Search Results (9,339)

Cellular senescence can occur with similar phenotypes in normal cells, during aging, and in tumor cells, spontaneously or after cytostasis. The fall or increase in proliferative activity are key aspects of the respective conditions, in which the levels of reactive oxygen species can vary, affecting the cellular redox homeostasis. This work aimed to study the relationships between senescence and transformation by comparing cells with different proliferative activities and phenotypes attributable to transformation (NIHs cultures) or senescence (NIHv cultures), before and after incubation with hydrogen peroxide. Both cultures were derived from the NIH/3T3 cell line, which was used here as a reference (NIHb), after the serum starvation. Our experimental model can be representative of the heterogeneity of cell subpopulations, with different degrees of transformation and senescence, found in some tumors. The characterization of the functional properties of NIHb, NIHs, and NIHv cells was performed by a morphocytometric analysis of the cell cycle progression, mitochondrial and lysosomal content/activity, and superoxide anion production. The efficiency of the lysosomal compartment was also assessed by estimating the autophagic activity and measuring lipofuscin autofluorescence. Comparisons of nuclear and cytoplasmic parameters before and after the incubation with hydrogen peroxide revealed differences in the expression and modulation of cellular senescence patterns. The treatment effects were very limited in the NIHb culture; the senescence condition was essentially maintained in the NIHv cells, while the most relevant changes were found in the NIHs cells. In the latter, the acquisition of the senescent phenotype, also demonstrated by the positivity of SA-β-galactosidase, was correlated with a decrease in proliferative activity and a change in the content/activity of the mitochondria and lysosomes, which showed similarities with the basal senescence conditions of NIHv cells. In NIHs cells, increased autophagy events and lipofuscin accumulation also indicate the establishment of cytoplasmic dynamics typical of senescence. The variable responses to hydrogen peroxide, besides depending on the different basal cytokinetic activity of the cultures examined, appeared to be related to the specific cell redox state resulting from the balance between endogenous ROS and those produced after treatment. Especially in NIHs cells, the slowing down of the cell cycle was linked to dynamic interconnections between the mitochondrial and lysosomal compartments. This would indicate that transformed cells, such as NIHs, may express morpho-functional aspects and markers typical of cellular senescence, as a consequence of the modulation of their redox state. Full article

The evolving legal landscape has increased marijuana accessibility across the United States, including for medical use to manage clinical symptoms among people with HIV. The effects of marijuana use remain understudied in youth with HIV (YWH), who face lifelong antiretroviral therapy (ART) and an elevated risk of developing comorbidities. This study applied a multi-modal approach, including plasma biomarker analysis, peripheral blood cell phenotyping, and transcriptome profiling, to examine the effects of recreational marijuana alone, tobacco alone, or marijuana combined with tobacco in virally suppressed YWH (≤50 RNA copies/mL) on ART compared to youth without HIV and YWH who used no substance. Marijuana use alone was associated with elevated IL-10 levels and normalization of pro-inflammatory genes and pathways, suggesting an immunomodulatory effect. Conversely, tobacco use alone or combined with marijuana was linked to increased IL-1β levels and heightened pro-inflammatory responses, including upregulation of genes involved in inflammasome activation. This study is the first to demonstrate GPR15 upregulation and potential marijuana-associated epigenetic modulation in HIV-suppressed youth. The findings identify potential markers for early detection of inflammation-related comorbidities in YWH, particularly among those exposed to tobacco and underscore the need for targeted profiling to guide personalized monitoring and early substance use intervention strategies for YWH. Full article

