Search Results (115)

iPSC technology is well established in mammals but remains underdeveloped in non-mammalian species. A major barrier to generating avian iPSCs has been the lack of species-specific reprogramming factors and culture conditions capable of supporting self-renewal in avian pluripotent stem cells. Here, we report the generation of chicken iPSCs (ciPSCs) using a cocktail of seven chicken transcription factors (T7: Oct4, Sox2, Sox3, Klf4, c-Myc, Nanog, and Lin28B) combined with an optimized avian culture system. Transcriptomic and functional analyses identified Sox3, rather than Sox2, as the predominant SoxB1 factor in avian reprogramming. The resulting ciPSCs exhibited stable self-renewal for over 40 passages, expressed core pluripotency markers, differentiated into all three germ layers, and were transcriptionally similar to chicken ESCs. In chimera assays, ciPSCs contributed to somatic, extra-embryonic, and germline lineages, giving rise to gonadal PGC-like cells that did not acquire full germline competence. We further demonstrate that the T7 system generates iPSCs from quail, duck, peacock, zebra finch, and pigeon, and that duck iPSCs can form interspecies chimeras with donor cells detected in the host gonads. These findings establish a generalizable platform for avian iPSC generation with applications in developmental biology and germline preservation of endangered species. Full article

Background/Objectives: Hematopoietic stem cells (HSCs) sustain lifelong blood cell production and hold therapeutic promise, yet their ex vivo expansion remains constrained by an incomplete understanding of the metabolic and cellular mechanisms governing self-renewal. In this study, we investigated whether boron compounds boric acid (BA), sodium pentaborate pentahydrate (NaB), and sodium 2-pentaborate pentahydrate-8 (Na2B8) can promote the expansion of mouse HSCs by modulating key stem cell populations linked to metabolic fitness. Methods: Lineage-negative (Lin−) cells were magnetically isolated and treated with each boron compound for four days, followed by flow cytometric analysis of c-Kit, Sca-1, Lin-c-Kit+Sca-1+ (LSK), and LSKCD34^Low HSC-enriched subsets. Results: Our results show that boron derivatives exert distinct effects on these cellular markers. Notably, NaB treatment significantly increased the Lin-Sca-1+ cell fraction, while Na2B8 elevated both LSK and LSKCD34^Low ratios. Furthermore, the BA+NaB combination produced a statistically significant proliferative effect on Sca-1+ and c-Kit+ (CD117) cells. Conclusions: These findings indicate that specific boron compounds enhance ex vivo HSC expansion through yet-to-be-defined mechanisms that underpin HSC self-renewal. Further mechanistic studies are warranted to delineate the precise metabolic targets, but these results highlight boron compounds as promising tools for improving HSC expansion strategies. Full article

Listeria innocua is a bacterium frequently detected in food and food production plants (FPPs). Understanding the heterogeneity of L. innocua food isolates is essential for predicting potential food safety threats and developing preventive and control measures. This study aimed to characterize L. innocua isolated from raw drinking milk by investigating the genomic features related to virulence, antimicrobial resistance, and persistence using whole-genome sequencing (WGS), along with phenotypic antimicrobial susceptibility testing using the disk diffusion method. All ten isolates analyzed in this study belonged to sequence type (ST) 492 and were distantly related to the reference strain. A total of 80 virulence-associated genes were identified, including the complete Listeria Pathogenicity Islands-3 (LIPI-3) and LIPI-4 clusters typically found in virulent L. monocytogenes clones, as well as 66 additional genes involved in adhesion, invasion, motility, post-translational modification, regulation, immune modulation, and stress survival. Stress survival islet 2 (SSI-2) and genes encoding the Clp protease complex (clpC, clpE, clpP), which support both persistence and virulence, were also detected, whereas LIPI-1 and internalin genes were not detected. The antimicrobial resistance determinants included fosX, lin, norB, sul, and three multidrug efflux pumps (lde, mdrL and mdrM). Mobile genetic elements (plasmids, prophages, or transposons) were not detected. All isolates were phenotypically susceptible to benzylpenicillin, ampicillin, meropenem, erythromycin, and trimethoprim–sulfamethoxazole. These findings underscore the importance of ongoing genomic surveillance of L. innocua in food environments and highlight the need to assess the potential risk posed by specific lineages, such as ST492, to food safety. Full article

