ATP-binding Cassette Research Articles

Crosslink among cyclin-dependent kinase 9, ATP binding cassette transporter G2 and Beclin 1 in colorectal cancer

Colorectal cancer (CRC) ranks third in the number of cancers mainly because of the inability to diagnose it at an early stage. The pathogenesis of CRC is complicated, which is the result of the complex interaction of multiple genetic and environmental factors. Currently, one of the main treatments for CRC is chemotherapy. But the primary cause of CRC treatment failure is drug resistance. The expression of cyclin-dependent kinase 9 (CDK9) was correlated with elevated autophagy levels in colon cancer, and high expression of CDK9 indicates a poor prognosis in CRC. The incidence of autophagy and the expressions of Beclin 1 and ATP binding cassette transporter G2 are different in left and right colon cancer, and autophagy may be involved in the occurrence of chemotherapy resistance. In this article, the roles of CDK9, ATP binding cassette transporter G2 and Beclin 1 in CRC were elucidated, emphasizing the linkages among them and providing potential therapeutic targets of CRC.

Genomic Characterizations of Klebsiella variicola: Emerging Pathogens Identified from Sepsis Patients in Ethiopian Referral Hospitals

Healthcare in low- and middle-income countries is becoming problematic due to the emergence of multidrug-resistant bacteria causing serious morbidity and mortality. Klebsiella variicola carrying multiple antimicrobial resistance (AMR) genes was found significantly among sepsis patients in a study done between October 2019 and September 2020 at four Ethiopian hospitals located in the central (Tikur Anbessa and Yekatit 12), southern (Hawassa), and northern (Dessie) parts. Among 1416 sepsis patients, 74 K. variicola isolates were identified using MALDI-TOF, most of them at Dessie (n=44) and Hawassa (n=28) hospitals. Whole genome sequencing showed that K. variicola strains identified at Dessie Hospital displayed phylogenetic clonality, carried an IncM1 plasmid and the majority were ST3924. Many K. variicola identified at Hawassa Hospital were clonally clustered and the majority belonged to novel STs and carried IncFIB(K) and IncFII(K) plasmids concurrently. Fifty K. variicola carried ESBL genes while 2 isolates harbored AmpC. Other frequently found genes were aac(3)-lla, bla CTX-M-15, bla TEM-1B, bla LEN2, bla OXA-1, bla SCO-1, catB3, dfrA14, QnrB1, aac(6')-lb-cr and sul2. Virulence genes detected at both sites were mrk operons for biofilm formation and siderophore ABC transporter operons for iron uptake. Capsular alleles varied, with wzi 269 at Dessie and wzi 582 at Hawassa. The isolation of multidrug-resistant K. variicola as an emerging sepsis pathogen calls for strong infection prevention strategies and antimicrobial stewardship supported by advanced bacterial identification techniques.

Stable Isotope and Multiomics Reveal Uptake, Translocation, and Transformation Mechanisms of Tris(2-chloroethyl) Phosphate in Wheat (Triticum aestivum L.).

Uptake, translocation, and transformation mechanisms of tris(2-chloroethyl) phosphate (TCEP) in hydroponic wheat (Triticum aestivum L.) were systematically investigated using compound-specific stable isotope and multiomics analyses in this study. Results showed that TCEP was quickly adsorbed on root epidermis and then absorbed in roots via water and anion channels as well as an active process dependent on energy. Active process and anion channel preferentially translocated TCEP-containing light carbon isotopes and dominated the transmembrane transport of TCEP to enter vascular bundle. Transcriptomic and metabolomic analyses indicated gene-encoding ATP-binding cassette (ABC) transporters and purple acid phosphatases (PAPs) and glutathione S-transferases (GSTs) involved in TCEP transport and transformation, respectively. Molecular docking simulations showed that TCEP bound to the hydrophilic cavity of ABC transporter/PAP and hydrophobic cavity of GST, and hydrogen bonding was the important driving force. The results of this study offered insights for future effective mitigation of TCEP risk in edible plants.

