Search Results (10,052)

Ethylene is a multifunctional phytohormone that regulates plant growth, development, and responses to abiotic/biotic stresses. RTE1 (Reversion-To-Ethylene Sensitivity1) acts as a negative regulator of the ethylene responses in Arabidopsis by positively regulating ethylene receptor ETR1. However, the role of RTE genes in sweet potato (Ipomoea batatas), an import food crop worldwide, remains largely unknown, particularly their involvement in abiotic stress adaptation. In this study, we identified 23 RTE genes in sweet potato, distributed across 21 chromosomes and one scaffold BrgTig00017944. The phylogenetic analysis divided them into two groups, the RTE1 group and RTH (RTE1-Homolog) group. Synteny analysis revealed that whole genome duplication (WGD) was the major force of expansion of the IbRTE gene family. Multiple cis-acting elements responsive to hormones and stress were found in the promoter region of IbRTE genes. The transcriptome expression profiling showed that the majority of IbRTEs have tissue-specific and differential expression under drought and salt stresses. Meanwhile, the qRT–PCR results showed that the 14 representatives IbRTEs have differential expression profilings under salt (NaCl) and drought (PEG) treatments. These findings suggest that the IbRTE genes may be involved in sweet potato’s adaptive responses to salt and drought, providing a valuable foundation for further functional studies. Full article

Background/Objectives: Advanced 3D cell culture techniques enhance the physiological relevance of in vitro models, while supporting the 3Rs principles (Reduction, Refinement, and Replacement) of animal experimentation. In this context, 3D collagen-based systems mimic key extracellular matrix properties, enabling more accurate cellular organization and phenotype. However, changes in culture dimensionality can affect RT-qPCR reference gene stability, underscoring the need for careful validation when combining 2D and 3D systems. Methods: AML12 cells were cultured for 7 days under different 2D and collagen-based 3D conditions. The expression stability of nine candidate housekeeping genes was systematically evaluated using established algorithms (BestKeeper, NormFinder, geNorm, RefFinder, and ΔCt method), followed by inter-group statistical and correlation analyses of raw Ct values. Albumin gene expression was used as a target gene. Results: Although all candidate genes initially met acceptable variability thresholds, a stepwise, exclusion-based analysis revealed distinct performance differences. Hprt, Ppia, and Actb emerged as the most stable, showing no intra-group variability or interaction with Albumin expression. Nevertheless, Ywhaz and Rplp0, despite their high stability, were compromised by significant correlation with Albumin. Furthermore, Ywhaz showed significant downregulation under 3D culture conditions. B2M, Gapdh, 18S, and Hmbs exhibited increased variability, likely reflecting metabolic and microenvironmental heterogeneity associated with prolonged 2D cultivation of AML12 cells. Conclusions: Overall, this study highlights the importance of context-dependent, exclusion-based reference gene validation when comparing 2D and 3D models, and demonstrates a new approach for reliable gene expression normalization in complex in vitro culture systems. Full article

Background: Diabetic nephropathy (DN) is largely driven by transforming growth factor-β1 (TGF-β1)-mediated fibrosis. Clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) ribonucleoprotein (RNP) complexes offer precise gene disruption, yet effective non-viral delivery remains a challenge. This study developed cationic lipid-based hybrid nanostructured lipid carriers (NLCs) for intracellular delivery of TGFB1-targeting RNP as an early-stage platform for DN gene modulation. Methods: A single-guide RNA (sgRNA) targeting human TGFB1 was assembled with Cas9 protein (1:1 and 1:2 molar ratios). Hybrid NLCs comprising squalene, glyceryl trimyristate, and the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) were formulated via optimized emulsification–sonication to achieve sub-100 nm particles. Physicochemical properties, including polydispersity index (PDI), were assessed via dynamic light scattering (DLS), while silencing efficacy in HEK293T cells was quantified using quantitative reverse transcription PCR (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). Results: Optimized NLCs achieved hydrodynamic diameters of 65–99 nm (PDI < 0.5) with successful RNP complexation. The 1:2 Cas9:sgRNA formulation produced the strongest gene-editing response, reducing TGFB1 mRNA by 67% (p < 0.01) compared with 39% for the 1:1 ratio. This translated to a significant reduction in TGF-β1 protein (p < 0.05) within 24 h. Conclusions: DOTAP-based hybrid NLCs enable efficient delivery of CRISPR–Cas9 RNP and achieve significant suppression of TGFB1 expression at both transcriptional and protein levels. These findings establish a promising non-viral platform for upstream modulation of profibrotic signaling in DN and support further evaluation in kidney-derived cells and in vivo renal models. Full article