Background: Growing antibiotic resistance and the limited availability of key components in standard Helicobacter pylori treatments have driven the search for effective alternatives. Minocycline, with its broad-spectrum activity and favorable pharmacokinetics, has emerged as a promising substitute. This meta-analysis compares the safety and efficacy of minocycline-containing bismuth quadruple therapy (MBQT) to conventional first-line BQT regimens, incorporating data from the recent study by Lin et al. Methods: The inclusion criteria were randomized controlled trials (RCTs) with a target population of both treatment-naïve and previously treated patients diagnosed with Helicobacter pylori (H. pylori) infection. The intervention received by eligible patients was a minocycline–bismuth quadruple therapy (MBQT) regimen containing bismuth, minocycline, proton pump inhibitors (PPI), and any additional antibiotic with a minimum period of 2 weeks of administration. We excluded study designs other than RCT and clinical trials that include patients without confirmed H. pylori infection, animal populations, in vitro experiments, and reports of other outcomes that did not include a minimum intervention duration of 2 weeks. A comprehensive literature search was conducted on PubMed, EMBASE, Cochrane Library, and ScienceDirect from inception to 20 May 2025. After screening via Rayyan, data were extracted on an Excel spreadsheet. Quality was assessed using the Cochrane RoB 2.0 tool. Eligible randomized controlled trials (RCTs) were included and analyzed using RevMan 5.4. Outcomes assessed were intention-to-treat and per-protocol eradication rates. Adverse effects were compared among therapies. A random-effects model was used; an I² < 50% and p-value < 0.05 indicated homogeneity and significant results respectively. Results: Five RCTs with 7 interventions involving 2812 patients were included. The pooled odds ratio (OR) for MBQT in intention-to-treat (ITT) analysis was 1.25 (95% CI: 0.96–1.61), showing a non-significant trend. No heterogeneity was detected (I² = 0.0%). In the modified ITT (mITT) analysis (2 studies), MBQT showed higher eradication (OR: 1.70, 95% CI: 0.00–1042.90), but wide CI and high heterogeneity (I² = 70.7%) limited interpretation. All studies were included in the per-protocol (PP) analysis, which showed a statistically significant improvement with MBQT (OR: 1.67, 95% CI: 1.14–2.45) and low heterogeneity (I² = 5.2%), suggesting consistent results. Although not statistically significant, MBQT was associated with a slightly lower rate of adverse events compared to standard therapy (OR: 0.81, 95% CI: 0.59–1.12). I² = 50.6% showed moderate heterogeneity in safety outcomes. Discussion: the number of included RCTs was modest, with only five studies meeting eligibility criteria, and only two contributing to the modified intention-to-treat analysis. The risk-of-bias assessment showed variation in methodological quality across the included studies. Several studies exhibited high risk judgments in critical domains. particularly randomization, deviations from intervention, and selective reporting. Patients who completed the treatment benefited more from MBQT, which also had a comparable safety profile to conventional BQT regimens. In the treatment of H. pylori infection, MBQT may be considered a safe alternative for first-line treatment. Full article

