Search Results (5,183)

Cotton fiber quality improvement remains a fundamental challenge in breeding programs due to the complex genetic architecture underlying fiber development. The narrow genetic base of upland cotton (Gossypium hirsutum L.) and the quantitative nature of fiber quality traits necessitate innovative approaches for identifying and incorporating superior alleles from related species. We developed a BC₆F₂ population by introgressing chromosome segments from the sea island cotton variety Xinhai 36 (G. barbadense) into the upland cotton variety Xinluzhong 60 (G. hirsutum). Based on fiber strength phenotyping, we constructed two DNA bulks representing extreme phenotypes (20 superior and 12 inferior individuals) for bulked segregant analysis sequencing (BSA-Seq). High-throughput sequencing generated 225.13 Gb of raw data with average depths of 20× for parents and 30× for bulks. SNP calling and annotation were performed using GATK and ANNOVAR against the upland cotton reference genome (TM-1). BSA-Seq analysis identified 13 QTLs primarily clustered within a 1.6 Mb region (20.6–22.2 Mb) on chromosome A10. Within this region, we detected nonsynonymous mutation genes involving a total of six genes. GO and KEGG enrichment analyses revealed significant enrichment for carbohydrate metabolic processes, protein modification, and secondary metabolite biosynthesis pathways. Integration with transcriptome data prioritized GH_A10G1043, encoding a β-amylase family protein, as the key candidate gene. Functional validation through overexpression and RNAi knockdown in Arabidopsis thaliana demonstrated that GH_A10G1043 significantly regulates starch content and β-amylase activity, though without visible morphological alterations. This study successfully identified potential genomic regions and candidate genes associated with cotton fiber strength using chromosome segment substitution lines combined with BSA-Seq. The key candidate gene GH_A10G1043 provides a valuable target for marker-assisted selection in cotton breeding programs. Our findings establish a foundation for understanding the molecular mechanisms of fiber quality formation and offer genetic resources for developing superior cotton varieties with enhanced fiber strength. Full article

Skeletal muscle, the primary meat-producing tissue in bovines, is regulated by a complex transcriptional network during development. The role of Thrombospondin 3 (THBS3) and its associated super-enhancer (SE) in this process remains largely unknown. Here, by integrating multi-omics data, we identified THBS3 as a novel core regulator of myogenesis, orchestrated by a cognate super-enhancer (THBS3-SE). Functional assays demonstrated that THBS3 knockdown significantly promoted the proliferation and myogenic differentiation of bovine muscle stem cells (MuSCs) and accelerated their commitment to a fast-twitch fiber fate. Transcriptomic analysis linked THBS3 function to key signaling pathways controlling muscle growth, especially the mechanistic target of rapamycin (mTOR) signaling pathway. Mechanistically, we found that distal enhancers within the THBS3-SE loop to the THBS3 promoter drive its transcription, and CRISPR-based interference of these enhancers recapitulated the pro-myogenic effects of THBS3 knockdown. Collectively, our findings unveiled a THBS3-SE-mediated regulatory axis that critically governed bovine MuSCs’ fate. Targeting this axis may offer a novel strategy for improving beef production efficiency. Full article

Carboxylesterases (CCEs) have been demonstrated to be involved in pyrethroid resistance in insect species. This study aims to investigate CCE-mediated resistance mechanisms in Anopheles sinensis, a major malaria vector. Through comparative transcriptomics of a deltamethrin-resistant strain (CQ-LR) versus susceptible strain (WX-LS) of An. sinensis, we identified differentially expressed CCE genes across five developmental stages, five tissues, and three time points post-blood-meal. Four candidate genes (AsAe9, AsAe10, AsAce2, AsUn5) showed significantly upregulated expression. Subsequent qRT-PCR validation across four field-derived resistant strains (WX-LR, AH-LR, YH-LR, CQ-LR) and the susceptible strain confirmed significant upregulation of AsAe9, AsAe10, AsAce1, AsAce2, and AsBe4 in more than two resistant populations. RNAi-based functional validation showed that silencing AsAe10 or AsBe4 in the WX-LR strain significantly decreased knockdown time and raised 24 h mortality upon diagnostic deltamethrin exposure, with AsAe10 silencing having the strongest effect. This study identifies CCE genes involved in deltamethrin resistance in An. sinensis, providing valuable insights into the resistance mechanisms of pyrethroid and a theoretical basis for mosquito resistance management. Full article