Transcriptome-guided breeding for Paspalum notatum: producing apomictic hybrids with enhanced omega-3 content

Key messageTranscriptomics- and FAME-GC-MS-assisted apomixis breeding generated Paspalum notatum hybrids with clonal reproduction and increased α-linolenic acid content, offering the potential to enhance livestock product's nutritional quality and reduce methane emissions A low omega-6:omega-3 fatty acid ratio is considered an indicator of the nutritional impact of milk fat on human health. In ruminants, major long-chain fatty acids, such as linoleic acid (18:2, omega-6) and α-linolenic acid (18:3, omega-3), originate from dietary sources and reach the milk via the bloodstream. Since forages are the primary source of long-chain fatty acids for such animals, they are potential targets for improving milk lipid composition. Moreover, a high 18:3 content in their diet is associated with reduced methane emissions during grazing. This work aimed to develop genotypes of the forage grass Paspalum notatum with high leaf 18:3 content and the ability for clonal reproduction via seeds (apomixis). We assembled diploid and polyploid Paspalum notatum leaf transcriptomes and recovered sequences of two metabolism genes associated with the establishment of lipid profiles, namely SUGAR-DEPENDENT 1 (SDP1) and PEROXISOMAL ABC TRANSPORTER 1 (PXA1). Primers were designed to amplify all expressed paralogs in leaves. qPCR was used to analyse SDP1 and PXA1 expression in seven divergent genotypes. Reduced levels of SDP1 and PXA1 were found in the polyploid sexual genotype Q4188. Fatty acid methyl esters/gas chromatography/mass spectrometry (FAME/GC/MS) assays confirmed an increased percentage of 18:3 in this genotype. Crosses between Q4188 and the obligate apomictic pollen donor Q4117 resulted in two apomictic F1 hybrids (JS9 and JS71) with reduced SDP1 and PXA1 levels, increased 18:3 content, and clonal maternal reproduction. These materials could enhance milk and meat quality while reducing greenhouse gas emissions during grazing.

LysR-Type Transcriptional Regulator Contributes to Pseudomonas cannabina pv. alisalensis Virulence by Regulating Type Three Secretion System

Pseudomonas cannabina pv. alisalensis (Pcal) causes bacterial blight on cabbage. In a previous study, we screened for reduced virulence using Tn5 transposon mutants and identified a LysR-type transcriptional regulator (LTTR) as a potential virulence factor in Pcal. However, the role of LTTR in Pcal virulence has not been thoroughly investigated. In this study, we demonstrated that the Pcal NN14 mutant (with Tn5 insertion in the LTTR-encoding gene) showed reduced disease symptoms and bacterial populations in cabbage, indicating that LTTR contributes to Pcal virulence. RNA-seq analysis identified 39 LTTR-dependent genes. Genes associated with 13 of the type three secretion system (T3SS), two of flagellar apparatus, ABC transporters, and transcription factors were expressed at lower levels in the NN14 mutant compared to the wild type. Conversely, tssH and hcp, type six secretion system (T6SS)-related genes, showed higher expression in NN14. Furthermore, these differences in gene expression were observed in minimal medium, but not in nutrient-rich medium, suggesting that LTTR acts as a global regulator responsive to nutrient conditions. Additionally, LTTR activated the expression of T3SS-related genes during Pcal infection. We also demonstrated that NN14 showed a reduced ability to induce hypersensitive reaction (HR) cell death in non-host plants. Collectively, these results suggest that LTTR contributes to Pcal virulence by regulating T3SS in response to environmental changes.

Evaluating the Impact of Environmental Factors on Bacterial Populations in Riverine, Estuarine, and Coastal Sediments

Aquatic ecosystems, including rivers, estuaries, and coastal environments, are crucial for maintaining biodiversity, regulating nutrient cycles, and supporting human livelihoods. However, these ecosystems are increasingly being threatened by urbanization, making it essential to understand their microbial communities and their ecological roles. This study employed high-throughput 16S rRNA gene sequencing to characterize the bacterial communities within the riverine, estuarine, and coastal sediments of Adyar Creek, Chennai, India. Proteobacteria were the dominant phylum across most samples, with proportions ranging from 39.65% to 72.09%. Notably, the estuarine environment exhibited a distinct taxonomic profile characterized by a significant abundance of Firmicutes (47.09% of the bacterial population). Distinct bacterial classes were observed across sediment types: Alphaproteobacteria (30.07–34.32%) in riverine sediments, Bacilli dominated estuarine sediments (40.17%), and Gammaproteobacteria (15.71–51.94%) in coastal sediments. The most significant environmental factors influencing the bacterial community composition across these samples were pH, salinity, phosphate, and nitrate. LEfSe (Linear discriminant analysis Effect Size) analysis identified specific genera within the estuary, including Bacillus (20.26%), unclassified_Paenibacillus (12.87%), Clostridium (3.81%), Gailella (3.17%), Paenibacillus (3.02%), Massilia (1.70%), Paraburkholderia (1.42%), and Pantoea (1.15%), as potential biomarkers for habitat health. Functional analysis revealed an elevated expression of the genes associated with ABC transporters and carbon metabolism in the estuary, suggesting a heightened nutrient cycling capacity. Furthermore, co-occurrence network analysis indicated that bacterial communities exhibit a strong modular structure with complex species interactions across the three sediment types. These findings highlight bacterial communities’ critical role and the key drivers in estuarine ecosystems, establishing a baseline for further investigations into the functional ecology of these vulnerable ecosystems.

