docking. The compounds identified in high concentrations in the obtained essential oils are estragole, cis-β-ocimene, trans-β-ocimene, limonene, eugenol methyl ether, eugenol acetate, eugenol, caryophyllene oxide, and α-pinene. The absorption, distribution, metabolism, excretion, and toxicity profiles of these compounds show that they are generally safe, but some of them can cause skin sensitization and respiratory toxicity and are potential inhibitors of the organic anion transporters OATP1 and OATP2. Several of these compounds exert antibacterial activity against some species of Staphylococcus, Streptococcus, and Prevotella. All compounds reveal potential cytotoxicity for several types of cancer cells. These findings may guide further experimental studies to identify medical and pharmacological applications of tarragon extracts or specific compounds that can be isolated from these extracts. Full article

Background: Chemoresistance in triple-negative breast cancer (TNBC) presents a significant clinical hurdle, limiting the efficacy of treatments like doxorubicin. This study aimed to explore the molecular changes associated with doxorubicin resistance and identify potential therapeutic targets to overcome this resistance, thereby improving treatment outcomes for TNBC patients. Methods: Doxorubicin-resistant (DoxR) TNBC models (MDA-MB-231 and BT-549) were generated by exposing cells to increasing concentrations of doxorubicin. RNA sequencing (RNA-Seq) was performed using the Illumina platform, followed by bioinformatics analysis with CLC Genomics Workbench and iDEP. Functional assays assessed proliferation, sphere formation, migration, and cell cycle changes. Protein expression and phosphorylation were confirmed via Western blotting. Pathway and network analyses were conducted using Ingenuity Pathway Analysis (IPA) and STRING, while survival analysis was performed using Kaplan–Meier Plotter database. Results: DoxR cells exhibited reduced proliferation, sphere formation, and migration, but showed enhanced tolerance to doxorubicin. Increased CHK2 and p53 phosphorylation indicated cellular dormancy as a resistance mechanism. RNA-Seq analysis revealed upregulation of cytokine signaling and stress-response pathways, while cholesterol and lipid biosynthesis were suppressed. Activation of the IL1β cytokine network was prominent in DoxR cells, and CRISPR-Cas9 screens data identified dependencies on genes involved in rRNA biogenesis and metabolism. A 27-gene signature associated with doxorubicin resistance was linked to worse clinical outcomes in a large breast cancer cohort (HR = 1.76, FDR p < 2.0 × 10⁻¹³). Conclusions: This study uncovers potential therapeutic strategies for overcoming TNBC resistance, including dormancy reversal and targeting onco-ribosomal pathways and cytokine signaling networks, to improve the efficacy of doxorubicin-based treatments. Full article

Protists inhabit marine, brackish and fresh waters. The salt barrier plays an important role in the origin of their diversity. Salinity tolerance differs among species and sometimes even among different strains of the same species, indicating local adaptation. Dinoflagellates from the Apocalathium genus are represented by at least four species, which originated via rapid and recent radiation. Water salinity was suggested as one of the key factors for this radiation. A previous study found RNA transcripts, which belong exclusively to saline strains of Apocalathium, and were absent in its freshwater strains. In the present paper, the diversity of these transcripts and their orthologs from marine and freshwater protists were analysed using bioinformatic approaches. First, it was found that these specific transcripts translated to the proteins, which are important for osmoregulation (e.g., transport of various compounds including glycine betaine, regulation of microtubule organisation, post transcriptional modifications). This supports the idea that speciation within Apocalathium resulted in the loss of osmoregulatory genes by freshwater species. Second, protein distribution was not highly species specific, because their orthologs were found in different dinoflagellates and were relatively common in other phototrophic protists, though the sequences were highly variable. Proteins from 13 orthogroups were absent or very rare in studied freshwater genomes and transcriptomes. They could play a specific role in protists salinity tolerance. Third, detailed phylogenetic analyses of betaine-like transporter and chloride transmembrane transporters, which probably are one of the key proteins associated with salinity tolerance, revealed high levels of multiple and variable copies that were not eliminated from the genome during the evolution. The expression of their genes could be important in the adaptation of dinoflagellates to salinity changes, as it was already shown for some other protists. Full article