Modifications in leaf architecture disrupt optical properties and internal light-scattering dynamics. Accurate modeling of leaf-scale light scattering is therefore essential not only for understanding how disease affects the availability of light for chlorophyll absorption, but also for evaluating its potential as an early optical marker for plant disease detection prior to visible symptom development. Conventional ray-tracing and radiative-transfer models rely on high-frequency approximations and thus fail to capture diffraction and coherent multiple-scattering effects when internal leaf structures are comparable to optical wavelengths. To overcome these limitations, we present a GPU-accelerated finite-difference time-domain (FDTD) framework for full-wave simulation of light propagation within plant leaves, using anatomically realistic dicot and monocot leaf cross-section geometries. Microscopic images acquired from publicly available sources were segmented into distinct tissue regions and assigned wavelength-dependent complex refractive indices to construct realistic electromagnetic models. The proposed FDTD framework successfully reproduced characteristic reflectance and transmittance spectra of healthy leaves across the visible and near-infrared (NIR) ranges. Quantitative agreement between the FDTD-computed spectral reflectance and transmittance and those predicted by the reference PROSPECT leaf optical model was evaluated using Lin’s concordance correlation coefficient. Higher concordance was observed for dicot leaves ($C_b=0.90$) than for monocot leaves ($C_b=0.79$), indicating a stronger agreement for anatomically complex dicot structures. Furthermore, simulations mimicking an early-stage fungal infection in a dicot leaf—modeled by the geometric introduction of melanized hyphae penetrating the cuticle and upper epidermis—revealed a pronounced reduction in visible green reflectance and a strong suppression of the NIR reflectance plateau. These trends are consistent with experimental observations reported in previous studies. Overall, this proof-of-concept study represents the first full-wave FDTD-based optical modeling of internal light scattering in plant leaves. The proposed framework enables direct electromagnetic analysis of pre- and post-penetration light-scattering dynamics during early fungal infection and establishes a foundation for exploiting leaf-scale light scattering as a next-generation, pre-symptomatic diagnostic indicator for plant fungal diseases. Full article

Listeria monocytogenes is a high-risk pathogenic bacterium associated with ready-to-eat foods and poses a potential threat to consumer health. This study aimed to investigate the prevalence, characterization and genetic diversity of L. monocytogenes in ready-to-eat raw salmon and trout products obtained from physical stores and online stores in China. Out of 150 samples analyzed, 23 (15.3%) were positive for L. monocytogenes. Among these positive samples, three (12%) were from Japanese restaurants, four (16%) from farmers markets, one (2.9%) from large supermarkets and fifteen (30%) from e-commerce platforms, and only one sample showed a contamination level exceeding 100 most probable number (MPN)/g. The isolates from positive samples demonstrated a concrete public health risk through several findings: twenty-three L. monocytogenes exhibited varying degrees of cytotoxicity, ranging from 7.6% to 71.8%. Compared with the reference strain ATCC 19115, five of these isolates were highly cytotoxic, a result that was validated by mouse survival rate experiment, which also confirmed their high virulence at tested dose. All isolates were resistant to cefuroxime sodium, ceftriaxone, cefepime and nalidixic acid, and 13% showed resistance to sulphamethoxazole-trimethoprim. Three serogroups were identified, with serogroup Ⅰ.1 (1/2a, 3a) being the most prevalent (65.2%). These isolates were grouped into eight sequence types, with ST8 (34.8%) and ST87 (30.4%) dominating. All isolates carried virulence genes associated with LIPI-1 andmultiple internalin genes (inlA, inlB, inlJ and inlK), confirming their potential pathogenicity. Additionally, the isolates harbored antimicrobial resistance genes lin and FosX. The five highly virulent isolates exhibited the highest genetic similarity to J2-031 (GCA_000438645.1) and C1-387 (GCA_000438605.1). The results provided valuable information for Chinese regulatory authorities to strengthen the risk monitoring of L. monocytogenes in ready-to-eat raw salmon and trout products. Full article