This study was conducted to elucidate the molecular and metabolic differences in ileal development according to birth weight in neonatal piglets. A total of 126 neonatal piglets born from Yorkshire × Landrace × Duroc crossbred sows were used, and the top 5% (H group, 1.77 ± 0.02 kg) and bottom 5% (L group, 0.72 ± 0.03 kg) of birth weights were selected for analysis. Ileal tissues were collected for transcriptomic (RNA-seq) and targeted metabolomic (GC–MS) analyses, and selected genes were validated using RT-qPCR. A total of 112 differentially expressed genes (DEGs) were identified, among which RFC3, PCNA, MCM3, MCM10, AURKA, AURKB, CCNB2, CCNA2, CCNF, and SI were significantly upregulated in the H group (p < 0.05). These genes were mainly involved in pathways related to DNA replication, cell division, and nutrient digestion and absorption. In addition, metabolomic analysis revealed that pyruvic acid concentrations were significantly higher in the H group (p < 0.05), indicating the activation of energy metabolic pathways. These results indicate that high-birth-weight piglets possess a genetic foundation for enhanced cellular proliferation and energy metabolism, and they further highlight potential molecular targets for improving growth performance and intestinal development in low-birth-weight piglets. Full article

Background: Madariaga virus (MADV), formerly known as the South American variant of Eastern Equine Encephalitis virus (EEEV), is an alphavirus that belongs to the Togaviridae family and has been periodically infecting equids in Venezuela since its first identification in 1975. This study reports the isolation and molecular characterization of MADV isolated from a horse in December 2024 in the context of MADV cases reported in Venezuela. Methods: Antibodies to the rabies virus were detected by indirect immunofluorescence, and to the Equine Infectious Anemia virus (EIAV) by passive immunodiffusion. MADV RNA was detected by qRT-PCR. The sequence of the complete viral genome was obtained by next-generation sequencing. Results: The sequence of this virus was highly similar to that of the only human case of MADV reported in the country in 2016, as well as to a sequence of a virus isolated from a horse in Colombia in 2002. The horse was found to be co-infected with EIAV. Conclusions: The continuous circulation of MADV in Venezuela warrants reinforcing the preventive measures against these alphaviruses, which ignore borders, and may cause important animal and human health concerns. Full article