Chronic obstructive pulmonary disease (COPD) is a heterogeneous syndrome with multiple clinical and inflammatory phenotypes. The coexistence of bronchiectasis, known as bronchiectasis–COPD overlap (BCO), identifies a subgroup with increased morbidity and mortality. Non-invasive breath analysis using electronic noses (e-noses) has shown promise in identifying disease-specific volatile organic compound (VOC) patterns (“breathprints”). Our aim was to evaluate the ability of an e-nose to differentiate between COPD and BCO patients, and to assess its utility in detecting inflammatory endotypes (neutrophilic vs. eosinophilic). In a monocentric, prospective, real-life study, 98 patients were enrolled over nine months. Forty-two patients had radiologically confirmed BCO, while fifty-six had COPD without bronchiectasis. Exhaled breath samples were analyzed using the Cyranose 320 e-nose. Principal component analysis (PCA) and discriminant analysis were used to identify group-specific breathprints and inflammatory profiles. PCA revealed significant breathprint differences between BCO and COPD (p = 0.021). Discriminant analysis yielded an overall accuracy of 69.6% (AUC 0.768, p = 0.037). The highest classification performance (76.8%) was achieved when distinguishing eosinophilic COPD from neutrophilic BCO. These findings suggest distinct inflammatory profiles that may be captured non-invasively. E-nose technology holds potential for the non-invasive endotyping of COPD, especially in identifying neutrophilic BCO as a unique inflammatory entity. Breathomics may support early, personalized treatment strategies. Full article

Background/Objectives: Fatigue is a highly prevalent and burdensome symptom among individuals with Marfan syndrome (MFS), yet its heterogeneity and underlying psychosocial and clinical correlates remain underexplored. This study aimed to identify and characterize distinct fatigue-related profiles in MFS patients using a data-driven cluster analysis approach. Methods: A cross-sectional study was conducted involving 127 patients with MFS from a specialized connective tissue disorder center in Italy. Participants completed self-reported measures of fatigue severity (Fatigue Severity Scale, FSS), depressive symptoms (Patient Health Questionnaire-9, PHQ-9), and insomnia (Insomnia Severity Index, ISI). The body mass index (BMI) and clinical data were also collected. A t-distributed stochastic neighbor embedding (t-SNE) analysis was performed to reduce dimensionality, followed by hierarchical clustering (Ward’s method), exploring solutions from k = 2 to k = 10. The optimal cluster solution was identified based on silhouette scores and clinical interpretability. Results: Three distinct clusters emerged: (1) a cluster characterized by low fatigue with minimal psychological and sleep-related symptoms (younger patients, lower PHQ-9 and ISI scores), (2) a cluster characterized by moderate fatigue with moderate psychological and sleep-related symptoms (intermediate age, moderate PHQ-9 and ISI scores), and (3) a cluster characterized by high fatigue with elevated psychological and sleep-related symptoms (older patients, higher PHQ-9, ISI, and FSS scores). Significant differences were observed across clusters in age, BMI, depressive symptoms, insomnia severity, and fatigue levels (all p < 0.05). Conclusions: Our findings highlight the heterogeneity of fatigue experiences in MFS and suggest the importance of profiling patients to guide personalized interventions. This approach may inform precision medicine strategies and enhance the quality of life for individuals with this rare disease. Full article

The immune response to SARS-CoV-2 infection involves significant alterations in the phenotype and function of natural killer (NK) cells. This study aimed to investigate the dynamic changes in NK cell subsets during COVID-19 by analyzing their activation and inhibitory markers [CD3, CD14, CD16, CD19, CD25, CD45, CD56, CD57, CD69, CD159a (NKG2A), CD159c (NKG2C), CD314 (NKG2D), CD335 (NKp46)], cytotoxic potential (perforin, interferon-gamma, granzyme B), and direct cytotoxicity against a newly genetically modified K562 cell line. Peripheral blood samples were collected from COVID-19 patients on days 3–5 and day 30 post-symptom onset and were compared to healthy controls. 16-color flow cytometry analysis revealed distinct shifts in NK cell subpopulations, characterized by increased expression of the inhibitory receptor NKG2A and the activating receptors NKG2D and NKG2C, particularly in the CD56⁺CD16⁻ subset. Elevated IFN-γ production on day 30 suggested a recovery-phase immune response, while the persistent upregulation of NKG2A indicated an ongoing regulatory mechanism. The CD16⁺CD56⁻ subpopulation exhibited increased expression of the markers CD69 and CD25 over time; however, its cytotoxic potential, assessed through granzyme B levels and direct cytotoxicity assays, remained lower than that of healthy controls. Significant correlations were observed between CD57 and CD69 expression, as well as NKp46 and IFN-γ production, highlighting a coordinated balance between activation and regulatory mechanisms. These findings suggest that NK cells undergo functional adaptation during COVID-19, displaying signs of partial exhaustion while retaining antiviral potential. Understanding the interplay between NK cell activation and suppression may provide valuable insights into immune dysregulation in COVID-19 and inform potential therapeutic interventions. Full article