Alterations of gut microbiota and metabolome are associated with primary nephrotic syndrome in children

BackgroundPrimary nephrotic syndrome (PNS) is a common glomerular disease in children. Dysbiosis of gut microbiota acts as a cause of Treg abnormalities. However, the intestinal metabolic impact of PNS with children remains poorly understood. This study aims to investigate the dynamic changes of gut microbiota and it’s metabolism in children with PNS.MethodsFecal and peripheral blood samples were separately collected from patients with initial diagnosis of PNS (PNS_In group), recurrence of PNS (PNS_Re group), and healthy controls (HCs group). The fecal samples were subjected to the microbiome and metabolome by the multi-omics analysis. Additionally, the peripheral blood samples were collected and associated inflammatory indicators were determined.ResultsWe found that in PNS_In group, lipopolysaccharide (LPS), pro-inflammatory interleukin (IL)-6, IL-17A, IL-23p19, and IL-1β were significantly increased compared with those in HCs group. However, these abnormalities were dramatically reversed in PNS_Re group treated with prednisone acetate. Moreover, the crucial Treg/Th17 axis in PNS inflammation was also proved to be discriminated between PNS and HCs. Gut microbial dysbiosis was identified in PNS_In and PNS_Re patients. At the genus level, compared to HCs group, the abundance of Faecalibacterium notably changed in PNS_In and PNS_Re groups, showing negatively correlated with inflammatory factors. Moreover, the fecal metabolome of PNS_In and PNS_Re remarkably altered with the major impacts in the metabolism of phenylalanine, ABC transporters, arginine and proline.ConclusionThe dynamic changes of gut microbiota and associated metabolites are closely correlated with initial period and recurrence of PNS in children via probably regulating inflammatory Th17/Treg axis, which may potentially provide novel targets for the control of the disease.Clinical trial numberNot applicable.

Cryptosporidium parvum infection alters the intestinal mucosa transcriptome in neonatal calves: impacts on epithelial barriers and transcellular transport systems

IntroductionCryptosporidium parvum (C. parvum) is the most prevalent enteric protozoan parasite causing infectious diarrhea in neonatal calves worldwide with a direct negative impact on their health and welfare. This study utilized next-generation sequencing (NGS) to deepen our understanding of intestinal epithelial barriers and transport mechanisms in the pathophysiology of infectious diarrhea in neonatal calves, which could potentially unveil novel solutions for treatment.MethodsAt day 1 of life, male Holstein-Friesian calves were either orally infected (n = 5) or not (control group, n = 5) with C. parvum oocysts (in-house strain LE-01-Cp-15). On day 8 after infection, calves were slaughtered and jejunum mucosa samples were taken. The RNA was extracted from collected samples and subjected to sequencing. Differentially expressed genes (DEG) between the infected and CTRL groups were assessed using DESeq2 at a false discovery rate < 0.05 and used for gene ontology (GO) and pathway enrichment analysis in Cytoscape (v3.9.1).Results and discussionTo study the pathophysiology of infectious diarrhea on intestinal permeability, 459 genes related to epithelial cell barrier integrity and paracellular and transmembrane transport systems were selected from 12,908 identified genes in mucus. Among, there were 61 increased and 109 decreased gene transcripts belonged to adhesion molecules (e.g. ADGRD1 and VCAM1), ATP-binding cassette (ABC, e.g. ABCC2 and ABCD1) and solute carrier (SLC, e.g. SLC28A2 and SLC38A3) transporters, and ion channels (e.g. KCNJ15). Our results suggest deregulation of cellular junctions and thus a possibly increased intestinal permeability, whereas deregulation of ABC and SLC transporters and ion channels may influence the absorption/secretion of amino acids, carbohydrates, fats, and organic compounds, as well as acid-based balance and osmotic hemostasis. Besides pathogen-induced gene expression alterations, part of the DEG may have been triggered or consequently affected by inflammatory mechanisms. The study provided a deeper understanding of the pathophysiology of infectious diarrhea in neonatal calves and the host-pathogen interactions at the transcript level. For further studies with a particular focus on the transport system, these results could lead to a new approach to elucidating pathophysiological regulatory mechanisms.

