Search Results (125)

Background: The solute carrier family 22 member 23 (Slc22a23), a gene encoding a membrane-orphan transporter, is irreversibly upregulated in the brains of rats following fetal undernutrition. Materials and Method: To identify potential substrates of the SLC22A23 transporter, we analyzed the plasma of both Slc22a23-proficient and deficient rats. Results: We found that the level of lysophosphatidylcholine a C20:4 (LPC(20:4)) in plasma was more than twofold lower in the Slc22a23^−/− rats than in the Slc22a23^+/+ rats. We then conducted a series of animal behavioral tests on Slc22a23-proficient (Slc22a23^+/+) and deficient (Slc22a23^−/−) rats to investigate the effects of LPC(20:4) administration. We observed considerable effects in both Slc22a23^+/+ and Slc22a23^−/− rats, with more pronounced effects in the Slc22a23^+/+ rats. These effects included reductions in total distance travelled in an open field test, reductions in access to the novel objects in a novel object recognition test, reductions in sociability to a familiar rat in a social interaction test, and shorter latencies to the target in the Morris water maze test. Discussion: These findings suggest that LPC(20:4) administration enhances memory acquisition in both Slc22a23^+/+ and Slc22a23^−/− rats, though it appears to be more effective in the Slc22a23^+/+ rats. While the SLC22A23 transporter is likely involved in facilitating the transport of LPC(20:4), other transporters may also play a role in its transport in vivo. Full article

Malassezia are commensal lipid dependent yeasts which can cause opportunistic skin infection. Topical imidazole antifungals such as clotrimazole and ketoconazole are the frontline treatment. However, the tendency of fungal infections to recur, combined with the emergence of multi-azole-resistant Malassezia isolates means that many patients have used these antifungal treatments repeatedly or for extended durations with limited efficacy. While the impact of single azole treatments has been studied, the ability of specific azoles to induce cross-resistance is unclear. Understanding the effect of prior exposure of one treatment on susceptibility to other antifungals is important in the selection of the appropriate treatment to avoid driving the evolution of greater resistance. We previously identified drug transporters from the ATP-Binding Cassette (ABC) and Major Facilitator Superfamily (MFS) to be upregulated on extended exposure to clotrimazole. In this study, we investigated the effect of extended clotrimazole, ketoconazole and fluconazole exposure on antifungal cross-resistance profiles and examined the expression of the MFS transporters OPT1 and FLR1 in resistance emergence. We observed that treatment with clotrimazole was associated with increased cross-resistance to other antifungals. Ketoconazole treatment caused elevated MICs in all tested antifungals that did not decrease after drug removal. These findings advance our understanding of fungal adaptive resistance mechanisms and inform improved antifungal strategies to mitigate resistance development. Full article

Antimicrobial resistance (AMR) is a leading global cause of death, with recent World Health Organization (WHO) data revealing that one in six laboratory-confirmed bacterial infections shows resistance to at least one antibiotic treatment. This review comprehensively analyzes the AMR landscape in 2026, detailing its evolution, mechanisms, and the innovative strategies being deployed to combat it. Driven by Darwinian selection and accelerated by factors like antibiotic overuse during the Coronavirus Disease 2019 (COVID-19) pandemic (predominantly in hospitalized patients with suspected bacterial co-infection), AMR is propelled by a diverse molecular arsenal in bacteria. Key mechanisms include enzymatic drug inactivation (e.g., the diversifying β-lactamase superfamily), target site modification (e.g., mcr genes conferring colistin resistance), efflux pumps, and biofilm formation. The rapid global spread of these traits is facilitated by a dynamic “mobilome”, a network of plasmids and transposons that shuttle resistance genes between species. This crisis has sparked a major scientific mobilization. Advances include the discovery of novel antibiotic scaffolds like lariocidin and the regulatory approval of critical new antibiotic/inhibitor combinations such as sulbactam/durlobactam and aztreonam/avibactam, which target highly resistant Gram-negative bacteria. Moreover, the first-in-class antibiotic gepotidacin offers a new option for urinary tract infections. Beyond traditional drugs, the pipeline is diversifying to include phage therapy, antivirulence strategies, and artificial intelligence-guided drug discovery. This diversification is critical as it helps preserve the effectiveness of existing Medically Important Antimicrobials (MIAs), those deemed essential for human medicine, by providing alternative or adjunctive treatment options. However, scientific innovation alone is insufficient. This review argues that lasting success requires parallel progress in global policy and infrastructure. Strategic priorities beyond 2026 must include finalizing and funding updated global action plans, strengthening real-time surveillance and diagnostic capacity, especially in low-resource settings, and implementing new economic models to de-risk antibiotic development. Embedding effective antimicrobial stewardship within universal health coverage and pandemic preparedness plans is crucial. Ultimately, defeating AMR demands an unprecedented, coordinated global effort that outpaces the relentless adaptability of bacterial pathogens. Full article