Background and Objectives: Curcumin is a turmeric-derived polyphenol, and it has shown anticancer potential in various cancers, including breast cancer (BC). Nevertheless, the molecular mechanisms underlying its effects remain incompletely defined. Hsa_circ_0001946 (CDR1as) is a circular RNA (circRNA) that promotes tumor progression by competitively inhibiting microRNA-7-5p (miR-7-5p) in BC. This study investigated whether curcumin regulates the hsa_circ_0001946/miR-7-5p/target gene axis in BC progression. Materials and Methods: BC cell lines (MCF-7 and T47D) and a non-cancerous human mammary epithelial cell line (MCF-10A) were treated with curcumin or transfected with circ_0001946 siRNA or miR-7-5p mimic. Cell proliferation, migration, apoptosis, and protein expression were analyzed by CVDK-8 analysis, a wound healing assay, and flow cytometry, respectively. Also, protein expression levels were quantified via Western blotting. In vitro and in silico findings were further validated by analyzing tumor and adjacent normal tissues from 65 luminal BC patients. Results: Curcumin inhibited the proliferation and migration of MCF-7 and T47D cells in a dose-dependent manner. Knockdown of hsa_circ_0001946 or overexpression of miR-7-5p significantly suppressed proliferation and migration and enhanced apoptosis in BC cells compared to the negative controls. Curcumin treatment led to the knockdown of hsa_circ_0001946, the overexpression of miR-7-5p, and the downregulation of hsa_circ_0001946, CKS2, TOP2A, and PARP1, while it upregulating miR-7-5p. The Western blot confirmed reduced CKS2 protein levels after curcumin treatment. The expression of both hsa_circ_0001946 and CKS2 was significantly upregulated in tumor tissues compared to that of matched adjacent normal tissues, whereas that of miR-7-5p was markedly downregulated. Conclusions: This preliminary study shows that curcumin suppresses BC tumorigenesis by modulating the hsa_circ_0001946/miR-7-5p/target gene axis. While these findings suggest a novel regulatory pathway and potential therapeutic targets, further in vivo validation and clinical trials are required to determine the translational relevance of curcumin in BC therapy. Full article

Swiprosin-1 (SWS1/EFhd2) is a calcium-binding adaptor protein involved in cytoskeletal regulation, but its physiological role in bone homeostasis remains largely undefined. To elucidate its function in osteoclast biology, we examined SWS1 expression and activity during osteoclastogenesis using primary murine bone marrow-derived macrophages, siRNA-mediated knockdown, and SWS1 knockout (KO) mice. SWS1 was predominantly localized to the nucleus in precursor cells and redistributed to the F-actin ring in mature osteoclasts. Receptor activator of nuclear factor-kappa B ligand stimulation significantly downregulated SWS1 mRNA expression. Loss of SWS1 enhanced osteoclast formation, F-actin ring integrity, and bone resorption, accompanied by elevated expression of osteoclastogenic markers. In vivo, male SWS1 KO mice exhibited deteriorated trabecular bone microarchitecture with increased osteoclast numbers. Mechanistically, SWS1 deficiency intensified αvβ3 integrin-associated cytoskeletal signaling and upregulated Akt, MAPK, NF-κB, and PLCγ2 pathways. These results indicate that SWS1 negatively regulates osteoclast differentiation and function by restraining cytoskeletal reorganization and downstream signaling. Collectively, our findings establish SWS1 as a novel modulator of osteoclast activity and a potential therapeutic target for osteolytic bone disorders. Full article

Congenital ichthyoses, now grouped under the acronym EDD (Epidermal Differentiation Disorders), include nonsyndromic forms (nEDD) that may be caused by loss-of-function mutations in the CDSN gene encoding corneodesmosin (CDSN-nEDD, formerly Peeling skin syndrome type 1). It is characterized by skin peeling, inflammation, itching and food allergies, while no specific therapy is currently available. High levels of KLK5, the serine protease that initiates the desquamation cascade, are found in the epidermis of CDSN-nEDD patients. Thus, we hypothesized that KLK5 inhibition would alleviate the symptoms of CDSN-nEDD and could serve as a new pharmacological target. A human epidermal equivalent (HEE) model for CDSN-nEDD was developed using shRNA-mediated CDSN knockdown. This model was characterized and used to assess the role of KLK5 knockdown on CDSN-nEDD. Also, Klk5^−/− mice were crossed with Cdsn^epi−/− mice, the murine model of CDSN-nEDD, to examine in vivo the effect(s) of Klk5 deletion in CDSN-nEDD. Both models recapitulated the CDSN-nEDD desquamating phenotype. Elimination of KLK5 aggravated the CDSN-nEDD phenotype. Epidermal proteolysis was surprisingly elevated, while severe ultrastructural (corneo)desmosomal alterations increased epidermal barrier permeability and stratum corneum detachment was manifested. Based on these results, we concluded that targeting epidermal proteolysis with KLK5 ablation cannot compensate for the loss of corneodesmosin and rescue over-desquamation of the CDSN-nEDD. Possibly, in the absence of KLK5, other proteases take over which increases the severity of over-desquamation in CDSN-nEDD. The translational outcome is that over-desquamation may not always be rescued by eliminating epidermal proteolysis, but fine protease modulation is more likely required. Full article