Actinidia deliciosa is a globally important economic fruit crop, and its fruit quality and yield are profoundly influenced by light and environmental conditions. Sucrose phosphate synthase (SPS), a key rate-limiting enzyme in the sucrose biosynthesis pathway, plays a central role in regulating carbon metabolism and sucrose accumulation in plants. However, comprehensive studies of the SPS gene family in A. deliciosa are still lacking, particularly regarding its expression in response to different light qualities. In this study, genome-wide identification of the SPS gene family in A. deliciosa was conducted using bioinformatics approaches. A total of 31 SPS genes were identified and named AdSPS1 to AdSPS31 on the basis of their chromosomal positions. The encoded proteins were predicted to be acidic, hydrophilic, and primarily localized in the chloroplast. All the AdSPS proteins contained the conserved domains Sucrose_synth, Glyco_trans_1, and S6PP, indicating potential roles in sucrose metabolism. Phylogenetic analysis classified the 31 AdSPS members into three subfamilies, A, B, and C, comprising 20, 5, and 6 members, respectively. Collinearity analysis revealed extensive syntenic relationships among AdSPS genes across different chromosomes, suggesting that gene duplication events contributed to the expansion of this gene family. Promoter cis-acting element analysis revealed that light-responsive elements were the most abundant among all the detected elements in the upstream regions of the AdSPS genes, implying potential regulation by light signals. Different light qualities significantly affected the contents of sucrose, glucose, and fructose, as well as SPS activity in kiwifruit leaves, with the highest activity observed under the R3B1 (red–blue light 3:1) treatment. Spearman’s correlation analysis indicated that AdSPS3 was significantly negatively correlated with sucrose, fructose, glucose, and SPS activity, suggesting a potential role in negatively regulating sugar accumulation in kiwifruit leaves, whereas AdSPS12 showed positive correlations with these parameters, implying a role in promoting sucrose synthesis. To further explore the light response of the AdSPS genes, eight representative members were selected for qRT‒PCR analysis under red light, blue light, and combined red‒blue light treatments. These results demonstrated that light quality significantly influenced SPS gene expression. Specifically, AdSPS6 and AdSPS24 were highly responsive to R1B1 (1:1 red‒blue light), AdSPS9 was significantly upregulated under R6B1 (6:1 red‒blue light), AdSPS21 was strongly induced by blue light, and AdSPS12 expression was suppressed. This study systematically identified and analyzed the SPS gene family in A. deliciosa, revealing its structural characteristics and light-responsive expression patterns. These findings suggest that AdSPS genes may play important roles in light-regulated carbon metabolism. These results provide a theoretical foundation and valuable genetic resources for further elucidating the molecular mechanisms of sucrose metabolism and light signal transduction in kiwifruit. Full article

The anti-aging gene/protein Klotho (Kl), most present in kidneys, has been well studied in mice (mKl), but not in rats (rKl). This study investigated the renal rKl expression in male and female rats. Sex-related measurement of rKl-controlled electrolytes was performed in plasma/urine samples, as were tests on species differences in renal Kl expression (rats vs. mice). rKl mRNA/protein expression was studied by qRT-PCR/Western-blotting in renal total RNA/cell membranes and its localization by immunofluorescence microscopy. Urine/plasma ions (phosphate/total calcium) and macroelements (phosphorus/calcium) were measured biochemically and by ICP-MS, respectively. In rat kidneys, the rKl mRNA/protein was detected in the cortex, outer and inner stripe but not in the papilla, and was immunolocalized in the basolateral membrane of proximal tubules in the cortex and outer stripe, but not in the intercalating cells of the cortical distal tubules, whereas mKl was observed in the mouse kidney cortex but not the outer stripe. Female-dominant expression of renal rKl, affected by androgen’s inhibitory effect, may have contributed to the sex-related level of urine electrolytes, particularly phosphates. Renal mKl expression was male-dominant. Sex- and species-related differences in renal Kl expression may be relevant for the selection of the sex and/or the model organism in studies addressing aging/mineral homeostasis. Full article

The YABBY proteins are a type of transcription factor known to participate in the growth and development of plants. In this study, we analyzed a genome-wide identification of the YABBY gene family in watermelon (Citrullus Lanatus). A total of nine ClaYABBY genes were identified and classified into five subfamilies. Sequence and phylogenetic analyses revealed that segmental duplication contributed to the expansion of this family. Two paralogous gene pairs not only exhibited highly similar sequences but also had relatively consistent expression patterns. RT-qPCR results indicated that ClaYABBY genes exhibited specific expression in different tissues, and most members showed higher expression levels in reproductive organs. Among them, ClaYABBY2 maintained a consistently high expression throughout fruit development, suggesting its potential role in the stage of fruit development. Furthermore, under drought and salt stresses, the expressions of ClaYABBY2, 3, 4, and 9 were upregulated, indicating that they may be involved in abiotic stress in watermelon. This study provides insight for future research on the specific functions of the ClaYABBY gene in fruit development and environmental adaptation of watermelon. Full article