The Tumour Microenvironment (TME) is pivotal for melanoma progression and contributes to therapy resistance. While dermal cell involvement is well established, the role of epidermal cells remains less defined. To explore the contribution of Normal Human Keratinocytes (NHKs) to melanoma biology, we investigated the modification of gene and protein expression of NHKs exposed to melanoma-conditioned medium or maintained in a co-culture system. The analysis focused on pathways related to proliferation, inflammation, Extracellular Matrix (ECM) remodelling, and cell adhesion. Due to the well-documented melanoma–fibroblast crosstalk, Normal Human Fibroblasts (NHFs) and Cancer-Associated Fibroblasts (CAFs) were used as comparative references. Keratinocyte gene expression changes under the influence of melanoma secretome only partially overlapped with those of NHFs and CAFs, indicating cell-type-specific responses. Exposure to melanoma-conditioned medium induced the upregulation of bFGF, CXCL-16, TIMP-2, and E-cadherin in NHKs, alongside downregulating TGF-β and MMP-9. Although bFGF is a recognized pro-tumorigenic factor, the modulation of CXCL-16, TIMP-2, and TGF-β may reflect a protective response. Notably, under co-culture conditions, NHKs exhibited a pronounced pro-inflammatory and ECM-remodelling phenotype, characterized by elevated production of cytokines (IL-1α, IL-1β, and IL-8) and ECM-degrading enzymes (MMP-7, 9, 12, and 13), indicative of a pro-tumoral feature. Collectively, these findings underscore an active role for NHKs in melanoma initiation and progression. Full article

Proinflammatory cytokines such as IL-1β, IL-6, and TNF-α are key mediators of inflammatory bone loss and are commonly described as inhibitors of osteoblast function. However, their effects on osteogenesis remain controversial, likely due to the differences in the cell models and experimental settings in in vitro studies. We recently showed that these cytokines significantly enhanced the mineralization of primary human osteoblast-like cells (OBs). Here, we provide the first analysis of cytokine effects on the osteogenesis of the widely used human osteoblastic cell line hFOB 1.19 and compare them to primary OBs. Unexpectedly, all three cytokines significantly inhibited mineralization in hFOB 1.19 cells without affecting the proliferation. IL-1β and TNF-α also suppressed ALP activity, whereas IL-6 acted ALP-independent but increased the osteogenic marker expression despite the reduced mineralization, indicating a possible uncoupled differentiation and mineralization. Morphological and transcriptional analyses indicated that hFOB 1.19 cells represent an earlier osteogenic differentiation stage, while primary OBs show phenotypic heterogeneity and donor-dependent expression profiles. These data demonstrate that proinflammatory cytokines can have severely different effects on the osteogenesis of different cell models, supported by the highly contradictory findings reported in the literature. Nevertheless, elucidating the mechanisms underlying the inhibition of osteogenesis in hFOB 1.19 cells may provide important insights into the cell model and differentiation-stage-specific cytokine effects. Full article