Coptisine enhances the sensitivity of chemoresistant breast cancer cells by inhibiting the function and expression of ABC transporters

BackgroundMultidrug resistance (MDR), mainly caused by ATP-binding cassette transporters (ABCTs) efflux, makes it difficult for many anticancer drugs to treat breast cancer (BC). Phytochemicals can reverse cancer’s MDR by modifying ABC transporter expression and function, as well as working synergistically with anticancer drugs to target other molecules. The reversal effect of the isoquinoline alkaloid coptisine (COP) was assessed on four breast cell lines; Two sensitive MCF-7 cell lines with positive estrogen, androgen, progesterone, and glucocorticoid receptors, as well as MDB-MB-231 cells with negative estrogen, progesterone, and HER2 receptors, and two doxorubicin-resistant cell lines, MCF-7/ADR and MDB-MB-231/ADR.MethodsThe cytotoxicity of COP and its ability to improve doxorubicin (DOX) cytotoxicity were assessed using the MTT assay. The effectiveness of COP in reversing DOX resistance was evaluated by calculating resistance ratio (RR) values, combination index (CI), and isobologram (IB). The inhibitory effect of COP on ABCT efflux function in comparison to verapamil (VER) was evaluated by measuring the cellular accumulation of Rho123 using flow cytometry. The impact of COP, either alone or in combination with DOX, on the gene expression of ABCTs (P-gp/MDR1, BCRP, and MRP1) of investigated cell lines was assessed by RT-PCR.ResultsThe COP showed modest cytotoxicity on the examined cell lines. In MCF-7/ADR and MDA-MB-231/ADR cells, COP (31 μM) enhanced DOX cytotoxicity with CI (0.77 and 0.75), RR (2.58 and 3.33), and IB suggesting synergism. COP significantly inhibits ABCT function in resistant BC cell lines, increases Rho123 accumulation, and decreases efflux more than VER; 2.1 and 1.2-fold, respectively. The combination of COP and DOX had a strong inhibitory effect on ABCT function (3.1 and 3.9 times VER, P< 0.001) and downregulated the genes and protein expression of ABCT.ConclusionCOP reversed ABCT-mediated multidrug resistance in vitro, indicating its potential as a multidrug resistance-reversing agent in cancer chemotherapy.

Impact of β-Carotene Enrichment on Carotenoid Composition and Gene Expression in Artemia Metanauplii

Background: Carotenoids play essential nutritional and physiological roles in aquatic animals. Since aquatic species cannot synthesize carotenoids de novo, they must obtain these compounds from their diet to meet the physiological and adaptive requirements needed in specific aquaculture stages and conditions. Carotenoid supplementation in Artemia represents a promising strategy to enhance pigmentation, health, and growth in aquaculture species, particularly in larvae and other early developmental stages. Methods: In this study, a β-carotene enrichment process was applied to Artemia metanauplii to investigate the biological fate and potential effects of β-carotene. Results: The results indicated significant β-carotene uptake by Artemia, with peak levels observed at 12 h. Alongside β-carotene, two xanthophylls—canthaxanthin and echinenone—were detected in Artemia, each exhibiting distinct patterns during the enrichment and subsequent depletion phases. The transcriptome analysis identified 2705 differentially expressed genes (DEGs), offering valuable insights into gene functions associated with carotenoid absorption, metabolism, and antioxidant mechanisms. The findings suggest that β-carotene enrichment enhances metabolic activity and energy pathways, supporting the physiological functions of Artemia. Notably, unlike other crustaceans, Artemia lack certain enzymes necessary for converting β-carotene into astaxanthin, restricting them to producing keto-carotenoids like canthaxanthin. Furthermore, the study highlights the upregulation of genes encoding lipid transport proteins, such as CD36 and ABC transporters, which may contribute to carotenoid absorption in Artemia. Additional functional insights are provided by the gene BCO2, which regulates pigmentation by preventing excessive carotenoid accumulation, along with ketolase and hydroxylase enzymes in carotenoid metabolic pathways. Conclusions: This research advances our understanding of carotenoid metabolism in crustaceans, with potential implications for aquaculture nutrition and feed formulation.