The silkworm, Bombyx mori, is an economically important insect severely impacted by Bombyx mori nucleopolyhedrovirus (BmNPV), a double-stranded DNA virus that causes substantial losses to the sericulture industry. To elucidate the mechanisms of BmNPV infection and replication, we performed transcriptome sequencing of BmN cells infected with a recombinant BmNPV at 12 and 24 h post infection (hpi). A total of 1136 differentially expressed genes (DEGs) were identified in the 12 hpi group, including 789 up-regulated and 347 down-regulated genes, while 5191 DEGs were detected at 24 hpi, including 2102 up-regulated and 3089 down-regulated genes. Functional annotation via GO and KEGG analyses highlighted the ECM–receptor interaction pathway as particularly significant. Furthermore, RT-qPCR results demonstrated that Map3k12 inhibits BmNPV replication. These findings lay the groundwork for further investigation into the molecular mechanisms of BmNPV infection and can be utilized for breeding dominant genes conferring resistance to nucleopolyhedrovirus in the silkworm. Full article

Highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b viruses spread efficiently via migratory wild birds and increasingly infect mammals. The leopard cat (Prionailurus bengalensis) is an endangered mesopredator in South Korea that frequents wetland–forest ecotones and overlaps with wild waterbirds, placing it at risk of exposure. We describe a fatal HPAI H5N1 infection in a free-ranging leopard cat detected through national wildlife surveillance during a period of widespread H5N1 activity in wild birds along the East Asian–Australasian Flyway. The animal showed acute neurological and respiratory signs and died shortly after rescue. H5 viral RNA was detected by RT-qPCR in all examined tissues, with the highest load in the brain, and infectious virus was isolated from the brain, bronchoalveolar lavage fluid, and nasal swab. Pathology revealed acute serofibrinous pneumonia, severe nonsuppurative meningoencephalitis, necrotizing vasculitis with thrombosis, and necrotizing enteritis with secondary mesenteritis. Immunohistochemistry demonstrated abundant viral antigen in nasal and olfactory epithelium, olfactory bulb, neurons, endothelial cells, and bronchial and bronchiolar epithelium, supporting combined olfactory and hematogenous dissemination. This clinicopathological description expands the spectrum of HPAI-associated lesions in felids and underscores the value of wild carnivores as bioindicators of avian influenza spillover in a One Health context. Full article

Background/Objectives: In all types of diabetes, elevated blood glucose levels cause pathological changes in skeletal muscle, primarily due to oxidative stress, mitochondrial dysfunction, and excessive production of reactive oxygen species (ROS). Regular exercise can help mitigate these effects; however, the underlying mechanisms, particularly those involving the mitochondrial permeability transition pore (mPTP), remain incompletely understood. This study aimed to explore the effects of aerobic exercise training (AET) on oxidative stress and the expression of mPTP components in the skeletal muscle of streptozotocin-induced diabetic rats. Methods: Male Wistar rats were randomly divided into three groups: Healthy Sedentary (H-SED), Diabetic Sedentary (D-SED), and Diabetic Exercise-trained (D-EXER); n = 6 per group. The D-EXER group performed AET (0° slope) 5 days/week for 8 weeks. After the intervention period, body weight and fasting blood glucose (FBG) levels were measured, and soleus muscles were collected and analyzed for oxidative stress biomarkers, Western blotting, and gene expression using qRT-PCR. Results: Following an 8-week intervention, AET reduced FBG concentrations. Accordingly, in the soleus muscles of the D-EXER group, ROS levels decreased, and redox balance was improved compared to the D-SED group. Exercise training reduced CypD and Casp9 mRNA expression and increased Bcl-2 mRNA expression, whereas Ant1 mRNA expression was only slightly altered. CypD protein expression was decreased in exercised diabetic rats, while VDAC1 protein and mRNA levels remained unchanged. In the D-EXER group, there were significant inverse correlations between CypD and Casp9 mRNA expression levels and glutathione redox state. Conclusions: The current study suggests that 8 weeks of AET, in addition to reducing hyperglycemia, may favorably influence oxidative balance and the expression of mPTP-related molecular components in diabetic skeletal muscle. Full article

