Search Results (865)

Enterovirus D68 (EV-D68) is a non-polio enterovirus that can cause a polio-like paralysis condition, acute flaccid myelitis (AFM). EV-D68-associated AFM cases waned in the US after 2018, and the reasons for this are unknown. It has recently been demonstrated that EV-D68 containing point mutations in viral structural proteins VP1 and VP3 resulted in decreased paralysis in different neonatal mouse models. However, phenotypes of these mutations in a human multicellular central nervous system (CNS) model are unknown. We hypothesize that mutations in VP1 and VP3 will similarly direct neurotropism in human spinal cord organoids (hSCOs). To investigate this, we recreated viruses with mutations in VP3 (I88V) or VP1 (L1I/N2D/T98A/E283K or L1P/V148A/K282R) and infected hSCOs. We found that VP3 I88V and VP1 L1I/N2D/T98A/E283K resulted in decreased titer and viral protein staining, consistent with attenuated neurovirulence in previously published murine models. We also found through immunofluorescence that VP1 L1P/V148/K282R mutations altered cellular tropism, primarily infecting glial cells rather than neuronal cells. When these mutations were combined, their effects on neurotropism were not additive. Sequence analysis of recently circulating EV-D68 strains reveals that VP3 I88 and VP1 E283 have remained the dominant amino acid residues since 2014, whereas VP1 sites 1, 2, and 98 have higher population diversity, indicating that these residues may be contributing to newly reduced neurovirulence after 2018. Full article

Metastatic pancreatic ductal adenocarcinoma (PDAC) is typically treated with fluorouracil, leucovorin, irinotecan, and oxaliplatin (FOLFIRINOX) or gemcitabine plus nab-paclitaxel (GnP), but the choice between regimens remains largely empirical. This narrative review summarizes biomarkers with potential to inform first-line selection in metastatic PDAC, emphasizing genomic and transcriptomic correlates of differential benefit. Recent head-to-head trials, particularly Pancreatic Adenocarcinoma Signature Stratification for Treatment (PASS-01) and GENERATE (Japan Clinical Oncology Group [JCOG] 1611), indicate that modified FOLFIRINOX (mFOLFIRINOX) is not uniformly superior to GnP, strengthening the rationale for biomarker-guided selection. The strongest evidence favoring platinum-based/FOLFIRINOX strategies involves homologous recombination repair deficiency (HRD), especially alterations in germline breast cancer gene 1/2 (BRCA1/2) or partner and localizer of BRCA2 (PALB2), as well as broader genomic scar signatures. Transcriptomic subtype and GATA-binding protein 6 (GATA6) expression are promising but remain unsettled because retrospective data favor classical/GATA6-high disease for FOLFIRINOX, whereas PASS-01 suggested better outcomes with GnP in classical tumors. Candidate biomarkers favoring GnP include high human equilibrative nucleoside transporter 1 (hENT1), low class III β-tubulin (TUBB3) expression, and exploratory phosphatidylinositol 3-kinase (PI3K)/KIT/NOTCH pathway mutation signals. Comprehensive molecular profiling also identifies actionable alterations that may redirect patients to targeted therapy or clinical trials rather than standard chemotherapy alone. Importantly, no biomarker has yet been prospectively validated in a biomarker-stratified randomized trial with regimen selection as the primary endpoint; all biomarker-regimen associations described in this review should therefore be considered hypothesis-generating rather than practice-defining. Nevertheless, the convergence of genomic, transcriptomic, and organoid-based approaches makes biologically informed first-line selection increasingly feasible in metastatic PDAC. Full article

Semaglutide, a long-acting glucagon-like peptide-1 receptor agonist (GLP-1RA), has transformed obesity and diabetes management. However, its expanding use among reproductive-age women raises concerns about potential effects on early placental development. We examined semaglutide’s impact on two human trophoblast cell lines: Swan71 (invasive extravillous) and BeWO (syncytiotrophoblast-like). Cells were treated with semaglutide (100 nM) for 24 h, and proliferation, viability, mitochondrial respiration, oxidative stress, signaling pathways, and invasiveness were evaluated. Semaglutide significantly reduced proliferation in Swan71 cells and increased it in BeWO cells, with no significant change in viability for Swan71 and a slight increase for BeWO. Western blot analysis revealed altered phosphorylation of key signaling proteins, including mTOR, p70S6K, 4EBP1, AKT, and ERK, as well as increased AMPK phosphorylation, indicating a shift toward catabolic signaling. Reactive oxygen species (ROS) accumulation increased markedly, accompanied by altered oxygen consumption rates—reduced in Swan71 cells and elevated in BeWO cells. Functionally, semaglutide suppressed Swan71 invasion through Matrigel by approximately three-fold. These findings suggest that semaglutide induces oxidative and metabolic stress in trophoblasts and is associated with altered mTOR-mediated signaling and reduced invasive potential. Such cellular alterations may contribute to compromised placental development and uterine vascular remodeling if exposure occurs near conception. While clinical data remain limited, this study provides mechanistic insight supporting caution in the use of semaglutide during the periconception period and underscores the need for targeted reproductive safety studies. Full article