Environmental Changes Driving Shifts in the Structure and Functional Properties of the Symbiotic Microbiota of Daphnia

Symbiotic microbiota significantly influence the development, physiology, and behavior of their hosts, and therefore, they are widely studied. However, very few studies have investigated the changes in symbiotic microbiota across generations. Daphnia magna originating from the Qinghai–Tibetan Plateau were cultured through seven generations in our laboratory, and the symbiotic microbiota of D. magna were sequenced using a 16S rRNA amplicon to analyze changes in the structure and functional properties of the symbiotic microbiota of D. magna from a harsh environment to an ideal environment. We detected substantial changes in the symbiotic microbiota of D. magna across generations. For example, the genus Nevskia, a member of the gamma-subclass Proteobacteria, had the highest abundance in the first generation (G1), followed by a decrease in abundance in the fourth (G4) and seventh (G7) generations. The gene functions of the microbiota in different generations of D. magna also changed significantly. The fourth generation was mainly rich in fatty acyl-CoA synthase, acetyl-CoA acyltransferase, phosphoglycerol phosphatase, etc. The seventh generation was mainly rich in osmotic enzyme protein and ATP-binding protein of the ABC transport system. This study confirms that the alterations in the structure and functional properties of the symbiotic microbiota of D. magna under changing environments are typical responses of D. magna to environmental changes.

GLP-1RA improves diabetic renal injury by alleviating glomerular endothelial cells pyrotosis via RXRα/circ8411/miR-23a-5p/ABCA1 pathway.

Lipotoxicity has been implicated in diabetic kidney disease (DKD). However, the role of high glucose levels in DKD and the underlying renal protective mechanisms of GLP-1 receptor agonists (GLP-1RAs) remain unclear. To investigate cholesterol accumulation, pyroptosis in glomerular endothelial cells (GEnCs), and the renal protective mechanisms of GLP-1RAs, we used various techniques, including RT-qPCR, Oil Red O staining, Western blotting, lactate dehydrogenase (LDH) activity assays, circRNA microarrays, bioinformatics analysis, gain and loss-of-function experiments, rescue experiments, and luciferase assays. Additionally, in vivo experiments were conducted using C57BL/6J and ApoE-deficient (ApoE-/-) mice. GEnCs exposed to high glucose exhibited reduced cholesterol efflux, which was accompanied by downregulation of ATP-binding cassette transporter A1 (ABCA1) expression, cholesterol accumulation, and pyroptosis. Circ8411 was identified as a regulator of ABCA1, inhibiting miR-23a-5p through its binding to the 3'UTR. Additionally, higher glucose levels decreased circ8411 expression by inhibiting RXRα. GLP-1RAs effectively reduced cholesterol accumulation and cell pyroptosis by targeting the RXRα/circ8411/miR-23a-5p/ABCA1 pathway. In diabetic ApoE-/- mice, renal structure and function were impaired, with resulted in increased cholesterol accumulation and pyroptosis; however, GLP-1RAs treatment reversed these detrimental changes. These findings suggest that the RXRα/circ8411/miR-23a-5p/ABCA1 pathway mediates the contribution of high glucose to lipotoxic renal injury. Targeting this pathway may represent a potential therapeutic strategy for patients with DKD and hypercholesterolemia. Moreover, GLP-1RAs may provide renal protective effects by activating this pathway.

The ABC transporter Opp imports reduced glutathione, while Gsi imports glutathione disulfide in Escherichia coli

Glutathione is the major thiol-based antioxidant in a wide variety of biological systems, ranging from bacteria to eukaryotes. As a redox couple, consisting of reduced glutathione (GSH) and its oxidized form, glutathione disulfide (GSSG), it is crucial for the maintenance of the cellular redox balance. Glutathione transport out of and into cellular compartments and the extracellular space is a determinant of the thiol-disulfide redox state of the organelles and bodily fluids in question, but is currently not well understood. Here we use the genetically-encoded, glutathione-measuring redox probe Grx1-roGFP2 to comprehensively elucidate the import of extracellular glutathione into the cytoplasm of the model organism Escherichia coli. The elimination of only two ATP-Binding Cassette (ABC) transporter systems, Gsi and Opp, completely abrogates glutathione import into E. coli’s cytoplasm, both in its reduced and oxidized form. The lack of only one of them, Gsi, completely prevents import of GSSG, while the lack of the other, Opp, substantially retards the uptake of reduced glutathione (GSH).

