Search Results (1,279)

The AP2/ERF superfamily is a key class of transcription factors involved in plant responses to various stresses. As an ancient species, the olive tree (Olea europaea L.) exhibits considerable stress tolerance and wide adaptability. In this study, we identified 348 AP2/ERF genes in the cultivated olive variety ‘Arbequina’ at the whole-genome level. According to protein sequence alignments and phylogenetic analyses via the Maximum Likelihood method, these genes were classified into four major families: AP2, ERF/DREB, RAV, and Soloist. The ERF/DREB family was further divided into DREB and ERF subfamilies, each encompassing six groups (A1–A6 and B1–B6), with the ERF subfamily being the largest. Members of each group exhibited relatively consistent gene structures and domain/motif compositions of their encoded proteins; however, the distribution of cis-elements and expression patterns varied. Each AP2/ERF gene contained 12 light-responsive, three MeJA-responsive, three ABA-responsive, two anaerobic induction, and one MYB binding site on average. With the threshold of p value < 0.5, control TPM > 0, and |log₂(fold change)| > 0, 50 candidate genes were simultaneously up-regulated (30) or down-regulated (20) under four stress treatments (acid–aluminum, cold, disease, and wound), among which nine showed potential protein–protein interactions. This study provides a comprehensive genomic characterization of the AP2/ERF family in olive and identifies key candidate stress-responsive genes, establishing a foundation for future functional studies on the molecular mechanisms of stress adaptation in the olive tree. Full article

Torsional stress from DNA supercoiling is receiving renewed attention as a driving force for chromosome folding and the establishment of gene activity states. Transcription is a major source of DNA supercoiling, while topoisomerases relax supercoils and solve topological problems that arise during DNA replication, transcription, and chromosome segregation. Recent technological advancements have allowed for the mapping of how torsional stress distributes within the genome and distinguishing between occupancy of topoisomerases on chromatin and sites where they are catalytically engaged. Coupling these innovations to assessments of 3D chromosome conformation and nascent transcription at high resolution have provided a new understanding of the relationships between supercoiling and topoisomerase activity. Here, we summarize the insights obtained from these recent studies and discuss how the interplay between transcription, supercoiling, and topoisomerases shapes cellular gene activity states. Full article

Infectious bronchitis virus (IBV) causes a highly contagious disease in chickens. The prevalence of GVI-1 is increasing; however, the genomic characteristics and antigenic properties of this genotype strain remain insufficiently characterized. In this study, the genome characteristics and antigenic properties of a naturally attenuated CK/CH/SC/YC_GVI-1-DK/LMB20210104 (abbreviated as YC_GVI-1) strain were systematically analyzed. YC_GVI-1 occupies a distinct phylogenetic lineage and shares a similarity of 98.2%, the highest nucleotide sequence homology, with the reference strain CK/CH/FJ/202005 (accession number: MW791835.1). This strain was likely originated through a genetic recombination event between two major parental strains, CK/CH/FJ/202005 and CK/CH/GX/HX (accession number: PP817796.1). However, its S protein harbors ten unique amino acid substitutions, compared to the same protein in the other two virulent strains in the same genotype. AlphaFold3-based structural prediction reveals that one of these substitutions, methionine 485 to valine substitution, may induce a conformational change in the adjacent phenylalanine residue at position 431, resulting in a shift in the local secondary structure from β-sheet to random coil. Characterization of its antigenicity showed that this strain induces a strong humoral immune response, with neutralizing antibody titers of 2^6.40 against homologous strain YC_GVI-1 and 2^4.00 against heterologous strain JS96_GI-19. Furthermore, vaccination of chickens with this strain conferred complete protection (100%) against JS96_GI-19. The findings provide novel insights into the molecular evolution and antigenicity of YC_GVI-1, offering key information for improving IBV surveillance and vaccine development. Full article

The 2022 global mpox outbreak represented a turning point in the Monkeypox virus (MPXV) epidemiology, highlighting the incredible capability of this virus to adapt to different conditions, also in a non-endemic context. To investigate the genomic dynamics of MPXV 2022 strains, we performed whole-genome sequencing of 40 clinical samples from 16 Italian patients across multiple anatomical sites and timepoints between May and December 2022. Combining single-nucleotide analysis with detailed investigation of short tandem repeats (STRs), we explored inter- and intra-host viral dynamics. We identified 19 STR loci located near or within genes involved in immune modulation and virion morphogenesis. While most STRs remained stable across patients, a subset displayed locus- or matrix-specific variation. Among these, STR-VII—embedded within the coding sequence of OPG153, an envelope-associated protein implicated in viral attachment—showed a 12-nucleotide in-frame deletion, resulting in the loss of four aspartic acid residues (Δ4D variant). Structural modeling indicated that this deletion slightly alters a disordered acidic loop without affecting the global fold, potentially modulating surface charge and immune recognition. Integrating STR profiling into genomic surveillance may enhance resolution in outbreak reconstruction and reveal subtle adaptive processes underlying poxvirus–host interaction and immune escape. Full article