Selenium (Se) is an important micronutrient that is required in very small amounts and plays a significant role in enhancing plant growth, stress resistance, and fruit quality. In this study, we investigated the effects of different sodium selenite concentrations (CK, 0 mg/L; Se1, 0.50 mg/L; Se2, 1.00 mg/L, Se3, 2.00; and Se4, 3.00 mg/L) on the growth, nutrient absorption, antioxidant capacity, and leaf metabolome of blueberry (Vaccinium corymbosum L. ‘Brigitta’) in hydroponic culture. Our results showed that moderate Se concentration (1.00 mg/L, Se2) had noticeable enhancements in key traits like taller plants, thicker stems, a greater number of leaves, and stem fresh weight, with increases of 60.23%, 61.90%, 36.05%, and 87.97%, respectively, compared to the CK. In addition, the appropriate application of Se fertilizer (1.0 mg/L, Se2) can enhance the absorption of macronutrients by plants, with the total contents of nitrogen (N), phosphorus (P), and potassium (K) increasing by 48.11%, 15.85%, and 14.25%, respectively, compared to CK. In comparison to CK, the content and accumulation of total Se rose dramatically under the Se4 treatment, showing increases of 2300% and 2514%. The contents of chlorophyll and antioxidant enzyme activities were maximized at Se2, while excessive Se (Se4) led to oxidative damage, as indicated by elevated MDA, H₂O₂, and O₂⁻ levels. Moreover, metabolomic analysis revealed that moderate Se concentration (Se2) significantly altered metabolic pathways related to aminoacyl-tRNA biosynthesis, arachidonic acid metabolism, and ABC transporters, with downregulation of key metabolites in sugar and organic acid metabolism (e.g., α-D-glucose-6-phosphate, L-lactic acid, maleic acid). In contrast, high Se concentration (Se4) disrupted these pathways and promoted volatile compound accumulation. These findings demonstrate that moderate Se application enhances blueberry growth and quality by regulating nutrient uptake, antioxidant defense, and primary metabolism, whereas excessive Se induces metabolic imbalance and oxidative stress. Overall, moderate Se fertilizer (1.00 mg/L) can significantly enhance the growth and quality of blueberries, while excessive selenium may have adverse effects. Full article

Pancreatic ductal adenocarcinoma (PDAC) is the most aggressive malignancy, characterized by rapid progression, early metastasis, and resistance to conventional therapies. Increasing evidence indicates that the behavior of residual tumor cells is strongly influenced by physicochemical properties of their microenvironment. Surface engineering strategies using nanostructured materials may therefore represent a complementary approach to modulating cancer cell activity. In this study, we investigated whether a graphene oxide (GO) aerosol nanofilm modifies the biological behavior of PDAC cells in vitro. The GO aerosol (4.5 g/L) was characterized using STEM, DLS, zeta potential measurements, LIBS, EDX, and FTIR spectroscopy. Ultrastructural analysis revealed thin, wrinkled GO sheets forming partially overlapping lamellar structures, while physicochemical characterization confirmed a highly oxidized stable nanomaterial. Human PDAC cell lines (BxPC-3 and AsPC-1) were cultured on GO-modified substrates to assess morphology (SEM), metabolic activity (XTT assay), migratory capacity (wound healing assay over 72 h), and expression of genes related to proliferation and epithelial–mesenchymal transition (EMT) by RT-qPCR. GO nanofilm significantly reduced cell viability and inhibited migration in both cell lines. SEM analysis demonstrated shortened cytoplasmic projections and altered membrane integrity. Gene expression profiling revealed cell line-dependent transcriptional responses, including modulation of components of the PI3K/AKT/mTOR pathway and EMT-associated markers. Collectively, our findings demonstrate that GO aerosol nanofilm alters PDAC cell morphology, viability, and migratory behavior in vitro. Surface-mediated modulation of tumor cell activity may represent a promising adjunct strategy for limiting residual cancer cell survival and metastatic potential. Full article