Temperate forest ecosystems in Korea are currently undergoing a successional transition from Pinus densiflora Siebold & Zucc. (evergreen conifer) communities to Quercus mongolica Fisch. ex Ledeb. (deciduous broadleaf) communities. This study aimed to assess interspecific differences in ecological responses to climate change [Representative Concentration Pathway (RCP) 4.5] by evaluating changes in ecological niche characteristics and species distribution. Controlled-environment experiments, principal component analysis (PCA), and MaxEnt species distribution modeling were employed to quantify and predict ecological shifts in the two dominant species under climate change scenarios. Both species exhibited increases in niche breadth and interspecific overlap under climate change conditions. However, Q. mongolica showed a more pronounced increase in niche breadth compared to P. densiflora, indicating greater ecological flexibility and adaptive potential to warming conditions. According to the MaxEnt model projections, climate change is expected to result in an approximate 30% reduction in suitable habitat for P. densiflora in lowland areas. In contrast, Q. mongolica is projected to expand its suitable habitat by over 80%, notably in both low-elevation (below 800 m) and high-elevation (above 1400 m) zones, without being restricted to any specific altitudinal range. Our findings suggest that climate change may increase ecological similarity between P. densiflora and Q. mongolica, thereby raising the potential for interspecific competition. This convergence in niche traits could contribute to an accelerated successional transition, although actual competitive interactions in natural ecosystems require further empirical validation. Consequently, Korean forests are likely to transform into predominantly deciduous forest ecosystems under future climate conditions. Full article

Spectral reflectance of plants can be readily associated with physiological and biochemical parameters. Thus, relating spectral data to amino acid contents in different genetic materials provides an innovative and efficient approach for understanding and managing genetic diversity. Therefore, this study had two objectives: (I) to differentiate genetic materials according to amino acid contents and spectral reflectance; (II) to establish the relationship between amino acids and spectral bands derived from hyperspectral data. The research was conducted with 32 soybean genetic materials grown in the field during the 2023–2024 crop year. The experimental design involved randomized blocks with four replicates. Leaf spectral data were collected 60 days after plant emergence, when the plants were in full bloom. Three leaf samples were collected from the third fully developed trifoliate leaf, counted from top to bottom, from each plot. The samples were taken to the laboratory, where reflectance readings were obtained using a spectroradiometer, which can measure the 350–2500 nm spectrum. Wavelengths were grouped as means of representative intervals and then organized into 28 bands. Subsequently, the leaf samples from each plot were subjected to quantification analyses for 17 amino acids. Then, the soybean genotypes were subjected to a PCA–K-means analysis to separate the genotypes according to their amino acid content and spectral behavior. A correlation network was constructed to investigate the relationships between the spectral variables and between the amino acids within each group. The groups formed by the different genetic materials exhibited distinct profiles in both amino acid composition and spectral behavior. Leaf reflectance data proved to be efficient in identifying differences between soybean genotypes regarding the amino acid content in the leaves. Leaf reflectance was effective in distinguishing soybean genotypes according to leaf amino acid content. Specific and high-magnitude associations were found between spectral bands and amino acids. Our findings reveal that spectral reflectance can serve as a reliable, non-destructive indicator of amino acid composition in soybean leaves, supporting advanced phenotyping and selection in breeding programs. Full article

Aeromonas is a genus of Gram-negative, facultatively anaerobic bacteria commonly found in aquatic environments and increasingly recognized as opportunistic pathogens. Among them, Aeromonas dhakensis stands out for its high virulence and antimicrobial resistance, but it is often misidentified due to its phenotypic similarity with A. hydrophila and A. caviae. In this study, a microorganism was isolated from the peritoneal fluid of a patient with signs of intra-abdominal infection. MALDI-TOF MS initially suggested A. hydrophila or A. caviae, but the identification was confirmed with high confidence only after further molecular analyses and the use of genome-based tools, which identified the organism as A. dhakensis. This was further supported by phylogenomic and ANI analysis. Resistome analysis revealed intrinsic resistance genes, including a chromosomal class C β-lactamase and an imiH-type carbapenemase, consistent with the observed carbapenem resistance. No plasmid-mediated carbapenemases were found. These findings underscore the limitations of MALDI-TOF in identifying Aeromonas species in certain cases and highlight the value of genomic approaches for accurate species determination and resistance profiling, especially for isolates from sterile body fluids. Full article