Phytophthora infestans is the causal agent of late blight, one of the most destructive diseases of potato and tomato worldwide. Although qPCR-based methods are widely used to estimate pathogen biomass in infected tissues, methods for specifically assessing sporangial proliferation remain limited. In this study, we developed an RT-qPCR-based assay using PITG_13011, which encodes a predicted major facilitator superfamily transporter, as a sporangia-associated molecular marker in P. infestans. Among five candidate genes selected from transcriptomic data, PITG_13011 showed the strongest association with sporangia-associated samples in our validation assays. PITG_13011 transcripts were detectable from cDNA and genomic DNA derived from as few as 100 sporangia, and transcript abundance showed a strong positive correlation with sporangial number under controlled experimental conditions. In detached leaf inoculation assays, PITG_13011 transcript levels were associated with differences in sporangia-associated proliferation during infection. These results indicate that PITG_13011-based RT-qPCR can serve as a complementary molecular approach for estimating sporangia-associated proliferation of P. infestans in laboratory experiments. This method will be useful when sporangial production, rather than total pathogen biomass alone, is the parameter of interest. Full article

This study aimed to elucidate the molecular mechanisms underlying phenotypic divergence between two Lentinula edodes strains, Le_L and Le_S, which exhibit distinct fruiting body morphologies. While phenotypic variation among mushroom strains has been widely observed, the relative contributions of transcriptional regulation and structural genomic variation to these differences remain poorly understood. Comparative transcriptome analysis identified 8541 differentially expressed genes (DEGs), revealing clear functional divergence between the two strains. Genes upregulated in Le_S were predominantly enriched in ribosomal components and translation-related processes, indicating enhanced protein synthesis activity. In contrast, Le_L-upregulated genes were associated with transporters, transcription factors, and diverse metabolic pathways, suggesting broader regulatory and physiological functions. Protein–protein interaction network analysis further highlighted distinct regulatory architectures, with ribosomal proteins forming highly interconnected hub gene modules in Le_S, whereas Le_L hub genes were functionally diverse and included multiple members of the Major Facilitator Superfamily (MFS). Ortholog analysis across 33 L. edodes strains demonstrated that most hub genes were conserved, indicating their roles as core genetic components. Despite widespread genome-wide variation, including 7931 SNPs and 1149 INDELs, sequence variation within hub genes was limited, and allele-specific expression analysis revealed no significant allelic imbalance. In contrast, presence–absence variation (PAV) analysis identified structural differences affecting MFS transporter genes, which were absent in Le_S but present and upregulated in Le_L. Collectively, these findings suggest that structural genomic variation, particularly involving transporter genes, may play a more prominent role than sequence-level variation in driving phenotypic divergence. This study provides new insights into the genetic basis of strain-specific traits in L. edodes and highlights the importance of integrating multi-level genomic analyses. Full article