Single nucleotide variants (SNVs) are crucial in cancer occurrence and development. SNVs at the transcriptomic level generally come from genomic variants (g-tSNVs) and RNA editing (e-tSNVs). The types and quantities of e-tSNVs remain a subject of debate due to a relatively poor understanding of RNA editing processes. Herein, we developed TSCS (Transcript SNVs Classifier relying on complementary sequencings), a machine learning classifier that integrates short-read (MGI) and long-read (PacBio) RNA-seq data to accurately distinguish true transcript SNVs using stringent criteria. Applied to five colorectal cancer cell lines (HCT15, LoVo, SW480, SW620, and HCT116), TSCS demonstrated superior accuracy and sensitivity, outperforming established tools (GATK, BCFtools, Longshot, RED_ML). It increased the total detected transcript SNVs by 31.83% on average, with g-tSNVs and e-tSNVs exceeding conventional methods by >1-fold and >2-fold, respectively. TSCS achieved mean recall rates of 75.3% for g-tSNVs and 77.2% for e-tSNVs. Notably, for the first time, e-tSNVs were found in a relatively large proportion of total transcript SNVs in cancer cell lines, approximately 40%. Of the identified e-tSNVs, 80% were attributed to the known RNA editing, but the other e-tSNVs did not fall into any known category. Importantly, the e-tSNVs uniquely detected in this study showed distinct patterns in SNV types and genomic locations. Additionally, the transcript SNVs called by TSCS were partially confirmed using experimental approaches, such as Sanger sequencing, RNC-seq, and mass spectrometry. This study lays the foundation for surveying and appraising the cancer-related e-tSNVs. Full article

Food-borne diseases affect nearly 10% of the global population annually, with Salmonella being a major cause, particularly impacting children, the elderly, and populations in low- and middle-income countries. This study aimed to assess the prevalence, serotype distribution, antibiotic resistance profiles, and genetic determinants of resistance and virulence of Salmonella enterica in humans and poultry in the Nanoro health district. A community-based cross-sectional study involving humans and poultry was conducted in the Nanoro health district. Fresh stool samples (human and poultry cloacal/cecal) were collected, transported under sterile conditions, and processed within two hours using standard bacteriological methods. Phenotypic antibiotic resistance was determined by the Kirby–Bauer disk diffusion method, and whole-genome sequencing (Illumina) identified serotypes, resistance genes, and virulence factors. Logistic regression analyzed associations between Salmonella carriage and host or environmental factors. Salmonella enterica carriage was detected in 8.7% of humans and 7.2% of poultry. Human isolates showed 24% resistance to cephalosporins, while poultry isolates showed 36.8% resistance. Resistance genes, including fosA7, qnrB19, and a cryptic aminoglycoside resistance gene, and virulence genes encoding T3SS-1 and T3SS-2, were detected in both hosts. Logistic regression indicated that residence in Sitaon and Zimidin was associated with ~70% lower odds of carriage (aOR = 0.3), while individuals aged 11–20 and 51–60 years had 2.8-fold higher odds. Carriage was also 60% higher during the rainy season. These findings suggest possible cross-transmission of Salmonella between humans and poultry and the circulation of resistant, potentially virulent strains in the community. Seasonal and age-related variations highlight environmental and behavioral influences on asymptomatic carriage. Integrated One Health surveillance and targeted hygiene interventions are essential to reduce Salmonella transmission and antimicrobial resistance in rural settings. Full article