The twist angle serves as a geometric tuning parameter in two-dimensional layered materials, enabling modulation of interlayer coupling and band structures without altering the chemical composition. In this work, six commensurate twisted bilayer WSe₂ configurations with rotation angles of 0°, 9.4°, 13.14°, 21.9°, 27.8°, and 60° were systematically investigated using first-principles density functional theory. Structural optimization, together with calculations of electronic structures, density of states, charge redistribution, effective masses, and optical properties, was performed. The results show that AA (0°) and 2H (60°) stackings exhibit the largest and smallest interlayer separations, respectively, whereas intermediate twist angles yield similar average spacings but distinct local stacking registries. All configurations remain indirect-gap semiconductors, with the valence band maximum located at K and the conduction band minimum near the Q point along the K–Γ path. The band gap increases from 1.450 eV at 0° to 1.579 eV at 27.8°, before decreasing to 1.333 eV at 60°, indicating strong twist-angle modulation of interlayer coupling. Density-of-states analysis shows that the valence-band edge mainly originates from Se-p and W-d hybridized states, whereas the conduction-band edge is dominated by W-d states, with intermediate angles exhibiting enhanced band folding and localization features. Charge-density analyses further reveal notable interfacial charge redistribution, which is most pronounced at 9.4°. Optical responses in the in-plane directions are nearly identical and significantly stronger than those along the out-of-plane direction. Optical absorption mainly occurs in the ultraviolet region, with band-edge features appearing in the near-infrared range. Intermediate twist angles exhibit broader dielectric responses in the visible region and extended long-wavelength tails, indicating enhanced interband transition channels. These results demonstrate that twist-angle engineering enables effective tuning of electronic and optical properties in bilayer WSe₂, providing theoretical guidance for the design of tunable optoelectronic devices. Full article

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal oncological diseases, with a 5-year survival rate of approximately 13%—among the lowest in oncology. Poor survival is driven by aggressive tumor progression and metastasis, which may be influenced by the tumor microbiome. This study aimed to evaluate the role of microbiome in PDAC progression and metastasis. First, we assessed the microbial composition of control samples (surface swabs, empty paraffin, extraction controls, and sequencing controls) and removed contaminant taxa. Overall bacterial biomass was extremely low, with no significant differences in alpha or beta-diversity between tumor and normal tissue. Kocuria rosea was significantly enriched in tumors compared to normal tissue, and this difference persisted after decontamination. Metastatic tumors showed altered abundance of K. rosea and Herbaspirillum huttiense, whereas non-metastatic tumors differed in Lysobacter bugurensis, Caulobacter ginsengisoli, and H. huttiense relative to normal tissue. No global compositional differences were observed between KRAS-mutant and wild-type tumors; however, KRAS-mutant tumors exhibited differential enrichment of K. rosea and L. bugurensis relative to adjacent normal tissue. The PDAC microbiome harbors very low bacterial biomass and does not robustly distinguish tumor from normal tissue at the community level. Nonetheless, K. rosea emerges as a candidate taxon differentially enriched in PDAC, with potential stage- and KRAS-associated patterns. These findings highlight the need for orthogonal validation (qPCR, FISH, culture) and larger prospective cohorts to differentiate true biological associations from residual contamination or stochastic noise in low-biomass settings. Full article

Background and Aims: Understanding regulatory interactions between hepatitis B virus (HBV) and host factors is essential for the development of next generation host-directed antiviral therapies and the achievement of a functional HBV cure. Here, we investigated HBV-induced alterations in host gene expression in primary human hepatocytes (PHH) to identify host factors exploited by the virus for replication and persistence. Whole-transcriptome sequencing (WTS) of HBV-infected PHH identified host pathways with potential roles in the HBV life cycle. RNA interference-based functional screening of dysregulated candidate genes identified cyclin L1 (CCNL1) as a key host factor. RNAi-mediated knockdown of CCNL1 reduced HBV gene expression, including hepatitis B surface antigen (HBsAg). Mechanistically, CCNL1 regulates phosphorylation of the C-terminal domain (CTD) of RNA polymerase II (RNAPII) at serine 2 (S2), consistent with a role in transcriptional regulation. CCNL1 knockdown further reduced the binding of total and phospho- (Ser2/Ser5) RNAPII, pan-acetylated histone H3 (H3ac), and H3K27ac to HBV covalently closed circular DNA (cccDNA), indicating impaired cccDNA-dependent transcription. In addition, CCNL1 expression was elevated in chronic hepatitis B patients compared with those with resolved infection. Collectively, these data demonstrate that CCNL1 promotes HBV transcription and replication through modulation of RNAPII phosphorylation and chromatin-associated transcriptional activity, identifying CCNL1 as a potential host susceptibility factor for HBV. Importance: Hepatitis B virus infection remains a major threat to human health in areas with high prevalence. There is need to fully understand the complex interactions between the virus and human host factors/processes to support ongoing efforts to develop anti-HBV therapies that can be used with existing therapies to achieve a better cure. HBV relies on host cellular factors and biological processes to establish and maintain efficient infection, making host–virus interactions attractive targets for therapeutic intervention. Thus, identifying host factors that support and/or restrict HBV infection is essential for understanding the molecular basis of chronic HBV infection and for developing host-targeting anti-HBV drugs. This study identifies cyclin L1 (CCNL1) as a host susceptibility factor that promotes HBV transcription and replication through regulation of RNA polymerase II activity and or post-transcriptional mechanisms. Full article