The TREX-2 complex is conserved from yeast to humans and is responsible for mRNA export from the nucleus to the cytoplasm. In yeast and humans, the TPR and Nup153 nucleoporins of the nuclear pore complex are involved in TREX-2-dependent mRNA export, but data on their involvement in this process is rather controversial. In the present work, we have studied the role of TPR and Nup153 in the TREX-2-dependent export of hsp70 mRNA in Drosophila. We have shown that Nup153 and TPR are required for the TREX-2-dependent export of hsp70 mRNA, and their knockdown leads to mRNA accumulation in the cell nucleus. We have also demonstrated that Nup153 knockdown leads to TPR relocation to the nucleoplasm. Both nucleoporins are required for TREX-2 subunits’ association with the nuclear pore. Nup153 depletion leads to the TREX-2 subunits’ relocation from the nuclear pore to the nucleoplasm. The depletion of TPR leads to PCID2 relocation to the nucleoplasm and Xmas-2 retention at the nuclear pore and does not affect ENY2 redistribution. The TREX-2 subunits form several contacts with Nup153 and TPR. Hence, both nucleoporins are involved in the interaction with TREX-2 and TREX-2-dependent export in Drosophila. Full article

Metabolic dysregulation in the heart plays a critical role in the pathogenesis of atrial fibrillation (AF), yet the underlying molecular mechanisms remain unclear. Loss-of-function variants in the zinc finger homeobox 3 gene (ZFHX3) increase AF risk by promoting structural and electrical remodeling. However, the role of ZFHX3 knockdown (KD) in cardiac metabolism has not been fully elucidated. This study investigated the impact of ZFHX3 KD on energy metabolism in atrial myocytes and assessed the therapeutic potential of trimetazidine (TMZ). Seahorse XFe24 extracellular flux analysis, bioluminescent assays, microplate enzyme activity assays, and Western blotting were used to study energy substrate (glucose and fatty acid) oxidation stress, intracellular lactate content, glucose uptake, pyruvate dehydrogenase (PDH) activity, and regulatory protein expression in control and ZFHX3 KD HL-1 cells with or without TMZ (10 μM) treatment. ZFHX3 KD cells exhibited a higher acute response in oxygen consumption after Etomoxir injection, upregulated CD36 and phosphorylated ACC expression, increased glucose uptake and lactate production, reduced PDH activity, and higher levels of PDK4 and LDHA. Furthermore, ZFHX3 KD cells showed mitochondrial Ca²⁺ overload and increased phosphorylated PDH and oxidized CaMKII proteins, all of which were significantly attenuated by TMZ. Additionally, TMZ improved mitochondrial dysfunction in ZFHX3 KD cells by decreasing basal and maximal respiration, spare capacity, and proton leak. These findings suggest that ZFHX3 downregulation shifts substrate preference toward fatty acid utilization at the expense of glucose oxidation, contributing to metabolic and mitochondrial calcium dysregulation. TMZ mitigates these effects, highlighting its therapeutic potential in AF associated with ZFHX3 deficiency. Full article

Botulinum toxin type A (BTXA) is widely used for the treatment of sialorrhea; however, its mechanism remains unclear. Tight junctions (TJs) are limiting factors for salivary secretion through the paracellular pathway in the salivary gland, among which claudin-1 (Cldn1) is a TJ protein that mainly plays a barrier role. This study observed that Cldn1 was upregulated in BTXA-treated rats’ submandibular glands and SMG-C6 cells. Knockdown of Cldn1 reversed the BTXA-induced reduction in paracellular permeability. The transcription factor specificity protein-1 (Sp1), which binds to the Cldn1 promoter, was also upregulated by BTXA, and its expression was linked to the ERK1/2 pathway. Inhibition of ERK1/2 by U0126 reversed the BTXA-induced upregulation of Sp1 and Cldn1, as well as the reduction in paracellular permeability. MiR-124-3p, which directly targets Sp1, was downregulated by BTXA, but its overexpression counteracted Sp1 and Cldn1 upregulation. Although miR-124-3p did not affect ERK1/2 phosphorylation, ERK1/2 inhibition reversed the BTXA-induced decrease in miR-124-3p expression. These findings reveal a regulatory pathway through which BTXA reduces paracellular permeability in SMG-C6 cells via the ERK1/2/miR-124-3p/Sp1/Cldn1 axis. Full article