The radiosensitivity of cancer patients determines the efficacy of radiotherapy, and patients with low radiosensitivity cannot benefit from radiotherapy. Therefore, accurately predicting radiosensitivity before treatment is essential for personalized and precise radiotherapy. However, most existing studies rely solely on genomic and clinical features, neglecting the tumor microenvironmental information embedded in histopathological images, which limits prediction accuracy. To address this issue, we propose Resfusion, a deep multimodal fusion framework that integrates patient-level gene expression profiles, clinical records, and histopathological images for tumor radiosensitivity prediction. Specifically, the pre-trained large-scale pathology model is used as an image encoder to extract global representations from whole-slide pathological image. Radiosensitivity-related genes are selected using an autoencoder combined with univariate Cox regression, while clinically relevant variables are manually curated. The three modalities are first concatenated and then refined through a self-attention-based module, which captures inter-feature dependencies within the fused representation and highlights complementary information across modalities. The model was evaluated using five-fold cross-validation on two common tumor datasets suitable for radiotherapy: the Breast Invasive Carcinoma (BRCA) dataset (282 patients in total, with each fold partitioned into 226 training samples and 56 validation samples) and the Head and Neck Squamous Cell Carcinoma (HNSC) dataset (200 patients in total, with each fold partitioned into 161 training samples and 39 validation samples). The average AUC values obtained from the five-fold cross-validation reached 76.83% and 79.49%, respectively. Experimental results demonstrate that the Resfusion model significantly outperforms unimodal methods and existing multimodal fusion methods, verifying its effectiveness in predicting the radiosensitivity of tumor patients. Full article

DNA-protecting proteins (Dps) are crucial for safeguarding chromosomal DNA in starved cells during the stationary phase under stressful conditions. In previous research, the two Dps proteins in Deinococcus geothermalis, Dgeo_0257 (Dps3) and Dgeo_0281 (Dps1), were found to complement each other in protecting DNA from oxidative damage. This study investigates the physiological changes and transposition of insertion sequences (ISs) in a double-knockout (DK) mutant lacking both dps genes. Comparisons between the wild-type and mutant strains revealed significant phenotypic differences in viability under oxidative stress conditions induced by hydrogen peroxide and ferrous ions, particularly during the stationary phase. Notably, oxidative stress triggered the transposition of the IS families IS701 and IS5, with IS66 being transposed exclusively in the DK mutant into a gene encoding phytoene desaturase. Transcriptomic analysis using RNA-seq revealed substantial fold changes in gene expression across the genome. For example, the dgeo_1459–1460 gene cluster, which encodes a DUF421 domain-containing protein and a hypothetical protein, was highly upregulated under both oxidative and non-oxidative conditions. Interestingly, catalase, encoded by a single gene in D. geothermalis, was upregulated in the DK mutant during the stationary phase, with expression levels exceeding those observed in the single dps gene-deficient mutants. Conversely, a prominent downregulation of the Fur family regulator was detected. These findings highlight the growth phase-dependent physiological adaptation of the dps-DK mutant and reveal a novel IS transposition event of the ISBst12 group involving the IS66 family. Therefore, this study provides new observations into the influence of DNA-protective protein deficiency on oxidative stress responses and IS transposition in D. geothermalis, as well as the regulatory mechanisms of the catalase induction pathway, raising the need for further investigation into the role of OxyR. Full article

Background: The ACE Y215C mutation is a common, functionally damaging missense variant (~1.5% allele frequency) associated with reduced plasma ACE levels and increased Alzheimer’s disease (AD) risk. In CHO and HEK cell models, this mutation caused a ~3–6-fold decrease in ACE surface expression, soluble ACE levels, and ACE enzymatic activity compared to those of wild-type ACE. Methods: Circulating ACE levels and activity were measured in EDTA plasma obtained from 84 carriers of the ACE Y215C mutation using a set of mAbs to the ACE. The mAbs 5B3/1G12 binding ratio was revealed as a sensitive marker for the circulating Y215C ACE mutant. Whole-exome and whole-genome sequencing (WES/WGS) were performed to identify genetic variants potentially modifying circulating ACE levels. In parallel, published sequencing and proteomic data from 35,559 Icelanders participants were analyzed to identify genes influencing ACE shedding. Sequence comparison was performed between carriers with elevated and reduced ACE concentrations to identify the potential protective variants that may compensate for decreased ACE levels due to the Y215C mutation itself. Results: Most carriers of the Y215C ACE mutation demonstrated significantly decreased ACE levels (median is 62% of control ACE levels). However, substantial inter-individual variability was observed in plasma ACE activity among carriers. Comparative sequencing analysis revealed 9648 variants unique to individuals with elevated ACE, mapping to 5779 protein-coding genes and enriched for pathways related to intracellular and transmembrane transport. Conclusions: The presence of the damaging ACE mutation Y215C does not invariably result in low plasma ACE or, likely, elevated AD risk. Therefore, combined blood ACE phenotyping and whole-exome sequencing are recommended to more accurately assess ACE-related AD susceptibility in mutation carriers. Full article