Metabolic dysfunction, a hallmark of diet-induced obesity (DIO), is increasingly attributed to alterations in intestinal nutrient and electrolyte transport. Yet the mechanisms that drive obesity-associated functional alterations of intestinal transporters remain incompletely understood. In this context, the effects of a high-fat diet (HFD) induced obesity on sodium-dependent glucose co-transporter 1 (SGLT1), Na⁺/H⁺ exchanger 3 (NHE3), and Cl⁻/HCO₃⁻ exchangers (DRA/PAT1), the primary glucose, sodium, and chloride absorptive pathways in mice small intestinal villus cells, were investigated. SGLT1 activity significantly increased in intact villus cells and brush border membrane vesicles (BBMV) from HFD-fed mice. Kinetic analysis demonstrated reduced Km without a change in Vmax, indicating enhanced transporter affinity. Notably, SGLT1 mRNA and protein expression, including BBM localization, were unchanged. Basolateral Na⁺/K⁺-ATPase activity was decreased, excluding enhanced Na⁺ gradient generation as the mechanism for SGLT1 stimulation. In contrast, DRA/PAT1 activity was significantly increased in HFD-fed mice, and kinetic studies revealed elevated Vmax without a change in Km, indicating increased transport capacity. DRA/PAT1 mRNA, total protein, and BBM expression were all significantly elevated. NHE3 activity and expression remained unchanged. These findings demonstrate that DIO enhances intestinal glucose absorption by increasing SGLT1 affinity and chloride absorption by upregulating DRA/PAT1 transcription. These transporter-specific alterations may amplify nutrient absorption and contribute to metabolic dysregulation in obesity. Full article

Tea (Camellia sinensis L.) flowers have recently gained attention due to their bioactive composition, similar to that of tea leaves. They are used in food and cosmetic applications and show potential for medicinal use. However, catechins in tea flowers are highly susceptible to oxidation by polyphenol oxidase (PPO), leading to enzymatic browning. This process alters the phenolic profile and results in losses in appearance, nutritional value, and overall product quality. In this study, PPO from tea flowers was purified using affinity chromatography with a yield of 11.31% and a 91.90-fold purification. The molecular weight was determined to be approximately 42.67 kDa by SDS–PAGE. Substrate specificity studies revealed the highest activity toward catechin. Optimum pH and temperature were determined to be 5.0 and 40 °C, respectively. K_m and V_max values for catechin were 0.42 mM and 8333.3 EU·mL⁻¹·min⁻¹, respectively. The enzyme showed high stability at pH 5.0–7.0 and remained active for 60 min at 30 °C and 40 °C. L-cysteine was found to be the most effective of the inhibitors studied. These findings contribute to the understanding of the enzymatic browning mechanism of tea flower PPO and provide important data for enzyme control in food, cosmetic, and medical applications. Full article