The Xinjiang Uygur Autonomous Region, also known as Xinjiang, China, is notable for its high diversity and abundance of lichens. The purpose of this study was to examine species diversity and the distribution patterns of saxicolous lichens, family Megasporaceae, which includes the genera Aspicilia, Circinaria and Lobothallia, in Xinjiang Province. Morphology, anatomy, chemical analysis and rDNA-ITS sequences for the species were employed for their identification. As a result, 34 crustose and strictly saxicolous species belonging to three genera were found, which included 22 species of the genus Aspicilia, two of which were new to Xinjiang (A. disjecta (Zahlbr.) J.C. Wei and A. pycnocarpa Q. Ren & Lin Liu), eight common species of Circinaria, as well as four species of Lobothallia, two of which (L. determinata (H. Magn.) T.B. Wheeler and L. pruinosa Kou & Q. Ren) are new provincial records. There was a unimodal pattern with respect to lichen species richness; all specimens of the Megasporaceae family were found between 1600 and 5100 m altitude. The 30 species were collected at altitudes between 2601 and 3100 m; only four species were recorded below 2150 m, and seven were found above 4600 m. As far as the type of rocks are concerned, 24 species were found on siliceous rocks and 10 species were found on calcareous rocks. The 24 lichen species contained seven different secondary metabolites; stictic acid, substictic acid and norstictic acid were more common, whereas aspicilin, constictic acid, lecanoric acid and connorstictic acid were found in only a few lichen species. Full article

Rapid and accurate phenotypic screening of rice germplasms is crucial for identifying potential sources of rice sheath blight resistance. However, visual and/or caliper-based estimations of coalescing, necrotic, diseased lesions of rice sheath blight (ShB)-infected plants are time-consuming, labor-intensive, and subject to human rater subjectivity. Here, we propose the use of RGB images and image processing techniques to quantify ShB disease progression in terms of lesion height and diseased area. To be specific, we developed a Pixel Color- and Coordinate-based K-Means Clustering (PCC-KMC) algorithm utilizing the Mahalanobis distance metric, aimed at accurately segmenting symptomatic and non-symptomatic regions within rice stem images. The performance of PCC-KMC, combined with manual classification of the segmented regions, was evaluated using Lin’s concordance correlation coefficient (${\rho }_c$) by comparing its results to visual measurements of ShB lesion height (cm) and to lesion/diseased area (cm₂) measured using ImageJ. Low bias (C_b) and high precision (r) were observed for absolute lesion height (C_b = 0.93, r = 0.94) and absolute symptomatic area (C_b = 0.98, r = 0.97) studies. Furthermore, to automatically classify the segmented regions produced by the PCC-KMC algorithm, we employed a convolutional neural network (CNN). Unlike conventional CNNs that require fixed-size image inputs, our CNN is designed to take the RGB histogram of each segmented region (a 1000 by 3 representation) as input and determine whether the region corresponds to ShB infection. This design effectively handles the arbitrary sizes and irregular shapes of segmentation regions generated by PCC-KMC. Our CNN was trained based on an 85%:15% composition for the training and testing dataset from a total of 168 ShB-infected stem sample images, recording 92% accuracy and 0.21 loss. PCC-KMC-CNN also showed high accuracy and precision for the absolute lesion height (C_b = 0.86, r = 0.90) and absolute diseased area (C_b = 0.99, r = 0.97) studies, indicating that PCC-KMC combined with automatic CNN-based classification performs very effectively. These results demonstrate that the potential of our methodology to serve as an alternative to the traditional visual-based ShB disease severity assessment and can be considered to be utilized for lab-scale, high-throughput phenotyping of rice ShB. Full article