Background/Objectives: Self-collection devices are more widely used than ever for detecting sexually transmitted infections and cervical cancer. Despite this, we still lack a clear understanding of how well these tools actually collect and release the necessary molecular samples. This study compared the in vitro uptake and release performance of commonly used self-sampling devices for total proteins, nucleic acids, and episomal human papillomavirus type 16 (HPV-16) DNA. Methods: An artificial cervicovaginal fluid composed of phosphate-buffered saline supplemented with serum and nucleic acid extracts was serially diluted 2-fold. Each dilution was applied for 5 min to the external surfaces of a vaginal veil (Vaginal Veil Collector V-Veil UP2^TM device), a flocked swab (FLOQSwabs^®), and a commercial vaginal tampon. Non-woven surgical tissue and plastic film served as controls. Total proteins and nucleic acids were quantified by spectrophotometry, and HPV-16 DNA by real-time quantitative PCR. Results: Recovery rates for proteins and nucleic acids were highest for the vaginal veil (81% and 91%), followed by the swab (66% and 70%) and non-woven tissue (44% and 47%). In contrast, the tampon and plastic film performed poorly, releasing less than 30% of proteins and negligible amounts of nucleic acids. Episomal HPV-16 DNA release was highest for the veil (89%), compared with the swab (57%), non-woven tissue (37%), tampon (4%), and plastic film (2%). Conclusions: The vaginal veil demonstrated superior uptake and release of proteins, nucleic acids, and HPV-16 DNA at physiological concentrations. Its non-absorbent structure allows high saturation with efficient release of genital components, including microbial genomes, whereas vaginal tampons retained these components, limiting analytical recovery. Full article

Phytosulfokine (PSK) is a tyrosine-sulfated pentapeptide found throughout the plant kingdom, playing key roles in plant growth, development, and responses to biotic and abiotic stresses. However, there is still a lack of a comprehensive analysis of the BnPSK gene family in Brassica napus. In this study, we conducted a genome-wide identification and characterized 19 BnPSK genes in oil seed plants, which are unevenly distributed across both sub-genomes (A and C). BnPSK proteins ranged from 77 to 99 amino acids (BnPSK3c and BnPSK3d) in length, all belonging to the PSK-α type and containing conserved PSK domains. Synteny analysis revealed that the expansion of the BnPSK gene family is primarily attributed to whole genome duplication, with homology to Arabidopsis thaliana PSK genes. A promoter region analysis identified cis-acting elements related to hormone and stress responses. An expression profile analysis showed that BnPSK genes are highly expressed in roots, leaves, petals, and pollens and are induced by both abiotic stresses and phytohormone application. Furthermore, RT-qPCR assay demonstrated that the expression levels of BnPSK4c, BnPSK5a, and BnPSK5b were significantly enhanced under drought stress (3~5-fold) both in plant roots and leaves following ABA application. Lastly, the application of ABA induced antioxidant activity including SOD, POD, CAT and APX (2~5-fold) and their corresponding genes (3~5-fold), and altered the ROS-signaling in rapeseed plants; also, strong evidence of mitigating drought stress was present. These findings establish a basis for further research into the role of the BnPSK gene family in oilseed plant tolerance against drought stress and underlying molecular mechanisms, offering valuable perspectives for developing novel peptides. Full article

Cetaceans are artiodactyls adapted to live in the marine environment, and this group includes whales, dolphins, and porpoises. Although mitochondrial nucleotide diversity has been reported separately for many cetacean groups, the proportion of deleterious mutations in these populations is unknown. Furthermore, a comparison of mitogenomic diversities across all cetaceans is also lacking. To investigate this, we conducted a comparative genomic analysis of 2244 mitochondrial genomes from 65 populations across 32 cetacean species. We observed a 78-fold variation in mitogenomic diversity among cetacean populations, suggesting a large difference in genetic diversity. We used the ratio of nonsynonymous-to-synonymous diversities (dN/dS) to measure the proportion of deleterious mutations in the mitochondrial exomes. The dN/dS ratio showed a 22-fold difference between the cetacean population. Based on genetic theories, the large differences observed in the two measures could be attributed to differences in the effective sizes of the cetacean populations. Typically, small populations have low heterozygosity and a high dN/dS ratio, and the reverse is true for large populations. This was further confirmed by the negative correlation observed between heterozygosity and dN/dS ratios of cetacean populations. While our analysis revealed similarities in mitogenomic diversity between the endangered and least-concern cetacean species, the dN/dS ratio of the former was found to be higher than that of the latter. The findings of this study are useful for identifying the relative magnitude of reductions in the population sizes of different cetacean species. This will help conservation management efforts prioritise the use of limited resources, time, and effort to protect the cetacean populations that need immediate attention. Full article