Amelioration of arsenic-induced hepatic injury via sulfated glycosaminoglycan from swim bladder: Modulation of Nrf2 pathway and amino acid metabolism.

Arsenic, a known environmental pollutant with a carcinogenic risk, is associated with chronic liver toxicity. Prebiotic regulation represents an emerging dietary strategy to alleviate arsenic-induced hepatotoxicity; however, research in this area remains limited. This study employed sulfated swim bladder glycosaminoglycan (SBSG), a potential prebiotic, to assess its efficacy in mitigating arsenic-induced liver injury. In basic indicators, SBSG resisted oxidative stress by down-regulating AST, ALT, MDA, and MPO, up-regulating antioxidants (T-SOD, GSH, and GSH-px), and ameliorating histopathological damage. RT-qPCR analysis revealed that SBSG could regulate the Nrf2 signaling pathway and affect the expression of o genes related to ferroptosis and detoxification. The expression of protein further verified that SBSG could play an antioxidant and detoxifying role as an Nrf2 activator. Non-targeted metabolomics results demonstrated that SBSG primarily addresses metabolic disorders by up-regulating D-amino acid metabolism, ABC transporter, and alanine, aspartate and glutamate metabolism. Correlation analysis suggests that SBSG alleviates arsenic-induced liver oxidative damage through mechanisms linked to the Nrf2 pathway and amino acid metabolism. This study provided a research basis for expanding the dietary strategy to reduce arsenic induced toxicity.

Combined transcriptome and metabolome analysis reveals the mechanism of high nitrite tolerance in freshwater mussel Anodonta woodiana

Nitrite contamination and stress on aquatic organisms are increasingly emphasized in freshwater ecosystems. Freshwater bivalves exhibit high tolerance to nitrite; however, the underlying mechanism is unknown. Accordingly, this study investigated the tolerance mechanism of the globally occurring freshwater bivalve Anodonta woodiana. A. woodiana were exposed to nominal concentrations of 0, 250, 500, 1000, 2000, and 4000 mg/L nitrite for 96 h to calculate the 96-h median lethal concentration (96-h LC50). A combined transcriptome and metabolome analysis of the hemolymph (the most vital component of the bivalve immune system) was performed after exposing A. woodiana to 300 mg/L nitrite (approximately half the 96-h LC50) for 96 h. The 96-h LC50 of nitrite in A. woodiana was 618.7 mg/L. Transcriptome analysis identified 5600 differentially expressed genes (DEGs) primarily related to ribosomes, lysosomes, DNA replication, and nucleotide excision repair. Metabolome analysis identified 216 differentially expressed metabolites (DEMs) primarily involved in biosynthesis of amino acids, 2-oxocarboxylic acid metabolism, protein digestion and absorption, aminoacyl-tRNA biosynthesis, nucleotide metabolism, ABC transporters, and valine, leucine and isoleucine degradation. Combined transcriptome and metabolome analysis revealed that DEGs and DEMs were primarily associated with nucleotide (purine and pyrimidine) and amino acid metabolism (including aminoacyl-tRNA biosynthesis, cysteine and methionine metabolism, arginine and proline metabolism, and valine, leucine and isoleucine degradation) as well as the immune system (necroptosis and glutathione metabolism). This study is the first to describe the high tolerance of A. woodiana to nitrite and elucidate the molecular mechanisms underlying high nitrite tolerance in mussels.

Cadmium alters the cellular metabolome of human ovarian granulosa cells

Cadmium (Cd) is a toxic heavy metal that has been extensively implicated in disordered folliculogenesis, but the mechanisms underlying the ovarian toxicity of Cd remain to be explored fully. Granulosa cells are key players in ovarian follicular development and are the primary cells affected by Cd exposure-induced damage and dysfunction. In this study, we investigated how various levels of exposure of Cd (3 and 10 μM) to human granulosa cells (KGN cells) impacted the metabolism of the KGN cells utilizing a non-targeted metabolomics methodology. In vitro cell experiments revealed that Cd exposure dose-dependently diminished the viability of KGN cells. Metabolomics analysis revealed the presence of 296 (182 elevated and 114 reduced) and 397 (244 elevated and 153 reduced) differentially expressed metabolites after exposure to 3 and 10 μM, respectively. Cd exposure was found to significantly enrich nucleotide metabolism, sphingolipid metabolism, and ABC transporters in both groups. Although amino acid metabolic pathways exhibited significant enrichment across all groups, only glutathione, cysteine, and methionine metabolism were notably enriched in KGN cells exposed to 3 μM Cd, while glutathione and tryptophan metabolism were significantly enriched in the 10 μM Cd exposure cohort. The outcomes of this study provide mechanistic clues for elucidating Cd’s cytotoxic impact on granulosa cells, and deepen our understanding of the ovarian toxicity of Cd.