Background: Left ventricular (LV) non-compaction (NC) is a rare ventricular phenotype characterized by a thin compacted epicardial layer and an extensive non-compacted endocardial layer with prominent LV trabeculations and deep intertrabecular recesses. According to the recent literature, no information is available regarding the abnormalities of the aortic valve annulus (AVA) in LVNC. Therefore, the aim of the present study was to perform a detailed analysis of the AVA by three-dimensional speckle-tracking echocardiography (3DSTE) in LVNC patients and to compare the findings with matched healthy controls. Methods: The present study comprised 21 isolated LVNC patients, from which 9 cases were excluded due to inferior image quality. The remaining group consisted of 12 patients with isolated LVNC (mean age: 54.6 ± 13.6 years, 7 males). Jenni’s echocardiography criteria served as a definition of LVNC. The 12 patients’ results were compared to 38 healthy age- and gender-matched controls (mean age: 48.2 ± 8.0 years, 19 males). Subgroups of patients having a greater end-diastolic versus end-systolic AVA area were also compared. Results: Most of AVA dimensions did not differ significantly between LVNC patients and controls; however, most LVNC patients showed a larger end-diastolic AVA area (9 out of 12, 75%), which was a significantly larger ratio as seen in matched controls (11 out of 38, 29%, p < 0.05). Aortic valve annular plane systolic excursion (AAPSE) proved to be significantly reduced in all LVNC patients (1.12 ± 0.24 cm vs. 0.78 ± 0.28 cm, p < 0.05) and in LVNC subjects with a greater end-diastolic AVA area (1.11 ± 0.21 cm vs. 0.72 ± 0.21 cm, p < 0.05). Basal LV radial (RS) and longitudinal (LS) strains were reduced in healthy adults with a greater end-diastolic AVA area as compared to cases with a greater end-systolic AVA area. In LVNC, not only basal LV-RS and LV-LS, but also LV circumferential strain (CS) proved to be reduced regardless of whether the AVA was greater in end-diastole or in end-systole. Conclusions: In patients with isolated LVNC, the AVA is not dilated; however, the presence of a greater end-diastolic AVA area is observed more frequently than in healthy controls. AAPSE and basal LV-RS, LV-LS and LV-CS values are significantly reduced in LVNC irrespective of whether the end-systolic or end-diastolic AVA area is greater. Full article

Background/Objectives: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is an urgent public health threat due to its rapid dissemination and resistance to last-line antibiotics. Cefiderocol (FDC), a novel siderophore cephalosporin, targets resistant Gram-negative pathogens by exploiting bacterial iron uptake mechanisms. However, resistance to FDC is emerging among Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae strains. This study characterizes a spontaneous FDC-resistant subpopulation (IHC216) derived from a KPC-producing strain (KPNMA216) using comprehensive genomic, transcriptional, and phenotypic analyses. Methods: Given the whole-genome sequencing results, where mutations were identified in genes involved in transcriptional regulation and membrane permeability (ompC) among others, in the present work we further explore their potential implications and conduct a more detailed analysis of the IHC216 genome. A qRT-PCR analysis highlighted significant downregulation of classical siderophore-mediated iron acquisition systems (fepA, cirA, iroN) and upregulation of alternative iron uptake pathways (iucA, fiU), reflecting a switch in iron acquisition strategies. Results: A notable downregulation of bla_KPC-163 correlated with restored susceptibility to carbapenems, indicating collateral susceptibility. Altered expressions of pbp2 and pbp3 implicated adaptive changes in cell wall synthesis, potentially affecting FDC resistance mechanisms. Furthermore, enhanced oxidative stress responses via upregulated sodC expression and increased capsule production were observed. Conclusions: These findings underscore the complex interplay of genetic and transcriptional adaptations underlying FDC resistance, highlighting potential therapeutic vulnerabilities. Full article