Yeast harbour more than ten different multiple drug resistance (MDR) genes encoding transporters that extrude xenobiotics from the cytoplasm into the environment. These transporters, belonging to the ATP-binding cassette (ABC) or major facilitator superfamily (MFS), exhibit broad and significantly overlapping substrate specificities, though the precise boundaries of their individual substrate ranges remain undefined. During evolution, genes with overlapping functions tend either to specialize or to degenerate into pseudogenes. Here, we propose several explanations for how this apparent redundancy of MDR efflux pumps benefits cells, and we discuss the potential individual roles of the full MDR efflux pump repertoire in the model organism Saccharomyces cerevisiae. We posit that individual MDR transporters may vary in stability under challenging environmental conditions, in the energetic cost of their synthesis and maintenance, and in their degree of specialization toward particular classes of xenobiotics. Furthermore, given that ABC transporters and MFS transporters exploit distinct driving forces for xenobiotic efflux, each class may have its own vulnerabilities. We argue that deciphering the distinct roles of MDR proteins will reveal critical weaknesses in the MDR system and guide the development of strategies to overcome multidrug resistance in pathogenic fungi. Full article

Although the ATP-binding cassette (ABC) transporter superfamily of proteins typically facilitates the movement of compounds across cellular membranes, the ABC E subfamily (ABCE) influences protein synthesis via non-transporter roles in ribosome biogenesis. Despite this essential role, our understanding of the impact that ABCE proteins have on insect physiology is limited. Here, we identified and characterized the ABCE gene from Lygus hesperus, a major agricultural pest of crops in North America. LhABCE transcripts were constitutively expressed throughout development and were present in all adult tissues tested. RNA interference (RNAi)-mediated knockdown of LhABCE transcripts in fifth instar nymphs resulted in high nymphal mortality and an incomplete molt. LhABCE knockdown in adults disrupted gametogenesis and reduced longevity. In females, oogenesis was impaired and oocytes did not progress beyond the pre-vitellogenic phase. In males, LhABCE knockdown reduced both spermatozoa abundance and male fertility. LhABCE knockdown, however, had little to no impact on hemolymph protein levels or the levels of circulating vitellogenin. Taken together, the results indicate that LhABCE is critical for the normal progression of processes like molting and gametogenesis that require coordinated bursts of protein synthesis and suggest that ABCE may play an important role in the mechanisms underlying those bursts. Full article

Hypocrellin A (HA) represents a pharmaceutically important perylenequinone photosensitizer produced by Shiraia bambusicola. However, submerged fermentation remains constrained by filamentous morphological characteristics and inherent mass transfer limitations. Although microparticle-enhanced cultivation (MPEC) has demonstrated efficacy in filamentous fungal systems, the molecular mechanisms by which physical cues, such as microparticle-induced shear stress, reprogram fungal metabolism remain largely unexplored. This work systematically optimizes SiO₂-based MPEC parameters for S. bambusicola GDMCC 60438, including particle dimensions, temporal addition protocols, and solid loading. Mechanistic investigations integrated pellet morphology analysis, membrane lipid composition, intracellular redox status, energy/precursor markers, and RNA-seq transcriptomic profiling with qRT-PCR validation. Under optimized conditions (10% w/v SiO₂, 30 mesh, added at 6 h), HA yield reached 41.76 ± 5.02 mg/L, representing a 3.65-fold increase over controls. MPEC shifted morphology toward smaller, more porous pellets with denser internal structure, accompanied by increased membrane fluidity (unsaturated/saturated fatty acid ratio from 1.54 to 2.63), elevated ROS levels with antioxidant enzyme activation, and enhanced acetyl-CoA and ATP accumulation. Transcriptomic analysis identified 206 differentially expressed genes enriched in oxidative phosphorylation, carbon metabolism, and stress responses, with upregulation of PKS-related biosynthetic genes and major facilitator superfamily transporters. This work establishes an integrated mechanistic framework linking particle-induced morphological changes to metabolic reprogramming through oxidative stress and subsequent transcriptional activation of the HA biosynthetic pathway, providing rational design principles for MPEC strategies in filamentous fungi. Full article