Integrated inactivation of Microcystis aeruginosa and degradation of microcystin-LR by direct current glow discharge plasma in liquid-phase: Mechanisms and cell deactivation process

The frequent occurrence of blooms of Microcystis aeruginosa (M. aeruginosa) and the subsequent release of microcystin-LR (MC-LR) in eutrophic waters pose a serious threat to aquatic ecosystems. This study investigated the optimal conditions for inactivating M. aeruginosa and the degrading MC-LR using direct current glow discharge plasma in liquid phase (DC-LGDP), analyzed the potential inactivation mechanisms and the cell deactivation process of M. aeruginosa. The results showed that DC-LGDP generated reactive species (i.e., •OH, 1O2, and H2O2), active Cl and electroporation effect collectively contributed to inactivation of M. aeruginosa and degradation of MC-LR. The 97.07% inactivation efficiency of M. aeruginosa and 94.98% degradation rate of MC-LR were achieved with higher energy yield and without generating nitrogen oxides. Meanwhile, DC-LGDP destroyed the cell integrity, eliminated their antioxidant capacity and reduced the content of photosynthetic pigments. The transcriptome analysis indicated that the transcripts of genes related to photosynthesis, ribosome biosynthesis, ABC transporters, and nitrogen metabolism pathway in M. aeruginosa were altered by DC-LGDP. This study provides insights into the inactivation of M. aeruginosa by DC-LGDP, while elucidating the potential inactivation mechanisms and the cell deactivation process involved. It may be important for the eco-friendly inactivation of M. aeruginosa blooms in natural water bodies. Environmental implicationsThe frequent occurrence of blooms of M. aeruginosa and the subsequent release of MC-LR in eutrophic waters are a global issue. Although glow discharge plasma technologies have been applied for the inactivation of M. aeruginosa, limitations such as low energy yield and additional nitrous oxide production remain. This study aims to explore the optimal conditions for inactivating M. aeruginosa and degrading MC-LR using direct current glow discharge plasma in liquid phase (DC-LGDP), as well as to analyze the inactivation mechanisms and the cell deactivation process of M. aeruginosa. Our results show that the 97.07% inactivation efficiency of M. aeruginosa by DC-LGDP is achieved at higher energy yield and without the generation of nitrogen oxides. The inactivation of M. aeruginosa and degradation of MC-LR are involved in the reactive species generated by DC-LGDP and electroporation effect. These factors induce oxidative stress damaging the cell integrity, impairing the photosynthesis and ribosome biosynthesis pathways of M. aeruginosa, ultimately leading to the cell deactivation of M. aeruginosa. Our findings provide insights into the effective inactivation of M. aeruginosa and the degradation of MC-LR by DC-LGDP with lower energy consumption and environmental friendliness. Thus, it may be important for the inactivation of M. aeruginosa blooms in natural water bodies.

Identification of SNPs and INDELS associated with duck egg quality traits through a genome-wide association analysis

Egg quality traits are economically important in the poultry industry. To explore the genetic architecture and identify potential candidate genes, a genome-wide association study (GWAS) was performed for 13 egg quality traits using data from whole-genome sequencing of 299 Longyan Shan-ma female ducks, including 12 quantitative traits and one qualitative trait, eggshell color (ESC; white, light green, green). From estimation of pedigree genetic parameters, heritability (h2) ranged from 0.022 to 0.996 for the 12 quantitative traits, with the highest h2 (0.996) for eggshell color a* value (ESCA) and the lowest h2 (0.022) for egg yolk percentage relative to EW. A total of 8,874 single nucleotide polymorphism (SNP)-based significant associations (1.0 × 10-6) and 247 insertion-deletion (indel)-based significant associations (1.00 × 10-5) were identified, including 5,980 SNPs and 159 indel markers. From 5,924 SNPs and 143 indels associated with ESC traits, 181 potential candidate genes were identified, and most significant SNPs and indels (P < 1.0 × 10-20) were located at 1.86 Mb (44.29-46.15 Mb) on chromosome 4. The top SNP (chr4:45325309:C>A; P = 7.97 × 10-43) and the top indel (chr4:45299595:delTTCCACTCCAC; P = 4.20 × 10-36) for the ESC a* value were within two known ESC candidate genes; ATP-binding cassette subfamily G member 2 (ABCG2) and protein kinase cGMP-dependent 2 (PRKG2). Of 56 SNPs and 16 indels associated with other egg quality traits, 46 potential candidate genes were identified including synapse differentiation-inducing 1-like (SYNDIG1L) for EW, and core histone macro-H2A.1 (LOC101795967) and neurogenin 1 (NEUROG1) for egg shape index; and four genes including collagen type VI alpha 3 chain (COL6A3), lysine demethylase 7A (KDM7A), LOC101802169, and sperm-associated antigen 16 (SPAG16) for egg yolk weight and the percentage of yolk to total egg weight. Of the 46 genes, the molecular functions of 22 are related to protein binding, indicating important roles in the formation of egg quality traits. Our findings provide new insight into the genetic basis of egg quality traits in ducks.