Background/Objectives: The progression of antibiotic resistance is increasingly recognized as a dynamic and time-dependent phenomenon, challenging conventional diagnostics that define resistance as a binary trait. Methods: Biomolecules have fingerprints in Fourier-transform infrared spectroscopy (FTIR). The targeting of specific molecular groups, combined with principal component analysis (PCA) and machine learning algorithms (ML), enables the identification of bacteria resistant to antibiotics. Results: In this work, we investigate how effective classification depends on the use of different numbers of principal components, spectral regions, and defined resistance thresholds. Additionally, we explore how the time-dependent behavior of certain spectral regions (different biomolecules) may demonstrate behaviors that, independently, do not capture a complete picture of resistance development. FTIR spectra were obtained from Staphylococcus aureus exposed to azithromycin, trimethoprim/sulfamethoxazole, and oxacillin at sequential time points during resistance induction. Combining spectral windows substantially improved model performance, with accuracy reaching up to 96%, depending on the antibiotic and number of components. Early resistance patterns were detected as soon as 24 h post-exposure, and the inclusion of all three biochemical windows outperformed single-window models. Each spectral region contributed distinctively, reflecting biochemical remodeling associated with specific resistance mechanisms. Conclusions: These results indicate that antibiotic resistance should be viewed as a temporally adaptive trajectory rather than a static state. FTIR-based biochemical profiling, when integrated with ML, enables projection of phenotypic transitions and supports real-time therapeutic decision-making. This strategy represents a shift toward adaptive antimicrobial management, with the potential to personalize interventions based on dynamic resistance monitoring through spectral biomarkers. Full article

Background: Transcription factors have been linked to changes in various physiological processes, such as attractive and rewarding phenotypes during plant–pollinator interactions. In the genus Petunia, most species are pollinated by bees, but hawkmoth- and bird pollination are also observed. Here, we aimed to test the hypothesis that species with the same pollination syndrome evolved through convergence, while differences in pollinators indicate divergence. We selected six genes (MYB-FL, DFR, EOBII, ODO1, BPBT, and NEC1) involved in establishing pollination syndromes to explore the potential role of cis-regulatory elements in shifts among pollination syndromes, attracting and rewarding pollinators. Methods: We retrieved the genomic sequences of genes from the genomes of four Petunia species, which exhibit distinct pollination syndromes. We analyzed the cis-regulatory elements, focusing on the structure and composition of motifs, and inferred the functions of these transcription factors using Gene Ontology analysis. Results: All sequences were highly conserved among species, with variations in promoter motif structure and TF binding sites. The evolutionary relationships among the genes closely reflected the species’ phylogeny. Likewise, regulatory elements and gene structure mostly followed the species’ evolutionary history. However, different pollination syndromes are present, and there is an unexpected lack of convergence between the two bee-pollinated species. Conclusions: Our findings showed that the most recent common ancestor of these species better predicts relationships among gene regulatory elements than does the pollination syndrome. To fully understand the evolution of pollination syndromes in Petunia, additional studies are needed to analyze entire pathways and compare genomes and transcriptomes. Full article

Studying comprehensive performance is fundamental for the effective utilisation of broomcorn millet (Panicum miliaceum L.) germplasm resources and breeding of new varieties. However, compared with other major crops, research on broomcorn millet germplasm resources is limited, and the trait variations of broomcorn millet are unclear. In this study, three qualitative and seven quantitative traits of 933 broomcorn millet core collections were analysed to provide the basis for improving utilisation of broomcorn millet germplasm resources. The seed colour was a strong phenotypic trait and had eight variants. The 933 resources exhibited three panicle types: lateral (74.5%), scattered (18.4%), and compact (7.1%). They exhibited two inflorescence colours: green (54.7%) and purple (45.3%). Pearson’s correlation analysis revealed that 1000-seed weight significantly correlated with plant height, length of panicle, and number of main stem segments. The period of duration positively correlated with 1000-seed weight but negatively correlated with the number of uniserial panicles. Cluster analysis based on the quantitative traits indicated that all resources were divided into three groups, and each group had its respective characteristics. The analysis of core germplasm resources of broomcorn millet in this study provided a basis to explore excellent genes and for breeding of excellent varieties. Full article