Dent disease type 1 is a rare X-linked recessive inherited renal disorder affecting mainly young males, generally leading to end-stage renal failure and for which there is no cure. It is caused by inactivating mutations in the gene encoding ClC-5, a 2Cl⁻/H⁺ exchanger found on endosomes in the renal proximal tubule. This transporter participates in reabsorbing all filtered plasma proteins, which justifies why proteinuria is commonly observed when ClC-5 is defective. In the context of Dent disease type 1, a proximal tubule dedifferentiation was shown to be accompanied by a dysfunctional cell metabolism. However, the exact mechanisms linking such alterations to chronic kidney disease are still unclear. In this review, we gather knowledge from several Dent disease type 1 models to summarize the current hypotheses generated to understand the progression of this disorder. We also highlight some urinary biomarkers for Dent disease type 1 suggested in different studies. Full article

Chloride channels (ClCs) have received global interest due to their significant role in the regulation of ion homeostasis, fluid transport, and electrical excitability of tissues and organs in different mammals and contributing to various functions, such as neuronal signaling, muscle contraction, and regulating the electrolytes’ balance in kidneys and other organs. In order to define the chloride voltage-gated channel (CLCN) gene family in buffalo, this study used in silico analyses to examine physicochemical properties, evolutionary patterns, and genome-wide identification. We identified eight CLCN genes in buffalo. The ProtParam tool analysis identified a number of important physicochemical properties of these proteins, including hydrophilicity, thermostability, in vitro instability, and basic nature. Based on their evolutionary relationships, a phylogenetic analysis divided the eight discovered genes into three subfamilies. Furthermore, a gene structure analysis, motif patterns, and conserved domains using TBtool demonstrated the significant conservation of this gene family among selected species over the course of evolution. A comparative amino acid analysis using ClustalW revealed similarities and differences between buffalo and cattle CLCN proteins. Three duplicated gene pairs were identified, all of which were segmental duplications except for CLCN4-CLCN5, which was a tandem duplication in buffalo. For each gene pair, the Ka/Ks test ratio findings showed that none of the ratios was more than one, indicating that these proteins were likely subject to positive selection. A synteny analysis confirmed a conserved pattern of genomic blocks between buffalo and cattle. Transcriptional control in cells relies on the binding of transcription factors to specific sites in the genome. The number of transcription factor binding sites (TFBSs) was higher in cattle compared to buffalo. Five main recombination breakpoints were identified at various places in the recombination analysis. The outcomes of our study provide new knowledge about the CLCN gene family in buffalo and open the door for further research on candidate genes in vertebrates through genome-wide studies. Full article

The nine-member CLC gene family of Cl⁻ chloride-transporting membrane proteins is divided into plasma membrane-localized Cl⁻ channels and endo-/lysosomal Cl⁻/H⁺ antiporters. Accessory proteins have been identified for ClC-K and ClC-2 channels and for the lysosomal ClC-7, but not the other CLCs. Here, we identified TMEM9 Domain Family Member B (TMEM9B), a single-span type I transmembrane protein of unknown function, to strongly interact with the neuronal endosomal ClC-3 and ClC-4 transporters. Co-expression of TMEM9B with ClC-3 or ClC-4 dramatically reduced transporter activity in Xenopus oocytes and transfected HEK cells. For ClC-3, TMEM9B also induced a slow component in the kinetics of the activation time course, suggesting direct interaction. Currents mediated by ClC-7 were hardly affected by TMEM9B, and ClC-1 currents were only slightly reduced, demonstrating specific interaction with ClC-3 and ClC-4. We obtained strong evidence for direct interaction by detecting significant Förster Resonance Energy Transfer (FRET), exploiting fluorescence lifetime microscopy-based (FLIM-FRET) techniques between TMEM9B and ClC-3 and ClC-4, but hardly any FRET with ClC-1 or ClC-7. The discovery of TMEM9B as a novel interaction partner of ClC-3 and ClC-4 might have important implications for the physiological role of these transporters in neuronal endosomal homeostasis and for a better understanding of the pathological mechanisms in CLCN3- and CLCN4-related pathological conditions. Full article