Combining ultrasound technology with targeted fucoidan/arginine-gelatin nanoparticles loaded with doxorubicin to enhance therapeutic efficacy and modulate bioeffects in drug-resistant triple-negative breast cancer.

Triple-negative breast cancer (TNBC) presents formidable challenges due to its aggressive nature and high recurrence rates, compounded by the involvement of epithelial-mesenchymal transition (EMT) in its progression and metastasis. Standard chemotherapy, which typically employs doxorubicin (DOX), remains a primary treatment approach. However, multidrug resistance (MDR) mechanisms, which include ATP-binding cassette transporters and EMT, contribute to treatment failures. Ultrasound has emerged as a promising modality among the various strategies explored to address MDR in TNBC. It serves as a diagnostic tool and holds therapeutic potential by inducing various biological effects depending on the exposure level. Targeted nanoparticles offer a means to enhance drug delivery efficiency. Our study aims to advance ultrasound technology combined with biocompatible nanoparticles using simplified preparation methods to improve treatment outcomes for drug-resistant TNBC. In particular, employing DOX-loaded fucoidan/arginine-gelatin nanoparticles facilitated the targeted delivery of chemotherapy drugs to tumors by effectively interacting with P-selectin, resulting in tumor growth inhibition. Furthermore, these nanoparticles mitigated MDR and EMT, particularly when combined with ultrasound treatment. This integrated approach of nanoparticle delivery with ultrasonography opens up a promising and innovative avenue for clinical cancer research.

Febuxostat enhances the efficacy of dasatinib by inhibiting ATP-binding cassette subfamily G member 2 (ABCG2) in chronic myeloid leukemia cells.

Dasatinib is a second-generation breakpoint cluster region-abelson 1 (BCR::ABL1) tyrosine kinase inhibitor (TKI) used for the treatment of chronic myeloid leukemia (CML). Overexpression of ATP-binding cassette subfamily G member 2 (ABCG2) in CML cells contributes to dasatinib resistance and poor chemotherapeutic responses. Considering that the xanthine oxidase inhibitor febuxostat has anti-ABCG2 activity, febuxostat may enhance the efficacy of dasatinib. However, the mechanism of action of febuxostat and its effects on the efficacy and safety of dasatinib in patients with CML are unknown. Therefore, this study aimed to retrospectively investigate the clinical impact of concomitant febuxostat on the efficacy of dasatinib in 65 patients with CML. Moreover, its underlying mechanism was explored in vitro using an ABCG2-overexpressing CML cell line (K562-ABCG2 cells). The retrospective study revealed that the achievement ratios of early molecular response at three months and major molecular response at 12 months after dasatinib treatment in patients with febuxostat were significantly higher than those in patients without febuxostat (91 % vs. 70 %, p = 0.034, 86 % vs. 53 %, p = 0.013, respectively). In vitro studies showed that febuxostat significantly decreased cell viability and increased the residual dasatinib concentration in dasatinib-treated K562-ABCG2 cells. Moreover, phosphorylated BCR::ABL1 levels in dasatinib-treated K562-ABCG2 cells were significantly decreased by febuxostat. Overall, concomitant febuxostat enhanced the efficacy of dasatinib in patients with CML. This was achieved partially by inhibition of ABCG2-mediated excretion of dasatinib from CML cells. Therefore, these findings provide important insights for improving CML treatment and overcoming TKI resistance.