Four perovskite manganite samples, La_0.80Ag_0.20MnO₃ (LA), La_0.78Eu_0.02Ag_0.20MnO₃ (LEA), La_0.80Pb_0.05Ag_0.15MnO₃ (LPA), and La_0.77Eu_0.03Pb_0.05Ag_0.15MnO₃ (LEPA), were prepared by the Pechini sol–gel method. The samples were characterized by X-ray diffraction, scanning electron microscopy, energy-dispersive spectroscopy, X-ray photoelectron spectroscopy, and a magnetic property measurement system. A systematic investigation was conducted into the individual effects of Eu and Pb doping, as well as their co-doping, on the microstructural, magnetic and magnetocaloric properties of the materials. The results show that all samples are mainly composed of a rhombohedral perovskite phase with the $\mathrm{R}\overline{3}\mathrm{c}$ space group, accompanied by a trace amount of Ag. Addition of Eu³⁺ and Pb²⁺ induces lattice contraction and expansion, respectively. Under the same processing conditions, the average crystallite and particle sizes of the LEA sample (45.3 nm and 0.18 μm) are smaller than those of the other three samples (69.6~80.6 nm and 0.38~0.44 μm), indicating that the introduction of Eu alone suppresses crystallization ability, which can be avoided through Eu/Pb co-doping. All samples undergo a second-order ferromagnetic–paramagnetic transition, and the Curie temperature T_C shifts to either lower or higher temperatures upon the introduction of Eu or Pb alone (from 310.8 K to 298.0 K or 318.0 K, respectively), which is attributed to the variation of the Mn³⁺/Mn⁴⁺ double-exchange (DE) interaction resulting from the ionic size mismatch and lattice distortion. In the LPA sample, an additional contribution arises from the altered Mn³⁺/Mn⁴⁺ ratio and enhanced DE interaction caused by the substitution of Pb²⁺ for Ag⁺. By modifying the Eu/Pb ratio, the T_C of the LEPA sample was tuned to 299.3 K, and its maximum magnetic entropy change was enhanced to 3.90 J·kg⁻¹·K⁻¹ ($∆$H = 2 T). These results indicate that multicomponent synergistic regulation can improve the magnetocaloric performance of La-based perovskite manganites, providing a useful strategy for the development of room-temperature magnetic refrigeration materials. Full article

Clear cell renal cell carcinoma (ccRCC) is characterized by marked biological heterogeneity, which limits the prognostic accuracy of conventional clinicopathological models. Increasing attention has therefore focused on identification of biomarkers that can enhance risk stratification throughout all stages of the disease. Starting from the current state of the art, this narrative review summarizes and critically appraises the evidence published over the past decade regarding prognostic biomarkers in ccRCC. The analysis is structured into four overarching domains: (i) genomic biomarkers, covering somatic alterations and transcriptomic signatures; (ii) tissue-based biomarkers, including immunohistochemical surrogates and immune microenvironment features; (iii) circulating biomarkers, such as systemic inflammation parameters and indices; and (iv) integrated predictive models, represented by emerging multi-omic approaches. Going through the broad framework of potential prognostic biomarkers, emphasis is placed on their individual and integrative value in relation to classic clinical-pathological factors and survival parameters. At the tissue level, chromosome 3p-related alterations constitute a central molecular feature of ccRCC. Among these, BAP1 loss has emerged as one of the most consistently validated indicators of aggressive tumor behavior. Disruption of the SETD2/H3K36me3 axis and immune-related biomarkers, including PD-L1 expression, have demonstrated prognostic associations in selected settings, although with variable and context-dependent performance. In the circulating compartment, plasma KIM-1 has shown prognostic relevance following nephrectomy, while postoperative detection of circulating tumor DNA (ctDNA) may identify patients at increased risk of recurrence. However, limited analytical sensitivity and methodological heterogeneity currently restrict the broader clinical applicability of ctDNA-based strategies. Systemic inflammatory indices, such as the neutrophil-to-lymphocyte ratio, show reproducible associations with outcomes but largely reflect host inflammatory status rather than tumor-specific biology. However, no single biomarker currently supports routine prognostic implementation in ccRCC. Future progress will likely depend on integrative models combining genomic, tissue-based, immune, and circulating parameters with established clinical variables. Prospective validation and clear demonstration of incremental clinical utility will be essential before such strategies can meaningfully inform therapeutic decision-making. Full article