Boletus griseipurpureus is an ectomycorrhizal mushroom characterized by a bitter flavor. In this study, specimens were collected from three host plants—Acacia auriculiformis (BgAa), Melaleuca cajuputi (BgMc), and Eucalyptus camaldulensis (BgEc)—and initially classified based on pileus morphology. Molecular biodiversity was investigated using internal transcribed spacer (ITS) DNA barcoding, and comprehensive phylogenetic analysis revealed that B. griseipurpureus populations in southern Thailand clustered according to their symbiotic host species. De novo transcriptome assembly of B. griseipurpureus associated with different hosts was performed to generate unigene datasets, followed by functional gene annotation. A total of 1157 differentially expressed genes (DEGs) were identified and linked to ectomycorrhizal symbiosis. The genes involved in biosynthesis and metabolic processes exhibited host-dependent expression patterns. Furthermore, expression profiles of five selected genes—major facilitator superfamily (MFS) substrate transporter, phosphatase II, hexose transporter, terpenoid synthase, and fungal hydrophobin—were consistent between DEG analysis and semi-quantitative RT-PCR validation. These findings suggest that these genes play important roles in ectomycorrhizal symbiosis and the biosynthesis of bioactive compounds in B. griseipurpureus, with expression influenced by host association. This study provides valuable insights into the molecular biodiversity and gene regulation underlying ectomycorrhizal symbiosis, contributing to a better understanding of the biological processes in B. griseipurpureus. Full article

Cordycepin is a key active component of Cordyceps militaris, but the molecular mechanism underlying temperature-regulated biosynthesis remains unclear. In this study, Cordyceps militaris strain KN-1 was used as experimental material, with low-temperature (15 °C), control (20 °C), and high-temperature (25 °C) treatments applied during the fruiting body stage. Transcriptomics, untargeted metabolomics, weighted gene co-expression network analysis (WGCNA), and Reverse Transcription quantitative PCR (RT-qPCR) validation were integrated to elucidate the molecular mechanism of temperature-mediated cordycepin biosynthesis. The results showed that 25 °C increased fruiting body cordycepin content by 84%, while 15 °C reduced it. Transcriptomic analysis identified differentially expressed genes (DEGs) enriched in transmembrane transport and fatty acid metabolism, and untargeted metabolomics revealed differential metabolites (DAMs) enriched in lipids and organic acids, indicating that temperature primarily affects Cordyceps militaris membrane function. WGCNA showed that the MEblue module was positively correlated with cordycepin (r = 0.93), with Major Facilitator Superfamily (MFS) members accounting for the highest proportion (47.1%) that may affect cordycepin transmembrane transport. Multi-omics analysis indicated that high temperature promotes cordycepin accumulation through the synergistic regulation of multiple pathways: upregulating genes in the pentose phosphate pathway, purine metabolism, and cordycepin biosynthetic gene cluster (Cns1–Cns3), increasing protective agent pentostatin content, downregulating cordycepin-degrading genes, and enhancing cordycepin transmembrane transport. This study clarifies the molecular mechanism of temperature-mediated cordycepin accumulation, providing a theoretical basis for improving cordycepin production via temperature regulation, optimizing Cordyceps militaris strain quality, and facilitating efficient industrial production. Full article