Midkine (MDK) is a multifunctional heparin-binding growth factor, and has been shown to regulate cell growth, survival, and migration. It also plays important roles in several inflammatory diseases such as sepsis. However, the role of MDK in the lungs has not yet been elucidated. In the present study, we investigated the role of MDK in pulmonary inflammation experiments using a mouse lipopolysaccharide (LPS)-induced pulmonary inflammation model and human bronchial cells. Wild-type and MDK-deficient mice were administered intratracheally with LPS, and several inflammatory parameters were analyzed. In the wild-type mice, MDK mRNA and protein in lung tissues were significantly increased after intratracheal LPS administration. The MDK-deficient mice showed significantly lower counts of total cells and neutrophils, as well as lower concentrations of total protein and neutrophil chemokines, KC and MIP-2 in bronchoalveolar lavage fluid, compared to wild-type mice. Moreover, mRNA expressions of TNF-α, keratinocyte chemoattractant (KC), and macrophage inflammatory protein (MIP)-2 in lung tissues, as well as the histopathological lung inflammation score, were significantly lower in the MDK-deficient mice. Furthermore, in in vitro experiments using bronchial epithelial cells, LPS stimulation increased mRNA expression of MDK, and MDK knockdown by siRNA decreased LPS-induced TNF-α and CXCL8 upregulation. These findings suggest that deficiency of MDK attenuates LPS-induced pulmonary inflammation, at least in part, through inhibiting inflammatory cytokine and chemokine upregulation in the lungs. Full article

Passion fruit (Passiflora edulis) is an economically important fruit worldwide. However, heat stress severely threatens its production, particularly in tropical and subtropical regions. To elucidate the molecular and metabolic mechanisms underlying heat tolerance, comparative physiological, transcriptomic, and metabolomic analyses were conducted between two yellow passion fruit cultivars: heat-tolerant ‘Summer Queen’ (F2) and heat-sensitive ‘Qinmi 9’ (QM9). Physiological evaluations demonstrated that QM9 exhibited significantly lower heat tolerance than F2, manifesting as severe leaf wilting, impaired photosynthetic efficiency, and elevated reactive oxygen species (ROS) accumulation. F2 exhibited distinct metabolic and transcriptional adaptations under heat stress, particularly in purine metabolism and flavonoid biosynthesis. Metabolites such as glutamine, xanthine, luteoloside, and trifolin were enriched in F2, alongside the upregulation of genes like adenosine kinase (AK), xanthine dehydrogenase (XDH), guanine deaminase (GDA), and flavonoid 3′-hydroxylase (F3′H). Weighted gene co-expression network analysis (WGCNA) highlighted strong associations between these pathways and transcription factors (e.g., MYB, HSF, WRKY), suggesting their pivotal roles in heat adaptation. Exogenous application of xanthine and trifolin markedly enhanced heat tolerance in passion fruit. Furthermore, knockdown of PeGDA and PeXDH markedly altered the heat tolerance of F2. These findings reveal that elevated metabolites in purine metabolism and flavonoid biosynthesis enhance heat tolerance in passion fruit, offering new insights into the molecular mechanisms of heat tolerance and potential targets for breeding climate-resilient passion fruit varieties. Full article

In Drosophila gonads, transposable elements (TEs) are repressed by the Piwi-interacting RNA (piRNA) pathway operating both co-transcriptionally and post-transcriptionally. In the non-gonadal tissues, TEs are mainly repressed by the short interfering RNA (siRNA) pathway with Argonaute 2 (Ago2) functioning as an effector protein. It is generally assumed that this pathway acts at the post-transcriptional level. However, recent data point to its possible involvement in co-transcriptional silencing as well. Here, using DamID, we found a drastic decrease in HP1a on TEs (especially on the LTR-containing retrotransposons) and other heterochromatin regions in Ago2-mutant Drosophila brain. HP1a reduction is accompanied by the increased chromatin accessibility of TEs, indicating their derepression. Accordingly, several LTR-containing retrotransposons were up-regulated in the larval brain of Ago2 mutants. Moreover, upon the knock-down of lamin Dm0 in neurons, HP1a was increased predominantly on the same set of TEs that had reduced HP1a binding in Ago2 mutants. We hypothesize that, since Ago2 was localized to the common complex with lamin Dm0, the depletion of the latter may release Ago2 in the nucleoplasm, thus enhancing the recruitment of HP1a on TEs. Our findings support the hypothesis that TEs in the Drosophila brain are silenced, in part, through Ago2-mediated recruitment of HP1a. Full article