Background: Ginseng oligopeptides (GOPs), small bioactive peptides with potent antioxidant capacity and high bioavailability, have shown promise in promoting healthy aging; however, their underlying molecular mechanisms remain largely unexplored. Methods: Using the model organism Caenorhabditis elegans (C. elegans), we comprehensively evaluated the anti-aging effects of GOPs on lifespan, locomotion, oxidative stress, and gene expression. Integrated phenotypic assays and transcriptomic analyses were conducted to elucidate GOP-mediated molecular mechanisms. The transgenic strain TJ356 (DAF-16::GFP) and the loss-of-function mutant CF1038 [daf-16(mu86)] were employed to functionally validate the role of the DAF-16/FOXO pathway. Results: GOP supplementation significantly extended median lifespan by approximately 11.5% and improved age-related locomotion decline in C. elegans. Transcriptomic profiling identified 1928 differentially expressed genes (DEGs) enriched in metabolic, antioxidant defense, and longevity-regulating pathways. GOPs upregulated key antioxidant and stress-response genes (gst-4, sod-5, mtl-1) and longevity-related regulators (daf-16, lin-31, Y51B9A.9, and daf-12), while downregulating ins-7, an insulin-like peptide. Moreover, GOPs enhanced cytochrome P450–related detoxification and vitamin-dependent (retinol, ascorbate, and riboflavin) metabolic pathways, establishing a multidimensional antioxidant defense network. Phenotypic validation confirmed that GOPs markedly reduced reactive oxygen species (ROS) levels and lipofuscin accumulation (p < 0.001). Notably, GOPs promoted DAF-16 nuclear translocation in TJ356 worms, whereas the lifespan-extending effects were abolished in CF1038 mutants, highlighting the essential role of DAF-16/FOXO in mediating GOP effects. Conclusions: GOPs delay aging in C. elegans by activating the DAF-16/FOXO signaling cascade and reinforcing antioxidant networks, thereby maintaining redox and metabolic homeostasis. These findings provide novel mechanistic evidence supporting GOPs as promising dietary antioxidants for promoting healthy aging through modulation of conserved redox and longevity pathways. Full article

Background/Objectives: Tinnitus treatment is often based on coping strategies, as, still, no causal treatment is available. Based on our “Erlangen model” of tinnitus development, we treated tinnitus patients with individualized, non-masking low-intensity noise (LIN) to reduce the tinnitus loudness. Methods: A total of 72 adult patients with a tinnitus severity index below IV (tinnitus health questionnaire, THQ), a hearing loss not exceeding 40 dB up to 6 kHz, and without experience in hearing aid (HA) usage were included in this study. Their audiograms, tinnitus parameters, THQ scores, and optimal LIN were determined individually. Patients were double-blinded, assigned to a treatment-only (TO) or placebo-and-treatment (PT) group. The TO patients received treatment for four weeks while the PT patients initially received a placebo (low-intensity white noise) stimulation for two weeks and, hereinafter, treatment for four weeks. Every two weeks, the measurements listed above were repeated. The long-term effects on THQ were assessed four weeks after the end of treatment. The data were analyzed by parametric and nonparametric statistics. Results: We identified a significant decrease in the THQ score already after two (η² = 0.14) and four weeks of treatment (η² = 0.43), that was still present four weeks after the end of treatment (η² = 0.15) in the TO group. Subjective individual patient reports hint at a possible complete suppression of the percept by LINTS. The PT group profited much less, as the initial placebo treatment seemed to counteract the LIN effects. Conclusions: Individually fitted LIN treatment seems to be able to suppress tinnitus, in some cases completely. The optimal fitting of the LIN is crucial for treatment success. Full article