Cell adhesion molecules (CAMs) are key regulators of tissue structural integrity and functional coordination, yet their specific role in the adaptation of yak lung tissue to high-altitude hypoxia remains unelucidated. Thus, we employed transcriptomic sequencing (RNA-seq), molecular biology assays, and single-cell RNA-seq (scRNA-seq) to analyze the expression characteristics of CAMs in yak lung tissues at high and low altitudes. Trypsin or collagenase digestion showed higher cell counts in high-altitude yak lungs (p < 0.05). RNA-seq analysis revealed significant enrichment of differentially expressed genes (DEGs) in adhesion-related pathways. Inductively coupled plasma mass spectrometry detected elevated Ca²⁺ levels in high-altitude yak lungs (p < 0.05). Quantitative real-time PCR (qRT-PCR) detection of key genes from five major families of CAMs revealed the downregulation of cadherin and integrin family-related genes, and upregulation of immunoglobulin superfamily-related genes, in high-altitude yak lungs (p < 0.05), corroborated by immunohistochemical (IHC) staining. A 10× scRNA-seq revealed adhesion changes in 9 of 15 lung cell subpopulations, with differentially expressed CAMs involving integrins. This study demonstrates that yak lung tissue establishes a sophisticated adhesive homeostasis through differential CAMs regulation. This strategy optimizes pulmonary immune responses and energy allocation, ensures structural integrity and functional coordination, and thereby facilitates superior acclimatization to higher-altitude hypoxia. Full article

Objective: This study compares the brain protective effects of L-borneolum and its main components (a combined application of L-borneol and L-camphor) on the rat model of middle cerebral artery occlusion/reperfusion (MCAO/R). It also makes clear the intrinsic regulatory mechanisms that link the neuroprotective effects of these compounds on IS to the blood-brain barrier (BBB), based on network pharmacology predictions. Furthermore, the study investigates the relationship between these compounds and the Major Facilitator Superfamily Domain-containing Protein 2A (MFSD2A)/Caveolin-1 (Cav-1) signaling axis. Methods: The MCAO/R model in rats was established to evaluate the therapeutic effect of L-borneolum (200 mg/kg) and its main components combination of L-borneol and L-camphor (6:4 ratio, 200 mg/kg). Neurological scores, 2,3,5-triphenyl tetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, and Nissl staining were performed to evaluate the neurological damage in the rats. Cerebral blood flow Doppler was applied to monitor the cerebral blood flow changes. Immunofluorescence analysis of albumin leakage and transmission electron microscopy (TEM) were conducted to evaluate blood-brain barrier (BBB) integrity. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the optimal drug concentration. Trans-epithelial electrical resistance (TEER) and horseradish peroxidase (HRP) assays were employed to confirm the successful establishment of an in vitro BBB co-culture model. Network pharmacology was utilized to predict the biological processes, molecular functions, and cellular components involved in the treatment of ischemic stroke (IS) by the main components of L-borneolum (L-borneol and L-camphor). Finally, immunofluorescence, real-time fluorescent quantitative PCR (RT-qPCR) and western blot analyses were performed to detect the expression of Major Facilitator Superfamily Domain Containing 2A (MFSD2A), caveolin-1 (CAV-1), sterol regulatory element-binding protein 1 (SREBP1) in brain tissue and hCMEC/D3 cells. Results: Network pharmacology prediction indicated that L-borneolum and its main components (L-borneol and L-camphor) in the treatment of IS are likely associated with vesicle transport and neuroprotection. Treatment of IS with L-borneolum and its main components significantly decreased neurological function scores and cerebral infarction area, while alleviating pathological morphological changes and increasing the number of Nissl bodies in the hippocampus. Additionally, it improved cerebral blood flow, reduced albumin leakage, and decreased vesicle counts in the brain. The trans-epithelial electrical resistance (TEER) of the co-culture model stabilized on the fifth day after co-culture, and the permeability to horseradish peroxidase (HRP) in the co-culture model was significantly lower than that of the blank chamber at this time. RT-qPCR and Western blot results demonstrated that, compared to the model group, the expression of SREBP1 and MFSD2A significantly increased, while the expression of Cav-1 decreased. Conclusions: L-borneolum and its main components combination (L-borneol/L-camphor, 6:4 ratio) may exert a protective effect in rats with IS by improving BBB transport function through modulation of the MFSD2A/Cav-1 signaling pathway. Full article