Alterations in gene expression induced by ionizing radiation (IR) were commonly explained by transcriptional activation. However, the weak correlation between mRNA and protein levels following IR indicates the significant role for post-transcriptional regulation. microRNAs (miRNAs) bound to AGO2 play a significant role in post-transcriptional regulation; however, their role in radiation response is not clear. miRNA sequencing was performed to analyze the miRNAome of glioma cells. The effect of IR on Let-7 miRNAs and their association with AGO2 was examined using RT-qPCR and RNA immunoprecipitation (RIP) assays. Clonogenic assays were performed to measure radiosensitivity following Let-7a overexpression or knockdown. DNA damage (γH2AX foci) and cell cycle distribution were analyzed by immunofluorescence and flow cytometry. Let-7 miRNA regulatory networks were identified through target prediction and pathway enrichment analysis. AGO2-Let-7 binding decreased post IR, indicating impaired RISC loading. Let-7 overexpression increased radiosensitivity, DNA damage and G2/M cell cycle arrest in glioma and other cells (HeLa and MDA-MB-231). Let-7 miRNAs mainly targeted cell cycle and DNA damage response (DDR) pathways. Our study showed radiation impairs AGO2-miRNA binding, while restoring Let-7-AGO2 interaction enhances radiosensitivity by modulating DNA repair and cell cycle checkpoint activation. Targeting AGO2-miRNA dynamics represents a promising approach to improve radiotherapy outcomes. Full article

Purpose: We previously reported that the systemic administration of preprogrammed mouse hematopoietic bone marrow-derived progenitor cells (HSPCs) improved visual function and restored a functional retinal pigment epithelial (RPE) layer. Here, we investigated the potential impact of donor vs. host age on systemic cellular therapy in a murine model of retinal degeneration. Methods: HSPCs from young (8 weeks) and old (15 months) mice were programmed ex vivo with a lentiviral vector expressing the RPE65 gene (LV-RPE65) and systemically administering into young or old SOD2 KD mice. Visual loss and pathological changes were evaluated by electroretinogram (ERG), optical coherence tomography (OCT), histology, and immunohistochemistry. Results: Old donor HSPCs administered to old manganese superoxide dismutase (SOD2) knockdown (KD) recipient mice offered the least benefit. This was exemplified by the reduced recruitment and incorporation of LV-RPE65 HSPC into the RPE layer, as well as decreased improvement in visual function, retinal thinning, and limited reduction in oxidative damage and microglial activation. LV-RPE65 HSPC from young mice incorporated into the RPE layer of old SOD2 KD mice, though to a lesser extent than young cells administered to young hosts, offered some level of protection. By contrast, LV-RPE65 HSPCs from old mice, located to the subretinal space of young host mice, reduced visual loss, although some retinal pathology was observed. Conclusions: The administration of LV-RPE65 HSPC from old donors to old SOD2 KD mice offered the least improvement. Translational Relevance: Our findings highlight how both donor and recipient age impact the success of HSPC-based retinal therapy and using cells from aged donors for AMD treatment may have some limitations. Full article

Porcine Teschovirus (PTV) is a highly prevalent pathogen within swine populations, primarily associated with encephalitis, diarrhea, pneumonia, and reproductive disorders in pigs, thereby posing a significant threat to the sustainable development of the pig farming industry. In this study, a novel strain of PTV was isolated from the feces of a pig exhibiting symptoms of diarrhea, utilizing PK-15 cell lines. The structural integrity of the viral particles was confirmed via transmission electron microscopy, and the viral growth kinetics and characteristics were evaluated in PK-15 cells. High-throughput sequencing facilitated the acquisition of the complete viral genome, and subsequent phylogenetic analysis and full-genome alignment identified the strain as belonging to the PTV 2 genotype. Further investigation revealed that infection with the PTV-GXLZ2024 strain induces phosphorylation of the eukaryotic translation initiation factor 2α (eIF2α) in PK-15 cells, indicating activation of the unfolded protein response (UPR) through the PERK pathway, with minimal involvement of the IRE1 or ATF6 pathways. Notably, ATF4 protein expression was progressively downregulated throughout the infection, while downstream CHOP protein levels remained unchanged, indicating an incomplete UPR induced by PTV-GXLZ2024. Furthermore, PERK knockdown was found to enhance the replication of PTV-GXLZ2024. This study provides critical insights into the molecular mechanisms underlying PTV pathogenesis and establishes a foundation for future research into its evolutionary dynamics and interactions with host organisms. Full article