CRISPR-Cas9 systems have enabled unprecedented advances in genome engineering, particularly in developing treatments for human diseases, like cancer. Despite potential applications, limitations of Cas9 include its relatively large size and strict targeting requirements. Cas12j2, a variant ofCasΦ-2, shows promise for overcoming these limitations. However, its effectiveness in mammalian cells remains relatively unexplored. This study sought to develop an optimized CRISPR-Cas12j2 system for targeted knockout of the E6 oncogene in HPV-associated cancers. A combination of computational tools (ColabFold, CCTop, Cas-OFFinder, HADDOCK2.4, and Amber for Molecular Dynamics) was utilized to investigate the impact of engineered modifications on structural integrity and gRNA binding of Cas12j2 fusion constructs, in potential intracellular conditions. Cas12j2_F2, a Cas12j2 variant designed and evaluated in this study, behaves similarly to the wild-type Cas12j2 structure in terms of RMSD/RMSF profiles, compact Rg values, and minimal electrostatic perturbation. The computationally validated Cas12j2 variant was incorporated into a custom expression vector, co-expressing the engineered construct along with a dual gRNA for packaging into a viral vector for targeted knockout of HPV-associated cancers. This study provides a structural and computational foundation for the rational design of Cas12j2 fusion constructs with enhanced stability and functionality, supporting their potential application for precise genome editing in mammalian cells. Full article

Candidozyma auris (formerly Candida auris) is an emerging multidrug-resistant fungal pathogen with confirmed cases in over 30 countries. Although whole-genome sequencing (WGS) analysis defined distinct clades during characterization of underlying genetic mechanism behind multidrug resistance, Clade II remains under-evaluated. In this study, a three-level comparative genomic strategy (Global, Clade, Phenotype) was employed by integration of unbiased genome-wide comparative SNP screening (GATK v4.1.9.0), targeted BLAST profiling (BLAST+ v2.17.0), and in silico protein analysis (ColabFold v1.5.5; DynaMut2 v2.0) for systematic evaluation of mechanisms of antifungal resistance in thirty-nine Clade II C. auris clinical isolates and fourteen reference strains. Global and clade-level analyses confirmed that all the clinical isolates belong to Clade II, according to phylogenetic clustering and mating type locus (MTL) conservation. At the phenotype level, a distinct subclade of fluconazole-resistant mutants was identified to have a heterogenous network of mutations in seven key enzymes associated with cell membrane dynamics and the metabolic stress response. Among these, four core mutations (TAC1B, CAN2, NIC96, PMA1) were confirmed as functional drivers based on strict criteria during multitier in silico protein analysis: cross-species conservation, surface exposure, active site proximity, thermodynamic stability, and protein interface interaction. On the other hand, three high-level fluconazole-resistant clinical isolates (≥128 μg/mL) that lacked these functional drivers were subjected to comprehensive subtractive genomic profiling analysis. The absence of coding mutations in validated resistance drivers, yeast orthologs, and convergent variants suggests that there is an alternative novel non-coding or regulatory mechanism behind fluconazole resistance. These findings highlight Clade II’s evolutionary divergence into two distinct trajectories towards the development of a high level of fluconazole resistance: canonical protein alteration versus regulatory modulation. Full article

Genomic selection (GS) estimates the GEBV from genome-wide markers to reduce generation intervals and optimize germplasm selection, which is particularly advantageous for high-cost or late-expressed traits. While models like GBLUP are popular, they assume a polygenic architecture. In contrast, the Bayesian alphabet and machine learning (ML) can accommodate other types of genetic architectures. Given that no single model is universally optimal, stacking ensembles, which train a meta-model using predictions from diverse base learners, emerge as a compelling solution. However, the application of stacking in GS often overlooks non-additive effects. This study evaluated different stacking configurations for genomic prediction across 10 simulated traits, covering additive, dominance, and epistatic genetic architectures. A 5-fold cross-validation scheme was used to assess predictive ability and other evaluation metrics. The stacking approach demonstrated superior predictive ability in all scenarios. Gains were especially pronounced in complex architectures (100 QTLs, h² = 0.3), reaching an 83% increment over the best individual model (BayesA with dominance), and also in oligogenic scenarios with epistasis (10 QTLs, h² = 0.6), with a 27.59% gain. The success of stacking was attributed to two key strategies: base learner selection and the use of robust meta-learners (such as principal component or penalized regression) that effectively handled multicollinearity. Full article