Sucrose is a key quality trait in peanuts, yet high-sucrose varieties are scarce. Although sucrose transporters (SUT/SUC) play crucial roles in sucrose transport and accumulation during seed development, systematic analyses in peanuts are limited. This study conducted a genome-wide analysis of the SUC gene family in cultivated peanut (Arachis hypogaea L.). Sixteen AhSUC genes were identified and characterized for genomic distribution, phylogeny, and expression across tissues and developmental stages. The genes are unevenly distributed across the genome with clustered chromosomal localization. All AhSUC proteins contain the conserved sucrose/proton co-transporter domain (IPR005989), exhibit the typical 12 transmembrane α-helical structure of the major facilitator superfamily, are hydrophobic, and predicted to localize to the membrane. Promoter analysis revealed cis-regulatory elements associated with growth, development, light, hormone, and stress responses. Expression profiling showed tissue-specific patterns, with eight AhSUC genes being highly expressed in cotyledons and embryos. Comparative analysis between high-sugar and conventional varieties showed higher expression of AhSUC2, AhSUC9, and AhSUC11 in the high-sugar variety, correlating with increased sucrose accumulation. Functional validation using a sucrose transport-deficient yeast mutant confirmed the sucrose transport activity of these genes. These findings provide insight into sucrose accumulation mechanisms and offer genetic targets for breeding high-sugar peanut varieties. Full article

Itaconic acid (IA), one of the top twelve renewable platform chemicals, is a key precursor for polymer synthesis. Here, we engineered Myceliophthora thermophila for efficient consolidated biocatalytic IA production from lignocellulose by introducing the heterologous IA pathway (cis-aconitic acid decarboxylase (CAD), mitochondrial tricarboxylic transporter (MTT), major facilitator superfamily transporter (MFS) from Aspergillus terreus), and boosting CAD expression and precursor supply. A critical issue was temperature mismatch: optimal fungal growth vs. CAD activity. Transcriptomics analysis identified reduced expression of glycolytic rate-limiting enzymes (fructose-bisphosphate aldolase, FBA; phosphofructokinase, PFK) at 40 °C. Overexpressing these enzymes in strain IA32 generated strain IA41 (with 3.1-fold and 2.8-fold higher expression of pfk and fba, respectively), which accelerated glucose consumption by 53.2% and increased IA yield by 55.1% A two-stage temperature-shift strategy (45 °C for growth/saccharification, 40 °C for CAD activity) was developed. The engineered strain achieved 3.93 g/L IA in shake flasks and 10.51 g/L in corncob fed-batch fermentation—the highest reported titer for consolidated lignocellulose-to-IA processes. This establishes M. thermophila as a robust platform for cost-effective IA production from lignocellulose. Full article

Detergent solubilisation remains the most commonly used but potentially problematic method to extract membrane proteins from lipid bilayers for Cryo-EM studies. Although recent advances have introduced excellent alternatives—such as amphipols, nanodiscs and SMALPs—the use of detergents is often necessary for intermediate steps. In this paper, we share our experiences working with detergent-solubilised samples within the modern Cryo-EM structural pipeline from the perspective of an EM specialist. Our aim is to inform novice users about potential challenges they may encounter. Drawing on specific examples from a variety of biological membrane systems, including Magnesium channels, lipopolysaccharide biosynthesis, and the human major facilitator superfamily transporters, we describe how the intrinsic properties of detergent-extracted samples can affect protein purification, Cryo-EM grid preparation (including the formation of vitreous ice) and the reconstitution of proteins into micelles. We also discuss how these unique characteristics can impact different stages of structural analysis and lead to complications in single-particle averaging software analysis. For each case, we present our insights into the underlying causes and suggest possible mitigations or alternative approaches. Full article