Lateral Organ Boundaries Domain (LBD) proteins are plant-specific transcription factors characterized by a typical N-terminal LOB domain and are critical for plant growth, development, and stress response. Currently, LBD genes have been investigated in various plant species, but they have yet to be identified in Neolamarckia cadamba, known as a ‘miracle tree’ for its fast growth and acknowledged for its potential medicinal value in tropical and subtropical areas of Asia. In this study, a total of 65 NcLBD members were identified in N. cadamba by whole-genome bioinformatics analysis. Phylogenetic analysis revealed their classification into two clades with seven distinct groups, and their uneven distribution across 18 chromosomes, along with 6 tandem repeats and 58 segmental duplications. Furthermore, enrichment analysis of transcription factor binding motifs within NcLBD promoters identified the MYB-related and WRKY families exhibited the most significant enrichment in the NcLBD promoter. Protein interaction network analysis revealed potential interactions among NcLBD proteins, as well as their interactions with various transcription factors. RNA-seq and qRT-PCR analyses of NcLBDs transcript levels showed distinct expression patterns both across various tissues and under different hormone and abiotic stress conditions. Specifically, NcLBD3, NcLBD37, and NcLBD47 were highly expressed in vascular cells and induced by abiotic stress, including cold, drought, and salt, suggesting their significant role in the processes. In summary, our genome-wide analysis comprehensively identified and characterized LBD gene family in N. cadamba, laying a solid foundation for further elucidating the biological functions of NcLBD genes. Full article

Background: 5-Azacytidine (5-Aza) is a clinically important DNMT inhibitor with the potential to modulate cardiac remodeling by epigenetically reprogramming human cardiac fibroblasts (HCFs). However, its clinical utility is limited by rapid hydrolytic degradation. Nanoparticle (NP) encapsulation offers a strategy to mitigate this instability. This study evaluated the physical and chemical stability of free 5-Aza and 5-Aza-loaded lipid nanoparticles (5-Aza-NP) under different storage temperatures and examined their effects on DNA methylation-related gene expression in HCFs. Methods: Hyaluronic acid-stabilized lipid NPs were prepared using a solvent displacement method. Particle size, polydispersity index (PDI), and zeta potential were monitored over four days at −20 °C, 4 °C, and 30 °C. Chemical stability was assessed using HPLC and first-order kinetic modeling. Functional activity was evaluated by treating HCFs with free 5-Aza or 5-Aza-NP stored for 96 h and measuring DNMT1, DNMT3A, and DNMT3B expression by RT-qPCR. Results: 5-Aza-NP remained physically stable at 4 °C, while −20 °C induced aggregation and 30 °C caused thermal variability. Free 5-Aza degraded rapidly at 30 °C (6.56% remaining at 72 h), whereas 5-Aza-NP preserved 11.54%. Kinetic modeling confirmed first-order degradation, with consistently longer half-lives for the NP formulation. Functionally, 5-Aza–NP preserved its ability to suppress DNMT1 expression following 96 h of storage at 4 °C, whereas free 5-Aza showed reduced activity. In contrast, DNMT3A and DNMT3B levels remained low and unchanged across all treatments. Conclusions: NP encapsulation enhances the physicochemical stability of 5-Aza and preserves its DNMT1-inhibitory activity, while DNMT3A/B remain unaffected. These findings support NP-based delivery as a promising strategy to stabilize labile epigenetic drugs such as 5-Aza. Full article