Several factors, including semen quality, can influence fertilisation success. Poor semen parameters may necessitate more frequent inseminations or the removal of males with consistently low fertility. This study evaluated turkey ejaculates (n = 37) with good fertility (GF) and impaired fertility (IF). The analyses included sperm motility parameters (total motility—TMOT, progressive motility—PMOT, curvilinear velocity—VCL, straight-line velocity—VSL, average path velocity—VAP, linearity—LIN, straightness—STR, amplitude of lateral head displacement—ALH, and beat cross frequency—BCF), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), and nitric oxide (NO) production, as well as enzymatic and biochemical assays of semen, such as superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) activities, glutathione (GSH) content, malondialdehyde (MDA) levels, and zinc (Zn²⁺) concentration. In parallel, the proteomes of seminal plasma and spermatozoa were separated using SDS- and Tricine-PAGE, and selected proteins were identified by nano LC-MS/MS. Spermatozoa derived from IF ejaculates exhibited significantly reduced TMOT (p = 0.002), VCL (p = 0.028), and PMI (p = 0.000), accompanied by elevated STR (p = 0.000) and NO production (p = 0.044). In the seminal plasma of IF males, a significant decrease was noted in SOD (p = 0.000) and GPx (p = 0.001) activities, whereas CAT activity was markedly higher (p = 0.014). Seminal fluid from IF ejaculates was also characterised by increased GSH (p = 0.014) and MDA (p = 0.014) concentrations, accompanied by reduced Zn²⁺ content (p = 0.014). In contrast, IF spermatozoa exhibited elevated SOD activity (p = 0.001), but reduced GPx (p = 0.000) and CAT (p = 0.012) activities. Sperm cells from IF ejaculates also had lower GSH levels (p = 0.000), higher MDA concentrations (p = 0.000), and increased Zn²⁺ content (p = 0.018) compared with those from GF ejaculates. A proteomic analysis revealed differences in fertility-associated proteins: peroxiredoxin 6 (PRDX6) was detected exclusively in GF semen, whereas alpha-enolase (ENO1), fatty acid-binding protein (FABP7), cytoplasmic aspartate aminotransferase (GOT1), and L-lactate dehydrogenase B (LDHB) were detected only in IF semen. Overall, the results demonstrate that both semen parameters and proteome composition may potentially affect the fertilisation outcomes in turkeys. Full article

Adenomyosis (AM) is a hormonally responsive uterine disorder defined by ectopic endometrial tissue within the myometrium, causing pain, abnormal bleeding, and subfertility. Non-coding RNAs (ncRNAs)—including microRNAs (miRNAs), long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs)—are post-transcriptional regulators implicated also in uterine remodeling. We systematically reviewed original studies evaluating ncRNAs in AM using human samples, in vitro and animal models, or bioinformatic approaches. Data sources included PubMed and Google Scholar (inception up to 10 August 2025). Forty-one studies were included and synthesized across mechanistic, diagnostic, and translational domains. miRNAs (n = 31) were the most studied subclass, followed by lncRNAs (n = 10) and circRNAs (n = 5). Recurrent miRNAs such as miR-10b and miR-30c-5p (downregulated, inhibitory) and miR-145 (upregulated, promotive) regulate epithelial invasion, epithelial–mesenchymal transition, and cytoskeletal remodeling via PI3K–AKT/MAPK and Talin1 signaling. The let-7a/LIN28B axis governed estrogen-sensitive proliferation in the junctional zone, while miR-21 exhibited compartment-specific roles in decidualization and ectopic cell survival. Extracellular-vesicle (EV)-bornemiRNAs (e.g., miR-92a-3p, miR-25-3p, miR-4669) contributed to immune polarization and show early diagnostic potential. lncRNAs and circRNAs acted via chromatin modifiers and ceRNA networks. Most findings remain at the discovery stage. Convergent dysregulation was observed in key signaling pathways, including JAK–STAT, Wnt/β-catenin, and Hippo–YAP. ncRNAs regulate critical axes of invasion, proliferation, immune modulation, and hormonal response in AM. Targets with preliminary causal support—miR-10b/ZEB1, let-7a/LIN28B, and miR-145/Talin1—warrant further validation. Circulating miRNAs—especially in EVs—offer promise for non-invasive diagnosis. Full article