Maternal infection (MI) can increase the risk of neurodevelopmental and behavioural changes. This study examined MI-induced changes in motor coordination through the inflammatory-pathway-mediated epigenetic status of Reln. On gestational day (GD) 10, rats were assigned as (i) Control (Ctrl); (ii) Cronobacter sakazakii (CS) infection on GD-10 through recto-vaginal colonization; (iii) Negative Control (NC) [infected with C. sakazakii and treated with dimethyl sulfoxide (DMSO) 1 h before and 24 h after infection]; and (iv) C. sakazakii-infected rats treated with matrix metalloproteinase inhibitor (MMPI), 1 h before and 24 h after infection (CS + MMPI). Offspring were subjected to footprint analysis and the ladder rung walking test, which revealed that MI caused significant deficits in motor coordination. In addition, MI activated complement components—a disintegrin and metalloproteinase with thrombospondin motifs-1 (ADAMTS-1, C5a)—as well as proinflammatory cytokines such as interleukin-6 (IL-6) and matrix metalloproteinases (MMP-2, MMP-3, and MMP-9). Furthermore, the levels of DNA methyltransferase 3 alpha (DNMT3A), methyl-CpG-binding protein 2 (MeCP2), and histone H3 lysine 4 trimethylation (H3K4me3) were elevated in the CS and NC groups. Concurrently, the level of Reln promoter methylation increased; as a result, mRNA and protein, as well as postsynaptic density protein-95 (PSD-95), levels were decreased. Overall, the findings suggest that MI altered MMP-2/3/9 activity, H3K4me3, and the methylation of Reln, thereby affecting reelin, synaptic protein expression, and motor coordination in an experimental meningitis rat model. Full article

Diffuse midline gliomas (DMGs) are rare but highly aggressive central nervous system (CNS) tumors that can present in both pediatric and adult populations. These tumors were redefined in the 2016 WHO classification of CNS tumors based on integrated histopathological and molecular features, and were initially designated as “DMG, H3 K27M-mutant”. In the 2021 WHO update, DMGs were incorporated into the newly defined category of primarily pediatric-type diffuse high-grade gliomas, and nomenclature was changed to “DMG, H3 K27-altered” to encompass additional molecular drivers beyond the canonical H3 K27M mutation. Clinically, DMGs arise as expansile, infiltrating tumors within midline structures and may present as non-enhancing or enhancing lesions on imaging. Diagnosis is based on neuroimaging and molecular confirmation by immunohistochemistry or sequencing when tissue is available. DMGs are categorized as WHO grade 4 malignant tumors due to their aggressive biology leading to rapid and infiltrative growth. Owing to their deep and midline location, surgical resection is typically not feasible. Radiation therapy is the backbone of treatment, but there is no standard regimen of chemotherapy that has demonstrated durable efficacy. Recent progress in therapeutic approaches has led to a major breakthrough on 6 August 2025 when the U.S. Food and Drug Administration granted the accelerated approval of dordaviprone (ONC201), marking it as the first systemic therapy for progressive DMG harboring H3 K27M mutation. Other novel approaches, including chimeric antigen receptor (CAR) T-cell directed therapies and convection-enhanced delivery, are actively under investigation. We aim to comprehensively review DMGs, including the recent insights into their biology, the evolving therapeutic landscape, and the opportunities to fuel this new momentum against one of the most formidable gliomas. Full article

This study systematically compared the induction and resuscitation characteristics of the viable but non-culturable (VBNC) state in Klebsiella pneumoniae FY170-1 following sublethal exposure to sodium hypochlorite (NaClO) or glutaraldehyde (GA). Treatment with 30 mg/L NaClO or 60 mg/L GA for 60 min reduced culturability to below the detection limit (<1 CFU/mL). However, CTC staining showed that 50.80% and 63.44% of cells, respectively, retained respiratory activity, while SYTO 9/PI staining indicated that membrane integrity was largely preserved, consistent with induction of the VBNC state. Scanning electron microscopy revealed distinct morphological alterations in the two groups. NaClO-induced VBNC cells showed surface depressions and wrinkling, consistent with oxidative damage, whereas GA-induced cells exhibited filamentous and net-like surface structures, consistent with aldehyde-mediated cross-linking. Among the tested additives, sodium succinate showed the strongest resuscitation-promoting effect under the experimental conditions, with OD600 increasing after approximately 2 h of incubation. Post-resuscitation analysis further revealed marked differences between the two VBNC states. In resuscitated NaClO-induced VBNC cells, ATP partially recovered, but reactive oxygen species remained elevated and catalase activity showed little recovery. In contrast, resuscitated GA-induced VBNC cells exhibited lower ATP recovery but more rapid normalization of ROS and better recovery of oxidative stress-related parameters. Total protein analysis and SDS-PAGE further supported distinct patterns of protein-level alteration between the two treatments. Overall, these findings suggest that NaClO and GA induce phenotypically distinct VBNC states in K. pneumoniae, with different recovery behaviors and stress response profiles. Sodium succinate was identified as the most effective recovery-promoting additive under the tested conditions. These results highlight the risk of underestimating bacterial survival when culturability is used as the sole indicator of disinfection efficacy and support the need for more comprehensive viability assessment. Full article