Background: Intestinal failure-associated liver disease (IFALD) is a serious complication in patients receiving parenteral nutrition, often exacerbated by inflammation, lipid overload, and oxidative stress. Nobiletin (NOB), a polymethoxylated flavone, is known for its anti-inflammatory and lipid-regulating properties. Methods: We employed an in vitro model using THLE-2 human hepatocytes and primary human cholangiocytes exposed to Intralipid (INT) and lipopolysaccharide (LPS) to simulate IFALD conditions. NOB was tested at non-toxic concentrations (10 and 25 µM) to assess its protective effects. MTT viability assays, multiplex bead-based immunoassays (MAGPIX), RT-qPCR, and Western blotting were used to evaluate changes in inflammation markers, gene expression, and protein signaling. Moreover, ALT and AST activities were used to assess hepatocellular injury. Results: NOB maintained high cell viability in THLE-2 hepatocytes and cholangiocytes, confirming its low cytotoxicity. NOB normalized ALT and AST activities in both tested cell lines, but the effect reached statistical significance only for ALT in cholangiocytes. Under IFALD-like conditions (LPS+INT), NOB significantly preserved metabolic activity in both cell types. In THLE-2 and cholangiocytes, NOB markedly reduced the phosphorylation of pro-inflammatory proteins JNK, NF-κB, and STAT3, indicating a broad inhibition of inflammatory signaling. Moreover, in THLE-2 cells, NOB upregulated lipid metabolism-related genes (PRKAA2, CYP7A1, and ABCA1) and decreased oxidative stress, thereby enhancing the nuclear translocation of Nrf2 and increasing SOD1 level, which supports the activation of antioxidant defenses. Conclusions: NOB exhibits hepatoprotective properties under IFALD-like conditions in vitro, likely through modulation of inflammation-related signaling and lipid metabolism pathways. Full article

Background: Ulcerative colitis (UC) is characterized by chronic mucosal inflammation, oxidative stress, and disruption of intestinal metabolic homeostasis. Immunomodulatory nutrients such as arginine, glutamine, and β-hydroxy β-methylbutyrate (HMB) have shown potential benefits; however, their combined molecular effects on UC remain insufficiently defined. Objective: To investigate the individual and combined effects of arginine, glutamine, and HMB on inflammatory and metabolic gene expression, oxidative stress markers, and histopathological outcomes in a dextran sulfate sodium (DSS)-induced colitis model. Methods: Female Sprague Dawley rats were assigned to six groups: control, DSS, DSS + arginine, DSS + glutamine, DSS + HMB, and DSS + mixture. Colitis was induced using 3% DSS. Colon tissues were examined histologically, serum MDA, MPO, and GSH levels were quantified, and mRNA expression of IL6, IL10, COX2, NOS2, ARG2, CCR1, and ALDH4A1 was measured by RT-qPCR. Pathway enrichment analyses were performed to interpret cytokine and metabolic network regulation. Results: DSS induced severe mucosal injury, elevated MDA and MPO, reduced GSH, and significantly increased IL6, COX2, NOS2, ARG2, and CCR1 expression. Glutamine demonstrated the strongest anti-inflammatory and antioxidant effects by decreasing IL6 and COX2 and restoring GSH. Arginine primarily modulated nitric oxide–related pathways, whereas HMB increased ALDH4A1 expression and metabolic adaptation. The combination treatment produced more balanced modulation across inflammatory, chemokine, and metabolic pathways, consistent with enrichment results highlighting cytokine signaling and amino acid metabolism. Histopathological improvement was greatest in the mixture group. Conclusions: Arginine, glutamine, and HMB ameliorate DSS-induced colitis through coordinated regulation of cytokine networks, oxidative stress responses, and metabolic pathways. Their combined use yields broader and more harmonized therapeutic effects than individual administration, supporting their potential as targeted immunonutritional strategies for UC. Rather than targeting a single inflammatory mediator, this study was designed to test whether combined immunonutrient supplementation could promote coordinated regulation of cytokine signaling, oxidative stress responses, and metabolic adaptation, thereby facilitating mucosal repair in experimental colitis. Full article