Propiconazole (PRO), a triazole fungicide, controls fungal diseases by disrupting ergosterol production in fungal cells. It is used in crops such as cereals and fruits. However, there are concerns regarding its potential to disrupt the endocrine system and cause reproductive toxicity. This study examined the effects of PRO on mouse testes, germ cells, and GC-1 spermatogonia. After eight weeks, PRO reduced testicular diameter and downregulated key germ cell genes (Sall4, Piwil, Nanos2, and Dazl). A histological examination revealed smaller seminiferous tubules and fewer SALL4+ cells. PRO also impaired steroidogenesis by downregulating genes (StAR, Cyp11a1, 3β-HSD1) and reducing sperm motility, with a decline in Velocity Straight Line (VSL), Linearity (LIN), Straightness (STR), and motile sperm. PRO caused dose-dependent cytotoxicity in GC-1 spermatogonia, decreased proliferation, and increased apoptosis, marked by cleaved caspase-3 and BAX. PRO also induced autophagy, as presented by elevated levels of autophagy-related genes (LC3 and ATG12) and proteins (ATG5 and LC3A/B). 3-Methyladenine (3-MA), an autophagy inhibitor, downregulates levels of autophagy- and apoptosis-related proteins when 3-MA and PRO are simultaneously treated in vitro. This suggests that both apoptosis and autophagy contribute to PRO-induced testicular cytotoxicity. This study is the first to detail that PRO affects sperm motility in mice and induces autophagy-mediated apoptosis in GC-1 spg. Full article

Listeria monocytogenes is a ubiquitous foodborne pathogen whose occurrence in food and food-processing environments raises public-health concerns, particularly when isolates carry antimicrobial-resistance determinants. Next-generation sequencing (NGS) is increasingly used to detect resistance genes and to predict phenotypic resistance. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA 2020) guidelines, PubMed, Web of Science, and Scopus were searched for original articles (2015–2024) that used second- and/or third-generation sequencing to characterize antibiotic resistance in L. monocytogenes from food and food-processing environments. After deduplication and screening, 58 studies were included from an initial 418 records. NGS reliably detected a set of recurrent resistance determinants across diverse sample types and geographies. The fosX locus (intrinsic fosfomycin-related marker) was effectively ubiquitous across studies, while acquired determinants were variably distributed: lin (35/58 studies, 60.34%), norB (33/58, 56.90%), and tetracycline genes overall in 20/58 (34.48%) with tetM as the most common (11/58, 18.97%). Reported concordance between the genotypes and phenotypes for acquired resistance was very high (>99% for most agents), with notable exceptions (e.g., ciprofloxacin and some fosfomycin cases). Common analysis pipelines and databases included ResFinder, CARD, BIGSdb-Lm, ABRicate, and ARIBA; most sequencing used Illumina short reads, with an increasing use of long-read or hybrid approaches. NGS is a powerful surveillance tool for detecting resistance determinants and for source-tracking, but its predictive value depends on integration with phenotypic testing, standardized reporting, and comprehensive, curated databases. Key gaps include inconsistent phenotype reporting, variable database coverage, and limited assessment of gene expression/regulatory effects. Full article

RBL1 (p107) is a member of the retinoblastoma (RB) family of pocket proteins involved in cell cycle regulation and E2F transcriptional repression. While its promoter contains conserved E2F motifs, the integrated regulation of RBL1 by upstream tumor suppressor pathways remains incompletely understood. Here, we investigate the p53-dependent transcriptional regulation of RBL1 and dissect the contribution of its tandem E2F binding sites to this mechanism. Luciferase assays in synchronized cells demonstrated that these two conserved E2F sites are required for cell cycle-dependent activation of the RBL1 promoter. Overexpression of p53 showed that p53 represses RBL1 promoter activity in an E2F site-dependent manner. Using HCT116 p21 knockout cells, we revealed that this p53-dependent repression is mediated by p21. Chromatin immunoprecipitation confirmed dynamic in vivo binding of E2F1–3 and E2F4, while DNA pull-down assays revealed specific in vitro recruitment of RB, p107, and E2F1-4 to the two E2F sites, along with weak binding of MuvB components. Additional experiments in RB^–/– and LIN37^–/– knockouts showed that RB/E2F repressing complex plays the main role in repressing the RBL1 promoter, while E2F4, p107, and p130 can support this effect to a lesser extent. Overall, our findings demonstrate that p53 controls RBL1 expression indirectly through the p21–RB–E2F pathway by utilizing two E2F binding sites within the RBL1 promoter. Full article